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Smooth, rich and creamy milk chocolate with caramel and nutty overtones.
Ingredients: Sugar, cocoa butter, cocoa mass, whole milk powder, natural vanilla.
(cacao 44%, milk solids 19%)
Allergen information: See ingredients list in bold. May contain traces of nuts.
Suitable for vegetarians and gluten free.
Sustainable, ethically sourced cacao. | https://www.pureorigin.co.uk/product-page/venezuela-44-cacao-milk-chocolate-bar-85g |
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Published byZackery Basil Modified over 6 years ago
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Active Filters: concepts All input signals are composed of sinusoidal components of various frequencies, amplitudes and phases. If we are interested in a certain range of frequencies, we can design filters to eliminate frequency components outside the range Filters are usually categorized into four types: low-pass filter, high- pass filter, band-pass filter and band-reject filter. Low-pass filter passes components with frequencies from DC up to its cutoff frequency and rejects components above the cutoff frequency. Low-pass filter composed of OpAmp are called active filter (as opposed to lumped passive filter with resistor, capacitor and inductor) Active filters are desired to have the following characteristics: Contain few components Insensitive to component variation Not-too-hard-to-meet specifications on OpAmp Easy reconfiguration to support different requirements (like cutoff freq) Require a small spread of component values
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Applications of Analog Filters Analog filters can be found in almost every electronic circuit. Audio systems use them for pre-amplification, equalization, and tone control. In communication systems, filters are used for tuning in specific frequencies and eliminating others (for example, to filter out noise). Digital signal processing systems use filters to prevent the aliasing of out- of-band noise and interference.
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Butterworth low-pass filter Many low-pass filter are designed to have a Butterworth transfer function with magnitude response as follows: Graphs from Prentice Hall
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Low-pass filter: Sallen-Key Circuits Active low-pass Butterworth filter can be implemented by cascading modified Sallen-Key circuits. The Sallen-Key circuit itself is a 2 nd order filter. To obtain an nth order filter, n/2 SK circuits should be cascaded During design, capacitance can be selected first and then resistor values. As K increase from 0 to 3, the transfer function displays more and more peaking. It turns out that if K>3, then the circuit is not stable. Empirical values have been found for filters of different orders
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Example of a 4 th -order Lowpass filter by cascading two 2 nd -order SK filters
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Comparison of gain versus frequency for the stages of the fourth-order Butterworth low-pass filter.
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Butterworth high-pass filter By a change, the lowpass Butterworth transfer function can be transformed to a high-pass function.
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Butterworth high-pass filter: Sallen-Key By a change, the lowpass Butterworth transfer function can be transformed to a high-pass function. With real OpAmp, the Sallen-Key is not truly a high-pass filter, because the gain of the OpAmp eventually falls off. However, the frequencies at which the OpAmp gain is fairly high, the circuit behaves as a high-pass filter. Since the high-pass Sallen Key circuit is equivalent the same as the low-pass one, the empirical values for K would be still valid in this case also.
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Band-pass filter: Sallen-Key Circuits Graphs from Prentice Hall If we need to design a band-pass filter in which the lower cutoff frequency is much less than the upper cutoff frequency, we can cascade a low-pass filter with a high-pass filter. The below band-pass filter uses the first stage as a low-pass filter which passes frequency less than 10KHz and the second stage as a high-pass filter that passes only frequency above 100Hz. Thus, frequency components in-between is passed to the output.
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Figure 11.11 Bode plots of gain magnitude for the active filter of Example 11.2.
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A summary
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Application: Equalizer (EQ)
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All rights reserved. | http://slideplayer.com/slide/4173314/ |
For many hunters, the choice of a bow and arrow becomes a very personal one. Bow hunters have been in use for centuries and there are many stories about the usefulness of the weapon. Some stories talk about how the ancients developed the bow and how they were quite adept at using it in battle. Other stories talk about how the Chinese improved the bow to the point that it could be used in close quarter battle and to shoot arrows at incredible speeds. No matter how far apart the claims may be, one thing is certain; the Chinese were the first people to use a Mongolian bow.
The Mongolian bow was not an original creation of either China or Europe. It was actually created in Africa by a Berber named Muhammad al-Khan. While many believe he actually originated in Yemen, some say he lived in Egypt and headed an expedition to central Africa when he was captured and brought back with the Berbers to Arabize part of what was then known as Morocco. Regardless, of where he came from, the Mongolian bow is thought to have been derived from the Arab language. From there, the bow quickly spread throughout Africa, the Middle East, Asia and other areas of the world and has become a highly popular weapon to hunters everywhere.
While there are many different types of Mongolian bows, they all follow one basic design; that being, that of a curved spine on the top of the bow which allows it to curve while still remaining compact on the archer’s body. The bow then curves around the nock at the end of the string. This allows for an extremely fast release of the bow and increased accuracy. For this reason, many Mongolian bows are long enough that a person can hold them without any assistance.
There are a couple of other important factors in determining the rate of a Mongolian bow. They are the draw weight and the archer’s wrist strength. A weak wrist will affect the draw weight, since it determines the tension that is applied to the string. This then affects the speed and power of the bow. A Mongolian is much more than a hunting gun, since it is a tool to use in close range shooting.
When it comes to accuracy, the best way to test it is through a live animal. A hunter should aim to hit their target in the center of the eye, which is about a quarter of a millimeter above the eye. If the bow hunter doesn’t hit this mark, then the string is too weak or they are not using the right draw weight. Different animals will shoot differently, but the goal is still the same. A hunter must be able to strike their target dead on.
In order for a Mongolian bow to stay on target, the hunter must hold the bow steady. This isn’t always easy when the draw speed is fast. If the hunter pulls the bow, the limbs will move back and forth quickly. This will increase the amount of sideways movement, which will negatively affect accuracy. It is important to keep the Mongolian bow steady while in this state. The best way to do this is to make sure that the hunter keeps their arm straight, with the elbow resting on the ground.
The final factor to consider with a Mongolian bow is how accurately it shoots the arrows. The hunter shouldn’t rely just on the fire power alone, since speed can be deceiving. The best indicator of how accurate an arrow will fly is to take a few practice shots. Once a Mongolian bow user has learned their arrows, they should be able to take shots with any of their skills at an extremely fast rate, without having to worry about accuracy.
It is often said that practice is the best teacher. Just like learning to hunt, or anything else in life, if you are going to hunt with a bow you need to practice. A Mongolian bow hunter who doesn’t practice will have a difficult time in the long run, because he won’t be able to rely on their skill alone. A Mongolian bow user must be willing to put in the practice necessary to become an excellent bow hunter.
Other Interesting Articles:
What Are Draw Weights for Mongolian Bows? | https://silkroadmongolia.com/mongolian-bow-fire-rate-arrow-speed/ |
Q:
Function is continuous if graph is compact.
Let $X$ be a Hausdorff space and let $f:X\to \mathbb{R}$. If grapph of $f$ is compact we have to show that $f$ is continuous.
Since every closed subset of a Hausdorff space is closed, therefore grapph of $f$ is closed. WE know that if $f:X\to Y$ and $Y$ is compact, then graph of $f$ is clsed implies $f$ is continuous. But here $\mathbb R$ is not compact. Please help!
A:
Proof without nets:
Let $G$ be the graph of $f$.
Suppose $G$ is compact.
Let $A$ be closed in $\mathbb R$. We show $f^{-1} [A]$ is closed.
Note that $f^{-1}[A]=\pi_X [G\cap (X\times A)]$.
Since $G\cap (X\times A)$ is compact and the projection $\pi _X$ is continuous, $f^{-1}[A]$ is compact.
$X$ is Hausdorff, so every compact subset of $X$ is closed. DONE!
| |
PURPOSE: To provide a mating cage for small animals capable of mating male and female small animals semi-automatically under an artificial control, and breeding small animals efficiently and steadily.
CONSTITUTION: A small animal mating cage contains a main frame of cage 1 capable of accommodating female and male small animals, a partition board 2 formed in the main frame of cage and dividing the main frame of cage into two, a shaft portion 9 fitted as freely slidable to the partition board 2, and a gate part formed at a right place of the partition board and opened by movement of the shaft portion 9 allowing a free pass of small animals through it.
COPYRIGHT: (C)1995,JPO | |
Q:
Python regex to catch Verilog port names
I have some output port statements in verilog text file as shown below and I want to capture Just port names.
Example sample:
output Y;
output Y1,Y2,Y3;
output Z;
output Q, QN;
Expected output I want is a list with output port names
(Y)
(Y1, Y2, Y3)
(Z)
(Q,QN)
I tried below code but it doesn't give me the list quite I want.
outputPortListPattern=re.compile(r'^\s*(output)\s+(.*);',flags=re.M)
with open(fileA) as fh_in:
with open(fileB, "w") as fh_out:
for line in fh_in:
match=outputPortListPattern.match(line)
if match:
matchlist = match.group(2)
print("Output port list={}".format(matchlist))
A:
If it is a list of port names you want, then:
import re
text = """output Y;
output Y1,Y2,Y3;
output Z;
output Q, QN;"""
[re.split(r',\s*', m.group(1))
for m in re.finditer(r'^output\s+(([^,;]+)(\s*,([^,;]+))*);$', text, flags=re.M)
]
Prints:
[['Y'], ['Y1', 'Y2', 'Y3'], ['Z'], ['Q', 'QN']]
| |
Representation learning is concerned with training machine learning algorithms to learn useful representations, e.g. those that are interpretable, have latent features, or can be used for transfer learning.
ICLR 2019 • tensorflow/models •
We study the problem of representation learning in goal-conditioned hierarchical reinforcement learning. In such hierarchical structures, a higher-level controller solves tasks by iteratively communicating goals which a lower-level policy is trained to reach.
CONTINUOUS CONTROL HIERARCHICAL REINFORCEMENT LEARNING REPRESENTATION LEARNING
NeurIPS 2016 • tensorflow/models •
This paper describes InfoGAN, an information-theoretic extension to the Generative Adversarial Network that is able to learn disentangled representations in a completely unsupervised manner. InfoGAN is a generative adversarial network that also maximizes the mutual information between a small subset of the latent variables and the observation.
In recent years, supervised learning with convolutional networks (CNNs) has seen huge adoption in computer vision applications. Comparatively, unsupervised learning with CNNs has received less attention.
#6 best model for Conditional Image Generation on CIFAR-10
CONDITIONAL IMAGE GENERATION UNSUPERVISED REPRESENTATION LEARNING
EMNLP 2018 • tensorflow/models •
We therefore propose Cross-View Training (CVT), a semi-supervised learning algorithm that improves the representations of a Bi-LSTM sentence encoder using a mix of labeled and unlabeled data. On unlabeled examples, CVT teaches auxiliary prediction modules that see restricted views of the input (e.g., only part of a sentence) to match the predictions of the full model seeing the whole input.
SOTA for CCG Supertagging on CCGBank
CCG SUPERTAGGING DEPENDENCY PARSING MACHINE TRANSLATION MULTI-TASK LEARNING NAMED ENTITY RECOGNITION UNSUPERVISED REPRESENTATION LEARNING
In existing visual representation learning tasks, deep convolutional neural networks (CNNs) are often trained on images annotated with single tags, such as ImageNet. In this work, we propose to train CNNs from images annotated with multiple tags, to enhance the quality of visual representation of the trained CNN model.
IMAGE CLASSIFICATION OBJECT DETECTION REPRESENTATION LEARNING SEMANTIC SEGMENTATION TRANSFER LEARNING
ICCV 2017 • Tencent/tencent-ml-images •
The success of deep learning in vision can be attributed to: (a) models with high capacity; (b) increased computational power; and (c) availability of large-scale labeled data. What will happen if we increase the dataset size by 10x or 100x?
#9 best model for Image Classification on ImageNet
IMAGE CLASSIFICATION OBJECT DETECTION POSE ESTIMATION REPRESENTATION LEARNING SEMANTIC SEGMENTATION
Sentence vectors represent an appealing approach to meaning: learn an embedding that encompasses the meaning of a sentence in a single vector, that can be used for a variety of semantic tasks. Existing models for learning sentence embeddings either require extensive computational resources to train on large corpora, or are trained on costly, manually curated datasets of sentence relations.
NeurIPS 2017 • facebookresearch/poincare-embeddings •
Representation learning has become an invaluable approach for learning from symbolic data such as text and graphs. However, while complex symbolic datasets often exhibit a latent hierarchical structure, state-of-the-art methods typically learn embeddings in Euclidean vector spaces, which do not account for this property.
NiftyNet provides a modular deep-learning pipeline for a range of medical imaging applications including segmentation, regression, image generation and representation learning applications. NiftyNet enables researchers to rapidly develop and distribute deep learning solutions for segmentation, regression, image generation and representation learning applications, or extend the platform to new applications.
IMAGE GENERATION MEDICAL IMAGE GENERATION REPRESENTATION LEARNING
NeurIPS 2017 • williamleif/GraphSAGE •
Low-dimensional embeddings of nodes in large graphs have proved extremely useful in a variety of prediction tasks, from content recommendation to identifying protein functions. However, most existing approaches require that all nodes in the graph are present during training of the embeddings; these previous approaches are inherently transductive and do not naturally generalize to unseen nodes. | https://paperswithcode.com/task/representation-learning |
Until now, neither have been an issue. The other day, Landon was sitting on one of his little chairs that has toys attached to it, eating a teething biscuit. By the way, teething biscuits have to be one of THE messiest foods I’ve ever seen! Anyway, my mother-in-law was cleaning him up after he finished and Ella, sneakily and one by one, grabbed the toys which were now covered in biscuit crumbs. The toys were (<– notice how I said “were” not “are”) those round plastic rings, bright colored that stack on a cylinder and form a pyramid. Super flimsy and took no time at all for Ella to demolish while trying to eat every crumb she could find. I found the leftover plastic under our bed with some of the consumed plastic making it’s appearance this morning.
Welcome to our little slice of crazy town!
We have been so lucky to have some amazing photos taken over the last year and a half since finding out a little babe was on the way. Something I’ve been so bad about, in recent years, is displaying personal photos at home. This, to me, is such a down side to all the technology we use on a daily basis. Our photos are all contained on phones and computers and rarely make their way onto paper and into our homes. Why? One of my favorite things in other people’s homes is seeing all their personal photos, their family & friends, big events, love. I am resolving to have more photos printed and finish my home spaces with this personal décor detail.
I had this large wall space in our living room and went back and forth about what to put there. We did have a gallery wall in the house before, but it was all wedding photos and the location wasn’t a noticeable spot where people would stop to look at it. I asked Jeff if I should replace the current gallery wall or move it to the large wall in our living room. His response, “put it in the living room so we will actually see it everyday and enjoy it”. Yes! Too often, gallery walls are hidden on stairwells and in hallways where people don’t see them as often. Bring those photos to a prominent location!
Figure out where you are going to put your new gallery wall. Chances are, you already have a blank wall in your home that you struggle with filling or decorating. Next, think about how big you want the gallery to be. This will give you a measurement from top to bottom and side to side and help you know what size and how many frames you can fit in your space. For instance, my space was about 42″ x 51″. I had a pretty particular look I wanted, so I had already picked out the frames and therefore knew the exact measurements. I used an 11×14 frame that was matted for an 8×10 photo, plus the width of the actual frame, it left about 2″ between each photo if I did 3 across and 3 down. This part may take a little configuring, just be sure to measure the actual frame because depending on the thickness of the frame, it can add an additional 1-2″ to each photo.
Here’s a tip if you want a uniform look, use a frame with matting. This will help separate the photos from each other as well as “tone down” the photo so that each picture isn’t competing with the one next to it. I was going for a black & white theme and found these frames for an amazing price at Michael’s. This frame is normally $23, but Michael’s regularly has sales on their frames and they usually do it by brand. These were 60% off last week!!! While they are no longer on sale, they do have a few other brands on sale right now. If you find one you like that is full price, it’s worth it to just keep checking back for sales.
For the uniform grid look, you need to choose all vertical or all horizontal pics. I know I know, duh! But, I started picking out photos and then realized I was choosing both and had to go back and do it again! Another thing I did for uniformity was adjust the color of the photos before printing. The main reason I did this is because I used photos from all different photo shoots and 2 different photographers who have very different editing styles. Some of the photos were bright, vivid color and some were more muted. Plus, with different outfits and backdrops in each shoot, it just made for a lot of different looks going on. By adjusting the color, it helped make all the photos look more like they went together. I don’t have any fancy photo editing program, just the one that came loaded on my computer. All I did was edit the saturation of the photo to pull some of the color out. When I found a color level that I liked (I used -65 but it will probably be different on all computers/programs) I adjusted all the photos to the same level. Note: When you do this, make sure you “save a copy” of the photo, don’t save over the original, just in case you don’t like it, plus, you don’t want to loose the integrity of the original photo. Obviously this step isn’t necessary. I also did this because of the black & white look I was going for. I wanted to use my favorite photos but I didn’t want them completely black & white so this allowed me to get the look I was going for while still keeping a little color. As far as printing, if you are using professional photographs, chances are, your photographer will have a suggestion as to where to print them for the best quality. Our photographer uses Smugmug, but also suggests Shutterfly as a good place to print.
I’ll be honest, I was not super technical with this part. I hung the center photo where I wanted the center of the gallery to be. For the photos directly above, below and beside the center photo, I just measured out the height or width of the frame, plus an additional 2″ for the space between each photo. Because I was just sort of winging it, I did have to make some adjustments. Just step back and check after you hang each photo because it may look even up close but once you step away you’ll be able to tell for sure. If you want to be more technical about it, you can pick up a laser lever at the hardware store. This one from Lowe’s is $20, attaches to the wall and will easily get the job done. If you are using a laser lever, your laser line is going to be where you hammer the nails into the wall so be sure to adjust down a bit from where you want the top of the frame to be. | https://justlikeplayinghouse.com/2017/06/20/gallery-wall/ |
CROSS-REFERENCE TO RELATED APPLICATIONS
BACKGROUND
(i) Technical Field
(ii) Related Art
SUMMARY
DETAILED DESCRIPTION
Description of Apparatuses
Specific Example Processing
Others
This application is based on and claims priority under 35 USC 119 from Japanese Patent Application No. 2017-219434 filed Nov. 14, 2017.
The present invention relates to an information processing system, an information processing apparatus, and a non-transitory computer readable medium.
As a method for extracting a message related to content selected by a user, a method for extracting only one message connected to the content on a one-to-one correspondence basis has been known. However, if multiple users transmit messages related to the content, and if the one message only is extracted, it is difficult to recognize communication related to the content performed by the multiple users.
According to an aspect of the invention, there is provided an information processing system including a memory, a content display, an extraction unit, and a message display. The memory stores multiple pieces of content and multiple messages transmitted by multiple users. The content display displays the multiple pieces of content. The extraction unit extracts multiple messages from the memory if a user selects at least one piece of content of the multiple displayed pieces of content. The multiple extracted messages refer to the at least one piece of content or have the at least one piece of content attached to the multiple extracted messages. The message display displays the multiple messages extracted by the extraction unit.
Hereinafter, exemplary embodiments of the present invention will be described with reference to the attached drawings.
FIG. 1
1
is a view illustrating the overall configuration of an information processing system .
1
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1
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The information processing system is provided with a management server that is an example of an information processing apparatus. Further, the information processing system is provided with multiple terminal apparatuses operated by users. Each terminal apparatus is configured as a computer.
200
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300
400
Specifically, the terminal apparatus is configured as, for example, a smartphone, a tablet, or a personal computer (PC). The terminal apparatus communicates with the management server via a communication network such as the Internet.
FIG. 1
200
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Note that illustrates a case where three terminal apparatuses are provided, but the number of terminal apparatuses is not particularly limited.
FIG. 2
200
is a view illustrating the hardware configuration of each terminal apparatus .
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The terminal apparatus is configured as a computer and is provided with a CPU , a random access memory (RAM) , and a read only memory (ROM) . The terminal apparatus is also provided with a memory configured as a hard disk device, a flash memory, or the like.
200
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Further, the terminal apparatus is also provided with a communication interface (communication I/F) for communicating with external apparatuses.
200
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Further, the terminal apparatus is provided with a display functioning as part of a content display and a message display.
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The display includes, for example, a monitor using a touch panel system. The display receives an operation performed by a user and displays information for the user.
201
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A program to be run by the CPU may be provided to the terminal apparatus in a state of being stored in a computer readable recording medium such as a magnetic recording medium (such as a magnetic tape or a magnetic disk), an optical recording medium (such as an optical disk), a magneto-optical recording medium, or a semiconductor memory. The program to be run by the CPU may also be downloaded to the terminal apparatus by using a communication medium such as the Internet.
FIG. 3
FIG. 3
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is a view illustrating functional units implemented by the CPU or the like of the terminal apparatus . Note that illustrates only functional units that relate to a display process described later.
FIG. 3
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As illustrated in , the terminal apparatus includes an operation receiving unit , a display controller , and a transmitting/receiving unit .
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The operation receiving unit receives an operation performed on the terminal apparatus by an operator. Specifically, the operation receiving unit receives output from a mouse or the like operated by the user, output from the display if the display uses a touch panel system, or the like and thereby receives the operation performed on the terminal apparatus by the user (such as an operation for selecting a piece of content).
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The display controller functioning as part of the content display and the message display generates a signal for controlling the display and thereby performs display control of the display .
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The transmitting/receiving unit transmits and receives information to and from the management server .
FIG. 4
300
is a view illustrating the hardware configuration of the management server .
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The management server that is the example of the information processing apparatus is configured as a computer and is provided with a CPU , a RAM , and a ROM . The management server is also provided with a memory configured as a hard disk device or the like. Further, the management server is provided with a communication I/F for communicating with external apparatuses.
301
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A program to be run by the CPU may be provided to the management server in a state of being stored in a computer readable recording medium such as a magnetic recording medium (such as a magnetic tape or a magnetic disk), an optical recording medium (such as an optical disk), a magneto-optical recording medium, or a semiconductor memory. The program to be run by the CPU may also be downloaded to the management server by using a communication medium such as the Internet.
FIG. 5
FIG. 5
301
300
is a view illustrating functional units implemented by the CPU or the like of the management server . Note that illustrates only functional units that relate to the display process described later.
FIG. 5
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As illustrated in , the management server includes an information memory , a message extracting unit , a processing unit , and an output unit .
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The information memory that is an example of a memory stores therein multiple pieces of content and multiple messages transmitted from multiple users. The information memory stores each piece of content and at least one message related to the piece of content in connection with each other. More specifically, the information memory stores therein, for example, each piece of content and a message to which the piece of content is attached, in connection with each other.
312
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When the user selects a piece of content, the message extracting unit that is an example of an extraction unit extracts, from the information memory , multiple messages that refer to the piece of content or to which the piece of content is attached.
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To extract the messages referring to the piece of content, the message extracting unit first acquires, for example, information included in the piece of content, such as the title of the selected piece of content. The message extracting unit then extracts messages including the acquired information.
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The processing unit that is an example of a processing unit performs predetermined processing such as processing associated in advance with an operation performed by the operator by using the terminal apparatus .
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The output unit that is an example of an output unit outputs (transmits) information to the terminal apparatus or the like.
FIG. 6A
FIG. 1
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is a view illustrating an example of a display screen A (see ) of the display provided to each terminal apparatus .
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Note that display on the display is performed in response to an instruction from the management server . Specifically, the output unit of the management server outputs a display-related instruction to the terminal apparatus , and the display controller of the terminal apparatus performs the display control of the display in response to the instruction.
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FIG. 6A
Note that the display screen A described with reference to and succeeding drawings is used for managing projects executed by multiple users.
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The display screen A is shared by the multiple users. When one of the users accesses the management server by using the terminal apparatus , the terminal apparatus of the user displays the shared display screen A.
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FIG. 6A
The display screen A illustrated in is provided with multiple display areas. Specifically, four display areas that are a first display area , a second display area , a third display area , and a fourth display area are provided.
501
FIG. 6A
The first display area located in the upper left part of is a display area for a project list, and multiple ongoing projects are displayed in series.
501
502
504
In this exemplary embodiment, when a project is selected in the first display area , display in the other display areas that are the second display area to the fourth display area are changed. In these display areas, display is performed in accordance with the selected project.
502
The second display area is a display area for displaying project members who implement the project, and project members of a corresponding one of the projects are displayed therein.
502
In the second display area , the displayed project members vary depending on the project selected by the user.
503
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The third display area is a message display area, and multiple messages A transmitted by the project members are displayed therein. Also in the third display area , the displayed messages A vary depending on the project selected by the user.
In this exemplary embodiment, the term “message” denotes information intended by one of the project members to be notified to the other project members.
503
503
The messages A are displayed in the third display area in a time series.
503
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503
More specifically, in this example, the messages A are displayed in such a manner that the most recent message A is located at the lowermost position and less recent messages A are located at the upper positions.
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On the contrary, the messages A may be displayed in such a manner that the most recent message A is located at the uppermost position and less recent messages A may be located at the lower positions.
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Further, in the third display area , information regarding each of transmitters who transmits the corresponding message A (information such as a name identifying the transmitter) and information regarding the transmission date and time of the message A are displayed in association with the message A.
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503
Further, in this exemplary embodiment, as denoted by a reference sign P, each project member may add an image indicating a tag to a message A. This enables recognition of, for example, a message A with highly evaluated by the project members and a message A of interest to the project members.
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In a bottom portion of the third display area , a voice input button , a content attachment button , and a message transmission button are displayed.
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511
The messages A may also be input by the project members through voice input. To perform voice input, a project member selects (presses) the voice input button and utters to the microphone (not illustrated).
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503
A message A is thereby received and displayed in the third display area .
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313
FIG. 5
Note that as the message A displayed in the third display area , the message A itself input by the project member may be displayed. Alternatively, the message A may be translated by the processing unit (see ) and thereafter may be displayed.
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503
The content attachment button is a button operated by the project member when a piece of content (described later) is attached to a message A.
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503
In this exemplary embodiment, when the content attachment button is selected (pressed), a list of pieces of content (not illustrated) is displayed, and a piece of content selected from the list is attached to the message A.
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513
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The message transmission button is a button to be pressed by the project member when the content of a message A to be transmitted is determined. After the project member presses the message transmission button , the terminal apparatus operated by the project member transmits the message A to the management server .
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In this exemplary embodiment, the management server then receives the message A, and the message A is displayed on the terminal apparatuses of the other users.
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504
FIG. 3
The fourth display area is a content display area, and the display controller (see ) that functions as part of the content display displays images respectively depicting pieces of content related to a project (hereinafter, simply referred to as pieces of content ) in the fourth display area .
520
504
More specifically, the multiple pieces of content are displayed as thumbnail images in the fourth display area .
Note that the term “a piece of content” denotes a piece of information indicating a document, an image, voice, or the like.
It may be said that the piece of content is not only a text file but also a file related to a photo, a movie, or voice. It may also be said that the piece of content is not only a file but also a piece of information or the like indicating a web site (described later). In other words, it may be said that the piece of content includes a piece of information and that the piece of information handlable as a unit is a piece of content.
504
520
In the fourth display area in this exemplary embodiment, the pieces of content are visually displayed.
504
520
520
520
The fourth display area is designed to allow the arrangement of the pieces of content to be changed. For example, moving a piece of content to a different position by performing a drag-and-drop operation enables the arrangement of the pieces of content to be changed.
520
520
520
520
In addition, in this exemplary embodiment, superposing a piece of content on a different piece of content enables the multiple pieces of content to be arranged at a position of one piece of content .
FIG. 6A
520
504
520
520
504
Although illustrates the pieces of content in a file format in the fourth display area , information or the like indicating a web site may also be united to a pieces of content , and thus information not in the file format is also displayed as a piece of content in the fourth display area .
503
520
520
311
In this exemplary embodiment, if a message A is transmitted with a piece of content attached thereto, the piece of content is stored in the information memory .
520
311
503
520
503
More specifically, the piece of content is stored in the information memory together with the message A. In further explanation, the piece of content is stored in connection (association) with the message A.
520
503
311
520
311
503
520
In other words, in this exemplary embodiment, the piece of content and the message A are stored in the information memory , and in further explanation, the piece of content is stored in the information memory in connection (association) with the message A to which the piece of content is attached.
503
520
520
503
This exemplary embodiment thereby enables the message A associated with the piece of content to be recognized and, vice versa, enables the piece of content associated with the message A to be recognized.
311
520
503
520
In additional explanation, in this exemplary embodiment, the information memory that is the example of the memory stores the piece of content and the message A related to the piece of content in connection with each other.
503
520
520
504
503
503
Further, in this exemplary embodiment, when the message A is transmitted with the piece of content attached thereto, the piece of content is displayed in the fourth display area that is the content display area, and the message A is displayed in the third display area that is the message display area.
520
503
520
311
504
Note that the case where if the piece of content is attached to the message A, the piece of content is stored in the information memory and further displayed in the fourth display area has been described in this exemplary embodiment but is an example.
520
503
311
504
In another example, the piece of content referred to in the message A may be acquired by performing a search through the Internet or the like, stored in the information memory , and further displayed in the fourth display area .
FIG. 6A
560
504
560
520
In addition, as illustrated in , multiple selection images are displayed in a bottom portion of the fourth display area in this exemplary embodiment. One of the multiple selection images is selected by the user when the user performs an operation for processing a piece of content .
560
Specifically, the five selection images are displayed.
560
560
560
560
More specifically, in this exemplary embodiment, one of the selection images to be selected for translation (hereinafter, a translation selection image A) and one of the selection images to be selected for a review request (hereinafter, a review selection image B) are displayed.
560
560
560
560
560
560
Further, one of the selection images to be selected for an approval request (hereinafter, an approval selection image C), one of the selection images to be selected for optical character recognition (OCR) processing (hereinafter, an OCR selection image D), and one of the selection images to be selected for a suggestion request (hereinafter, a suggestion selection image E) are displayed.
520
504
560
560
520
In this exemplary embodiment, when a drag-and-drop operation is performed from a piece of content displayed in the fourth display area to one of the selection images , processing associated with the selection image is performed on the piece of content .
520
504
560
200
520
In other words, in this exemplary embodiment, when one of the pieces of content displayed in the fourth display area is moved to a position where the selection image is displayed (a predetermined position on the display screen A), processing associated with the position is performed on the piece of content .
560
520
560
520
520
If the translation selection image A is selected (if the piece of content is moved onto the translation selection image A), a document included in the piece of content is translated, and the piece of content including the document in a language different from the original language is generated.
313
520
FIG. 5
More specifically, the processing unit (see ) executes a translation process and generates the piece of content including the document in the language different from the original language.
560
520
560
If the review selection image B is selected (if the piece of content is moved onto the review selection image B), a notification indicating that a review is requested of at least one project member predetermined for a review is provided.
503
520
More specifically, information indicating the project member who gives a review is displayed in the third display area (the message display area) to indicate that a review is requested. The reviewer thereby gives a review of the piece of content .
560
520
560
520
If the approval selection image C is selected (if the piece of content is moved onto the approval selection image C), a notification indicating that approval is requested of at least one project member for approving the piece of content is provided.
520
503
520
520
Specifically, information indicating the project member who approves the piece of content is displayed in the third display area (the message display area) to indicate that approval is requested. The project member for approving the piece of content thereby approves or rejects the piece of content .
560
520
560
313
520
If the OCR selection image D is selected (if the piece of content is moved onto the OCR selection image D), the processing unit executes an OCR process of a document image in the piece of content .
560
520
560
313
520
311
212
200
503
If the suggestion selection image E is selected (if the piece of content is moved onto the suggestion selection image E), the processing unit searches for information related to the piece of content and thereby identifies the information (identifies the information by searching the Internet or the information memory ). The display controller of the terminal apparatus then displays the identified related information, for example, in the third display area (the message display area).
560
520
503
Note that the above-described five selection images may be used for not only the processing of the piece of content but also processing of different information such as a message A.
560
503
503
560
503
503
Specifically, for example, if the translation selection image A is selected for a message A (if the message A is moved onto the translation selection image A), the message A may be translated, and thereby a message A in a different language from the original language may be generated.
560
503
503
503
In addition, for example, if the suggestion selection image E is selected for the message A, related information related to the message A may be identified, and then the related information may be displayed in the third display area or the like.
503
Specifically, for example, if the message A indicates searching for a solution to an issue, for example, a solution to the issue may be suggested. Alternatively, for example, a person capable of addressing the issue may be identified and suggested.
FIG. 6A
570
504
570
504
570
504
In this exemplary embodiment, as illustrated in , an operation part is displayed in an upper right part of the fourth display area . The operation part is provided for receiving, from the project members, an operation for changing the display format of the fourth display area . In this exemplary embodiment, when the user operates the operation part , and thereby the display in the fourth display area is changed.
520
520
504
520
FIG. 6A
Specifically, in this exemplary embodiment, the pieces of content are displayed in a display format in which the pieces of content are displayed in a distributed state (the display format used for the fourth display area illustrated in ), a display format in which the pieces of content are displayed by forming a list, or by forming a lens view or the like.
200
501
504
200
FIG. 6A
FIG. 6A
Note that the four display areas shared by the multiple project members are displayed on the display screen A illustrated in , but in addition to the displays (in addition to the first display area to the fourth display area ) on the display screen A illustrated in , display areas for respective users (display areas that are not shared by the users but provided individually for the respective users) may be displayed.
520
520
520
520
In this case, if moving of a piece of content is performed between, for example, an display area specially provided for one of the users and a shared display area, a personal piece of content may be shared as a shared piece of content. On the contrary, if the shared piece of content is copied into the personal display area, each user may obtain the personal piece of content .
FIG. 6B
200
520
520
504
is a view illustrating the display screen A displayed after one of the pieces of content is selected from among the pieces of content displayed in the fourth display area .
FIG. 6B
200
520
520
504
More specifically, is a view illustrating the display screen A displayed in a case where Takagi who is one of the project members selects a piece of content from among the pieces of,content displayed in the fourth display area .
520
520
504
200
In this exemplary embodiment, if a project member (Takagi in this exemplary embodiment) selects at least one piece of content from among the multiple pieces of content displayed in the fourth display area , the display contents of the display screen A are changed.
520
503
520
200
Specifically, if the project member selects the piece of content , at least one message A related to the piece of content is displayed on the display screen A.
520
520
504
503
520
503
520
Specifically, in this exemplary embodiment, if at least one piece of content is selected from among the multiple pieces of content displayed in the fourth display area , multiple messages A referring to the selected piece of content or multiple messages A having the piece of content attached thereto are extracted.
520
312
311
503
520
503
520
More specifically, if the piece of content is selected, the message extracting unit extracts, from the information memory , the messages A referring to the selected piece of content or the messages A having the piece of content attached thereto.
212
503
503
6
FIG. 6B
Subsequently, the display controller that functions as the part of the message display displays the multiple extracted messages A in the third display area as denoted by a reference sign A in .
520
520
504
Note that a case where one of the project members (Takagi) selects a piece of content that is Rough sketch from among the multiple pieces of content displayed in the fourth display area is exemplified in this exemplary embodiment.
312
503
6
503
503
In this case, the message extracting unit extracts a message A that is a first message from Okubo (a message denoted by a reference sign B and also referred to as the first message A from Okubo) that is a message A referring to Rough sketch in this exemplary embodiment.
312
503
520
520
In additional explanation, in this case, the message extracting unit extracts the message A referring to the title of the piece of content that is information included in the piece of content .
312
503
520
520
520
Note that the message extracting unit extracts the message A on the basis of the title of the piece of content , but the basis is not limited to the title. For example, the content of the piece of content may be recognized, and thereby a message referring to the content (a message including the content of the piece of content ) may be extracted.
It may be said that a message referring to a piece of content is a message including information based on which the piece of content is identifiable.
503
6
Further, in this exemplary embodiment, a message A from Hasuike (a message denoted by a reference sign C) includes the phrase “Thank you for the rough sketch”.
312
503
Accordingly, the message extracting unit also extracts the message A from Hasuike that refers to Rough sketch in this exemplary embodiment.
314
300
503
503
503
312
Subsequently in this exemplary embodiment, the output unit of the management server outputs information regarding the two messages A (the first message A from Okubo and the message A from Hasuike that are extracted by the message extracting unit ).
200
503
312
503
The terminal apparatus thereby displays the two messages A extracted by the message extracting unit in the third display area .
312
503
503
6
200
503
Further, in this exemplary embodiment, the message extracting unit also extracts a message A that is the second message from Okubo (also referred to as the second message A from Okubo) denoted by a reference sign D, and the terminal apparatus also displays this message A.
312
503
503
503
503
6
503
Specifically, the message extracting unit regards the second message A from Okubo as a message A related to the above-described message A from Hasuike (the message A denoted by the reference sign C) and thus extracts the second message A from Okubo.
503
503
The extracted second message A from Okubo is also displayed in the third display area .
503
6
503
503
Note that the second message A from Okubo is provided with a mark X indicating a reply (provided with information indicating a reply), and thus the second message A from Okubo is a return message responding to the message A from Hasuike.
312
503
503
503
In this case, the message extracting unit regards the second message A from Okubo as a message related to the message A from Hasuike and thus extracts the second message A from Okubo.
503
503
In this exemplary embodiment, the second message A from Okubo is also displayed in the third display area .
Note that the return message responding to the message included in the two messages is the related message in this exemplary embodiment, but the related message is not limited to a return message.
312
If the message included in the two messages is a return message, a transmitted message that is the source of the return message is also a related message, and in this case, the message extracting unit also extracts the transmitted message.
312
More specifically, for example, if the first message from Okubo is a return message, a transmitted message that is the source of the return message is a related message, and the message extracting unit also extracts the transmitted message.
FIG. 6B
503
503
Further, in the display example illustrated in , the messages A transmitted by the project members other than the project member who selects the piece of content are displayed in the third display area .
520
503
503
Specifically, the project member who selects the piece of content is Takagi in this example, but the messages A transmitted by Okubo and Hasuike who are project members other than Takagi are displayed in the third display area .
FIG. 6B
520
520
520
504
520
In addition, as illustrated in in this exemplary embodiment, if the user selects the piece of content , any piece of content other than the selected piece of content is erased in the fourth display area . This makes the selected piece of content clearer.
520
520
Note that at least one more piece of content may be left in the display without erasing all of the other pieces of content .
520
520
In additional explanation, in this exemplary embodiment, one or more pieces of content included in the pieces of content other than the selected piece of content are erased.
520
520
520
Compared with a case where all of the other pieces of content are erased, the number of displayed pieces of content is reduced, and the selected piece of content is made clearer.
503
520
503
520
503
520
Although the case where the multiple messages A referring to the selected piece of content are extracted has been described, the extraction is not limited to this. If there are multiple messages A having the selected piece of content attached thereto, the multiple messages A having the piece of content attached thereto may be extracted.
More specifically, for example, if there is a message having a piece of content attached thereto, and further if there is another message having the piece of content attached thereto, the two messages may be extracted.
Also in this case, if there is a different message related to the extracted message like the case above, such as a return message or a transmitted message, also the different related message is preferably extracted.
520
503
520
503
In this exemplary embodiment, if the piece of content is selected, the multiple messages A related to the selected piece of content are displayed in this manner in the third display area .
503
503
503
Note that the multiple messages A are displayed in such a manner as to be arranged in a predetermined order. Specifically, the multiple messages A are displayed in a time series. This enables the project members to find an intended message A easily.
6
520
503
FIG. 6B
Further, as denoted by a reference sign Y in in this exemplary embodiment, an image representing the piece of content selected by the project member is also displayed in the third display area that is the message display area.
520
503
More specifically, a thumbnail image representing the piece of content selected by the project member is displayed also in the third display area .
520
503
520
503
6
Further, in this exemplary embodiment, the image representing the piece of content selected by the project member is arranged between the two messages A referring to the piece of content (the two messages A denoted by the reference sign A).
520
503
503
520
As described above, if the image representing the piece of content selected by the project member is displayed also in the third display area , it is possible to recognize, also in the third display area , which one of the pieces of content is selected by the project member.
520
503
503
520
520
504
In addition, if the selected piece of content is displayed also in the third display area that is the message display area as described above, the correspondence between the message A and the piece of content is displayed more clearly compared with a case where the selected piece of content is displayed only in the fourth display area .
FIG. 6B
520
503
200
Further, in this exemplary embodiment as illustrated in , the piece of content to be displayed and the multiple messages A to be displayed are displayed on the shared display screen A.
520
503
In addition, in this exemplary embodiment, the piece of content is displayed in one of display areas adjacent to each other, and the multiple messages A are displayed in the other display area.
503
504
520
504
503
503
Specifically, in this exemplary embodiment, the third display area and the fourth display area that are adjacent to each other are provided. The piece of content is displayed in the fourth display area that is one of the display areas, and the multiple messages A are displayed in the third display area that is the other display area.
520
503
520
503
200
520
503
The correspondence between the piece of content and the messages A is recognized more easily than in a case where the piece of content and the multiple messages A are displayed on the respective display screens A and a case where the piece of content and the multiple messages A are arranged in respective display areas spaced away from each other.
FIG. 7
200
504
is a view illustrating an example of the display screen A displayed after the display of the fourth display area is changed.
570
504
570
200
In this exemplary embodiment, the operation part for changing the display format of the fourth display area is provided as described above. When a project member operates the operation part , the display of the terminal apparatus of the project member is changed.
FIG. 7
200
200
1
3
0
520
520
7
illustrates the display screen A displayed after the display is changed. On the display screen A, pages P to P that are pages different from a cover P are displayed in at least one piece of content (in each of the two pieces of content denoted by a reference sign A).
520
200
520
FIG. 7
This enables the content of the pieces of content to be verified on the display screen A illustrated in without opening the pieces of content .
0
To display a page different from the cover P, a page satisfying a predetermined condition is displayed. Specifically, for example, a page highly frequently referred to by the user is displayed.
0
520
200
0
520
504
FIG. 7
Note that the pages different from the cover P are displayed in the at least one piece of content on the display screen A illustrated in , but the pages different from the cover P may be displayed in all of the pieces of content in the fourth display area .
200
520
509
520
520
200
520
520
520
FIG. 7
FIG. 7
Further, the display screen A illustrated in have the pieces of content of different sizes (frames of the respective pieces of content have different sizes). In additional explanation, the size of each piece of content is changed on the display screen A illustrated in in accordance with a predetermined condition. Specifically, the size of the piece of content is changed in accordance with the reference frequency or the like of the piece of content . More specifically, the higher the reference frequency, the larger the size of the piece of content .
520
Further, in this exemplary embodiment, a narrowing-down-operation receiving part (not illustrated) is displayed. The narrowing-down-operation receiving part is provided for receiving, from the project member, an operation for narrowing down the pieces of content .
520
In this exemplary embodiment, the pieces of content may be narrowed down (filtered) by operating the narrowing-down-operation receiving part or by inputting a keyword in an input box (not illustrated) provided in the narrowing-down-operation receiving part.
520
More specifically, the pieces of content may be narrowed down by using, as a key, information such as the number of reference times, a generation date and time, a creator, or a creation place.
504
FIG. 7
Note that the display of the fourth display area illustrated in may also be provided on a display of a copier installed in a convenience store or the like.
504
More specifically, a user is authenticated by using the copier installed in the convenience store or the like and is thereby identified. If the user is one of the above-described project members, the same display as in the fourth display area may be provided on the display of the copier.
520
520
This enables each project member to print any of the pieces of content also in the convenience store or the like. In addition, the display screen to be displayed at the time of printing is a display screen familiar to the project member, and thus the printing of the piece of content is facilitated.
FIG. 8
200
520
504
is a view illustrating a different display screen A displayed after a project member performs an operation for selecting a piece of content displayed in the fourth display area .
520
200
520
200
FIG. 6B
FIG. 8
Specifically, in this exemplary embodiment, performing the operation for selecting the piece of content leads to the display screen A illustrated in , but performing a different type of operation for selecting a piece of content leads to a change to the display screen A illustrated in .
520
200
200
FIG. 6B
FIG. 8
More specifically, in this exemplary embodiment, for example, double-clicking on the piece of content as the selection operation leads to the display screen A illustrated in , and performing an operation from a menu displayed by right-clicking leads to the display screen A illustrated in .
200
8
8
FIG. 8
On the display screen A illustrated in , an enlarged-content display area A and a message display area B are provided.
8
520
8
528
529
In the enlarged-content display area A, the selected piece of content is enlarged and displayed. The enlarged-content display area A is also provided with a version-information display area and a version change button .
8
503
520
In the message display area B, messages A related to the selected piece of content are displayed.
FIG. 6B
503
520
503
520
8
Specifically, like the description above (like the display illustrated in ), the messages A referring to the selected piece of content and a message A having the selected piece of content attached thereto are displayed also in the message display area B. Further, a related message such as a return message or a transmitted message is also displayed.
FIG. 6B
FIG. 8
In other words, the display in and the display in use different display formats but have the same display contents.
520
528
8
Version information regarding the piece of content thus enlarged and displayed is displayed in the version-information display area of the enlarged-content display area A.
520
311
529
528
FIG. 5
In this exemplary embodiment, a piece of content before version change is also stored in the information memory (see ), and if a different version is selected by operating the version change button , the display in the version-information display area is changed to the display indicating the different version.
529
520
8
In addition, if the different version is selected by operating the version change button , a piece of content corresponding to the selected different version is displayed in the enlarged-content display area A.
529
503
520
8
Moreover, if the different version is selected by operating the version change button , messages A related to the piece of content of the different version are displayed in the message display area B.
520
520
520
311
In this exemplary embodiment, after the piece of content is updated, the piece of content not updated and the piece of content thus updated are stored in the information memory .
520
503
520
520
520
311
More specifically, after the user updates the piece of content , and after, for example, a message A with the updated piece of content attached thereto is transmitted, the attached piece of content (the updated piece of content ) is stored in the information memory .
520
This also enables the piece of content of the different version to be verified in this exemplary embodiment as described above.
8
505
506
507
The message display area B is provided with display-content change buttons , a display-order change button , and a message display area .
505
505
505
505
As the display-content change buttons , a message display button A, a comment-in-page display button B, and a related-content display button C are provided.
505
503
520
8
8
When the user selects the message display button A, the messages A related to the piece of content displayed in the enlarged-content display area A are displayed in the message display area B.
505
505
8
505
505
When the user selects the comment-in-page display button B or the related-content display button C, the display of the message display area B is changed to different display. The display in a case of selecting the comment-in-page display button B or the related-content display button C will be described later.
506
503
507
In this exemplary embodiment, when the display-order change button is operated, the order of displaying the messages A in the message display area is changed.
506
506
503
Specifically, the display-order change button is a pull-down button. When the user operates the display-order change button , the messages A are displayed, for example, in the order from the greatest interest or in a time series.
6
FIG. 6A
Whether to be of great interest in the order from the greatest interest is determined on the basis of the number of tags as described above (see the reference sign P in ) or the number of affirmative evaluations such as “Like” (not illustrated).
503
FIG. 8
Note that the messages A may be narrowed down (filtered) by using a name or the like of a project member as a key, but this is not illustrated in .
503
503
507
Specifically, an input box may be provided. The messages A may be narrowed down on the basis of information regarding the project member input in the input box, and the messages A displayed in the message display area may be made different.
FIG. 9
FIG. 8
200
505
is a view illustrating the display screen A displayed after the user selects the comment-in-page display button B illustrated in .
505
520
8
8
If the user selects the comment-in-page display button B, a comment written for the piece of content displayed in the enlarged-content display area A is displayed in the message display area B.
FIG. 9
520
520
In the display example illustrated in , a comment for requesting replacing the illustration in the piece of content with the latest version illustration is displayed. Further, a comment for requesting making larger the font of characters in the piece of content is displayed.
FIG. 9
9
520
Further, in the display example illustrated in , an image (the image denoted by a reference sign A) is displayed, the image being for identifying characters font of which is to be made larger and being drawn by a user for the piece of content .
200
9
FIG. 9
Further, on the display screen A illustrated in , a read image obtained by scanning a document (a read image obtained by reading the document with a scanner (an image reader)) is displayed, as denoted by a reference sign B.
520
8
311
8
More specifically, this example exemplifies a case where a project member considers their document is related to the piece of content displayed in the enlarged-content display area A. This example further illustrates a case where the user performs a reading operation of the document and where the read image is stored in the information memory and is further displayed in the message display area B.
FIG. 10
200
505
is a view illustrating the display screen A displayed when the project member selects the related-content display button C.
505
520
520
8
8
When the project member selects the related-content display button C, pieces of content related to the piece of content displayed in the enlarged-content display area A are displayed in the message display area B.
520
520
520
8
520
520
520
8
Note that as an example of the related pieces of content , a piece of content transmitted together with the piece of content displayed in the enlarged-content display area A may be cited. In other words, multiple pieces of content are attached to one message, and at least one of the pieces of content that is different from the piece of content displayed in the enlarged-content display area A may be cited as an example.
520
520
503
503
520
8
In addition, as an example of the related pieces of content , a piece of content attached to a message A related to a message A to which the piece of content displayed in the enlarged-content display area A is attached may be cited.
520
503
520
8
Specifically, a piece of content attached to a return message responding to the message A to which the piece of content displayed in the enlarged-content display area A is attached may be cited as an example.
503
520
8
520
In addition, if the message A to which the piece of content displayed in the enlarged-content display area A is attached is a return message, a piece of content attached to a transmitted message that is the source of the return message may be cited as an example.
FIG. 11
200
520
is a view illustrating an example of the display screen A displayed when a project member writes a comment on the piece of content .
520
520
This exemplary embodiment is configured to allow a comment on any of pages constituting a piece of content while previewing the piece of content .
520
200
200
200
FIG. 6A or 7
FIG. 11
Specifically, in this exemplary embodiment, when a predetermined operation accompanying selection of a piece of content is performed by the project member on the display screen A illustrated in , the display screen A is changed to the display screen A illustrated in .
200
11
11
200
11
11
520
FIG. 11
FIG. 11
The display screen A illustrated in is provided with a preview area A in the left part of and a comment input area B in the right part of the display screen A. The preview area A and the comment input area B are respectively provided for previewing the piece of content and for inputting a comment.
520
520
This exemplary embodiment is configured to allow a comment on the piece of content without opening the piece of content .
520
520
520
11
11
11
520
More specifically, this exemplary embodiment is configured to refer to any page of the piece of content without launching dedicated software to open the piece of content (the piece of content is displayed in the preview area A). Further, in this exemplary embodiment, typing text in an input box C in the comment input area B enables commenting on the piece of content .
11
200
520
11
FIG. 11
Further, when a page change button D is operated on the display screen A illustrated in , a displayed page of the piece of content displayed in the preview area A is thereby changed.
11
311
520
11
FIG. 5
In this exemplary embodiment, the comment input by the user in the comment input area B is stored in the information memory (see ) in association with the piece of content displayed in the preview area A.
11
311
520
More specifically, the comment input by the project member in the comment input area B is stored in the information memory in association with the corresponding page of the piece of content .
505
11
507
FIG. 9
FIG. 11
FIG. 9
In this exemplary embodiment, if a project member selects the comment-in-page display button B as illustrated in , a comment input by a different project member in the comment input area B illustrated in is displayed in the message display area illustrated in .
FIG. 12
200
520
is a view illustrating the display screen A displayed after the comment on the piece of content is written.
520
503
FIG. 12
In this exemplary embodiment, when the comment on the piece of content is written, a notification indicating that the comment is written is displayed in the third display area as illustrated in .
520
503
In other words, in this exemplary embodiment, when a project member performs predetermined processing related to the piece of content , a notification indicating that the predetermined processing is performed is displayed in the third display area . The details of the processing performed by one of the project members is thereby notified to the other project members.
FIG. 12
520
12
520
The example illustrated in exemplifies a case where there is a comment on a piece of content denoted by a reference sign A. In this case, Update is displayed in association with the piece of content .
520
In other words, an indicator for identifying the piece of content processed by the project member is displayed.
503
12
12
12
520
12
520
12
520
12
In the third display area , the name of the project member who writes the comment (see a reference sign C), the date and time (see a reference sign D) when the comment is written, the content of the comment (see a reference sign E), and the piece of content commented (see a reference sign F) are displayed to indicate that the piece of content is commented (see a reference sign B). If the project member writes a note on the piece of content , the content of the note is also displayed (see a reference sign G).
520
520
504
503
In this exemplary embodiment, when a comment or a note is written in the piece of content , information for identifying the piece of content having the comment or the note is displayed in not only the fourth display area but also the third display area (the message display area).
520
520
504
This enables the project members to easily recognize the piece of content having the comment or the note compared with a case where the information for identifying the piece of content having the comment or the note is displayed only in the fourth display area .
FIG. 13
200
520
311
is a view illustrating the display screen A displayed when a new piece of content is stored in the information memory .
200
520
504
FIG. 6A or 7
In this exemplary embodiment, by performing a copy-and-paste operation or a drag-and-drop operation in a state where the display screen A illustrated in is displayed, a new piece of content may be pasted into (added to) the fourth display area .
520
520
13
504
520
311
504
FIG. 13
When a new piece of content (for example, a piece of content denoted by a reference sign A in ) is pasted into the fourth display area , the piece of content is stored in the information memory and is also displayed in the fourth display area .
520
520
13
520
13
503
520
13
FIG. 13
Further, when the new piece of content is pasted, the name of a project member who adds the piece of content (see a reference sign B) and the date and time when the piece of content is added (see a reference sign C) are displayed in the third display area (the message display area) to indicate that the new piece of content is added (see the reference sign A) as illustrated in .
520
503
13
Further, the added piece of content is displayed in the third display area (see a reference sign D).
520
520
504
503
In this exemplary embodiment, when the piece of content is added, information regarding the added piece of content is displayed in not only the fourth display area but also the third display area .
520
520
504
This enables the project members to easily recognize the newly added piece of content compared with a case where the information for identifying the added piece of content is displayed only in the fourth display area .
FIG. 14
200
520
is a view illustrating the display screen A displayed when approval of the piece of content is requested.
560
504
520
560
503
FIG. 14
In this exemplary embodiment, the approval selection image C is displayed in the fourth display area as described above. If the piece of content desired to be approved is moved onto the approval selection image C, a notification indicating that approval is requested is displayed in the third display area as illustrated in .
14
14
520
14
14
520
14
Specifically, the name of a project member who requests approval (see a reference sign B), the date and time when the approval is requested (see a reference sign C), and the name of a project member (an approver) requested to approve the piece of content (see a reference sign D) are displayed to indicate that approval is requested (see a reference sign A). Further, the piece of content to be approved is displayed (see a reference sign E).
520
14
520
14
14
Further, an Approve button that is a button to be selected when the piece of content is approved (see a reference sign F) and a Reject button that is a button selected when the piece of content is not approved (see a reference sign G) are displayed. Further, a comment input box for the approver (see a reference sign H) is displayed.
520
14
520
14
If the approver judges that the piece of content may be approved, the approver selects the Approve button (see the reference sign F). If the approver judges that the piece of content may not be approved, the approver selects the Reject button (see the reference sign G).
520
503
Thereafter, a result (result of whether the piece of content is approved or not approved) is displayed in the third display area (the message display area), but illustration of the result is omitted.
200
503
Also on the display screen A like the description above, an operation (for approval) performed by one of the project members is displayed in the third display area that is the shared display area, and the operation performed by the project member is notified to the other project members.
Information regarding the operation performed by the project member is thereby shared by all of the project members.
503
200
In this exemplary embodiment, simply operating the third display area enables the approver to perform the approval processing. In other words, the approver may perform the approval processing on the display screen A for managing the projects.
To perform the approval processing, for example, there is a method in which an e-mail or the like is separately transmitted to the approver to request approval, but it is laborious in this case. In addition, in this case, it is difficult for the other project members to share information regarding the approval processing.
200
In contrast, this exemplary embodiment enables the approval processing to be performed on the display screen A for managing the projects and thus facilitates the approval processing. In addition, the information regarding the approval processing is displayed on the shared screen and thus is shared by all of the project members.
200
Note that in this exemplary embodiment, the Approve button, the Reject button, and the comment input box are configured to be displayed on the display screen A of the approver only and not to allow approval of the project members other than the approver.
FIG. 15
200
560
is a view illustrating the display screen A displayed when the user selects the suggestion selection image E.
560
15
In this exemplary embodiment, only the suggestion selection image E may be selected. In this case, an image denoted by a reference sign A is displayed.
520
560
520
560
560
313
FIG. 15
FIG. 5
In additional explanation, the case where the piece of content and the suggestion selection image E are selected has been described (the case where the selected piece of content is moved onto the suggestion selection image E has been described), but illustrates a case where only the suggestion selection image E is selected. In this case, the processing unit (see ) performs processing and gives a suggestion.
560
313
300
503
311
520
311
Specifically, after the suggestion selection image E is selected, the processing unit of the management server analyzes information regarding the content of each message A stored in the information memory , the content of each piece of content stored in the information memory , the project members, and the like.
313
300
Further, the processing unit takes the analysis result into consideration and searches the management server and an external different server for information.
313
300
The processing unit identifies information to be suggested to the project members from among pieces of information stored in the management server and pieces of information stored in the different server or the like.
300
200
300
200
FIG. 15
The management server transmits the identified information to the terminal apparatus . Material suggested by the management server is thereby displayed on the terminal apparatus as illustrated in .
More specifically, for example, useful information and information regarding a helpful person for promoting a project are suggested.
Note that the suggested material is not limited to text information, a still image, and the like, and a moving image and a sound-related file may be suggested.
520
520
313
FIG. 5
Each piece of content may have an image added thereto, the image including information for identifying the piece of content (such as a code image). Note that, for example, the processing unit that is an example of an adding unit (see ) adds the image.
520
520
520
520
In this case, when the piece of content is printed, information for identifying the piece of content is added to a sheet having the piece of content printed thereon. More specifically, for example, a code image such as a barcode or two-dimensional barcode is formed, and thereby the information for identifying the piece of content is added thereto.
Thereafter, for example, when the sheet is read by scanning, the read image thereby includes the code image.
311
300
520
520
520
In this exemplary embodiment, if the read image is stored in the information memory (in the management server ), the code image in the read image is analyzed, and thereby the piece of content (the piece of content based on which the printing is performed) is identified. The piece of content and the read image are then associated with each other.
520
This processing facilitates association between, for example, a modification instruction or the like handwritten by a project member and the piece of content .
311
504
300
Note that the above-described read image may be stored in the information memory , for example, by performing a drag-and-drop operation or a copy-and-paste operation of the read image into the fourth display area . Alternatively, the read image may be directly transmitted from a scanner to the management server .
520
200
520
When multiple notes on the piece of content are displayed on the terminal apparatus , display of the multiple notes may be configured not to be simultaneously provided for one still image (a still image representing the piece of content ). Instead, for example, the notes may appear in order from the least recent note. In other words, the display may be changed in order, that is, an indicator for a note may be added to one another.
If multiple project members write a note, the color of the note may be made different for each project member.
The foregoing description of the exemplary embodiments of the present invention have been provided for the purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise forms disclosed. Obviously, many modifications and variations will be apparent to practitioners skilled in the art. The embodiments were chosen and described in order to best explain the principles of the invention and its practical applications, thereby enabling others skilled in the art to understand the invention for various embodiments and with the various modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the following claims and their equivalents.
BRIEF DESCRIPTION OF THE DRAWINGS
Exemplary embodiments of the present invention will be described in detail based on the following figures, wherein:
FIG. 1
is a view illustrating the overall configuration of an information processing system;
FIG. 2
is a view illustrating the hardware configuration of each of terminal apparatuses;
FIG. 3
is a view illustrating functional units implemented by a central processing unit (CPU) or the like of the terminal apparatus;
FIG. 4
is a view illustrating the hardware configuration of a management server;
FIG. 5
is a view illustrating the functional units implemented by a CPU or the like of the management server;
FIG. 6A
is a view illustrating an example display screen of a display provided to each terminal apparatus;
FIG. 6B
is a view illustrating the display screen displayed after one of pieces of content displayed in a fourth display area is selected;
FIG. 7
is a view illustrating an example display screen displayed after display in the fourth display area is changed;
FIG. 8
is a view illustrating a different display screen displayed after one of project members performs an operation for selecting a piece of content displayed in the fourth display area;
FIG. 9
is a view illustrating the display screen displayed after the user selects a comment-in-page display button;
FIG. 10
is a view illustrating the display screen displayed in a case where the project member selects a related-content display button;
FIG. 11
is a view illustrating an example display screen displayed when a project member writes a comment on the piece of content;
FIG. 12
is a view illustrating the display screen displayed after the comment on the piece of content is written;
FIG. 13
is a view illustrating the display screen displayed when a new piece of content is stored in an information memory;
FIG. 14
is a view illustrating the display screen displayed when approval of the piece of content is requested; and
FIG. 15
is a view illustrating the display screen displayed when a user selects a suggestion selection image. | |
How to Become a Content Writer – A Complete Guide 2022
Have you ever wanted to be a content writer, but didn’t know where to start? You will find the answers in this blog post. This article will you a complete guide on how to become a content writer and some useful tips and advice on content writing.
What is Content Writing?
Content writing is a job where you write text to provide information, education or entertainment to an audience. It could be for an article on a website or social media.
Content writing is creating an article that can be published on a blog or in a magazine or newspaper. Content writing involves research, proofreading, creative writing and editing.
What is content strategy?
Content strategy is a process that involves planning how content will be produced, how it will look, who it is targeted at, and how it will be distributed. By building your content writing standards, you get to be more creative with your writing as well as be more aware of the competition.
What Do Content Writers Do?
A content writer needs to be able to research topics and write about them in a way that is interesting and informative for the reader.
They must also have strong writing skills, especially spelling, grammar and the ability to describe things in detail. Content writers will also need to be able to take feedback on their work and be able to edit it, if necessary, to make it perfect.
Skills Required to Become a Content Writer
As a content writer, you need the ability to write in a way that will engage and interest people and make them want to read your work. Here is a list of skills that are required to become a content writer.
- Excellent Communication
- Good Writing Skills
- Creativity
- Excellent Proofreading Skills
- Strong Technical Knowledge
- Good Research Skills
- Time Management and Organizational Skills
- The Ability to Write for Different Genres (Relevant Experience is Required)
- SEO Knowledge
- Proper Keyword Research
Becoming a content writer is not an easy task. To be a successful content writer means that you would need to be able to write effectively, quickly and use the right vocabulary. You must also be aware of the changing trends and topics in your chosen field.
11 Effective Tips to become a good content writer
There are many qualities a good content writer should have if they want to get better at their job. Here I will give you 11 effective tips that will help you become a good content writer.
1. Get a Grip on Grammar
The most important thing that a content writer has to have is good writing skills. I agree that some can write well but still have bad grammar. If you don’t have good grammar then your chances of being accepted as a professional content writer are less than zero.
One of the easiest ways to improve your writing skills is to use speech recognition and editing software such as Grammarly or Google Docs for editing as and when you write. But never use bad grammar intentionally.
2. Write every day
Writing is a skill and like most skills, it will improve with practice. One of the best ways to improve your writing skills is to write a little bit every day. It may be two or three paragraphs but it should still be enough for you to proofread and edit your work. Build a habit of writing every day and keep on doing it.
3. Develop your voice
A good content writer will be able to develop their voice. This is something that comes with time. After writing several articles and getting feedback from your audience, you will be able to develop your style of writing and use it in the articles you write. Just be yourself and don’t try to imitate any other writer who is good at writing.
4. Observe the market
It is important to know what the market wants. To get a better understanding of what good content writing is, and what you should avoid, read and analyze content from content writers who are already making an impact in your chosen niche. Try to follow their style and even write a few articles based on their style.
5. Study the audience
The more you know about the audience you are writing for, the better. Take some time to study the audience before you begin writing. This will give you a better idea of where your article is heading and help you to write something that they want. It is a good idea to get some feedback from them before you finish the article.
6. Network with writers in your field
As a content writer, you should be talking to other writers and getting their feedback on your work. Show your work to other writers and get their feedback.
When you get the feedback, take the advice and improve your writing skills. Join community forums, e-mail lists and other professional networks.
7. Read, read & read
Reading is one of the most important things a content writer can do. It gives you examples of how to use words and phrases, it helps you find which words work best and it gives you a better understanding of what your audience likes to read.
You should make sure you read as much as possible. You can read for just 10 minutes every day or an hour every week, whatever works for you but don’t stop reading from improving your content writing skills.
8. Practice writing
To become a content writer you have to start somewhere. You have to start by reading and writing content on your own. I’m not saying that you should copy someone else’s content.
Don’t do that, it’s against the law. But the idea is to read a lot of articles and write even more on your own. Keep practicing until you become a pro.
9. Organize your thoughts and ideas
Organizing your thoughts and ideas before writing is something that every content writer should do. Organizing your ideas will help you plan how to write your content in the best way possible.
For example, if you organize your ideas properly you can make a clear plan of what the main sentences in your article should be like.
I don’t know if you have experience organizing your thoughts but it’s a difficult task. It’s even more difficult when you have to write a complete article on how to become a content writer. But it’s not impossible. You just have to organize your thoughts before you start writing.
10. Have a Passion
Content writing is a job that requires passion. You have to be passionate about what you are writing about. Think about it, if you have nothing to write about then it will be very difficult for you to write good quality content.
So make sure that your content writing job means a lot to you and there are no feelings of guilt or regret when you look at what you’ve written. To become a content writer you need to be more passionate about your work.
11. Never Give Up
You have to work hard and constantly practice writing every day. You have to get good at this because this is the very profession that you want to be in. Don’t give up because you can’t do it on the first try. Keep building your writing skill until you become the best in your field.
Bonus Tips:)
1. Use short sentences
2. Use bullet points
3. Ask questions
4. Read other content for inspiration
5. Be confident in your writing abilities
6. Don’t copy-paste
I hope these tips will help you to become a content writer. Keep practicing until you become a pro content writer.
How to Get Content Writing Jobs
The good thing about content writing is it’s a highly sought-after skill and there are job openings for people who have a good understanding of the industry. Here are some ways you can find jobs as a content writer:
1. Start Freelance Writing
Freelance writing is another option to help you get started. Freelancing is a quick and easy way to make money online.
You can take up freelance content writing jobs through online portals like Upwork, Peopleperhour, etc. You can also check out my list of 15 Best Website For Freelance Writers. This list will help you to find high-paying, freelance content writing jobs.
2. Start your Blog
If you want to start a blog, you can get online content writing jobs by writing articles on your blog. You may want to start with a niche blog that has the potential to generate high traffic.
Blogs are good platforms for beginners because they will help them learn more about blogging best practices, create a beautiful and professional-looking blog site and get high-paying content writing jobs. You will earn from ads and affiliate links from sites like Google AdSense and other affiliate programs.
3. Use Social media
If you’re good with social media, then it would be easy for you to find content writing jobs on social media. You can write for companies who offer their products and services through social media platforms like Facebook, Twitter, Instagram and LinkedIn.
With people’s need to stay connected, employers don’t mind paying a lot of money for content writing jobs on social media platforms.
You can also join Facebook and WhatsApp groups where you can post your content writing job and find many people who are interested in freelancing on those topics.
4. Write for the Newspaper or magazine
If you are looking to get into a stable, full-time job as a content writer, then you can start by writing for local newspapers or magazines. You can move up to bigger magazines and newspapers.
The key to getting content writing jobs in these organizations is to start from a local newspaper and then try your hand at other magazines and newspapers that are national or international.
5. Join Forum discussions
There are many places on the internet where people discuss issues that interest them. Some of these discussions can result in writing articles for online magazines or newspapers.
You may want to join discussions on a particular topic and let the other members know you are interested in writing articles for them. They will contact you if they need a writer.
Best content writing niches for beginners
There are three main types of content writing niches that can be best for beginners. Here are some niches/ideas you can start with.
1. Blog posts
Blog posts can be published on a blog, social media, as an article for a newspaper or magazine. By publishing a blog post, you are building a relationship with your audience, which is vital for the success of your business. Blog posts can be in any format such as an interview, story or white paper.
2. Ebooks and whitepapers
Ebooks and whitepapers are useful sources of information on a specific topic that can be downloaded by anyone. There is a lot of competition for this type of content, so it’s important to come up with a unique angle that will set your ebook or whitepaper apart from the rest.
3. Consumer research reports
This report comes in 3 or 4 different types: opinion polls, analytics reports and data analysis reports. These reports are submitted to different clients to show the results and the benefits of whatever it is they are testing.
4. Copywriting
This is a form of advertising used to promote a product or service. It involves writing content for websites, telemarketing and TV advertisements.
You will find that e-commerce is a large part of the copywriting niche. If you get into the e-commerce niche, this means that your job will be to write product descriptions for businesses selling products online such as Amazon.
5. Product Reviews
Product reviews are the best way to build trust and credibility with your audience. You will find that this niche is a good place to start as you will be able to get a lot of experience without too much competition.
However, it’s essential to research the best ways of writing your reviews. You can do it in different ways such as blog posts or articles for an e-zine.
Best Content Writing Tools
Content writing is all about making your readers or customers interested in the subject. You will have to have the best tools and resources at hand to make your article interesting, engaging, and long-lasting. Here are some best tools you can use as a content writer.
1. Grammarly
Grammarly is a grammar checker which analyzes and corrects your text to make it grammatically correct. It is a free web-based proofreader for editing text.
You can check your text for grammatical errors, spelling errors, and punctuation mistakes. It is a tool that every content writer should use at least once when they write an article.
2. Hemingway App
Hemingway app is a grammar checker and style checker which highlights the readability levels in your text. The readability level indicates how easy or difficult it is to read the text written. It helps you to communicate effectively by improving your sentence structure, word choice, and punctuation.
3. ProWritingAid
ProWritingAid is a free writing app for writing articles. It will help you make sure that you have a complete and perfect article. The app audits your already written article as well as lets you review your previous articles. The app helps you to become a better writer.
4. Thesaurus
Thesaurus is a vocabulary database of synonyms, antonyms, and related words. You can use this tool to make your writing better by using synonyms and words with the same meaning. It helps to avoid repetition of the same words in your article.
5. HubSpot’s Blog Ideas Generator
HubSpot’s Blog Ideas Generator will ask you a few simple questions, then it will provide you with blog ideas based on your answers. If you want to write about marketing, sales, blogging, and inbound marketing then this is the best option for your article.
6. Surfer SEO
If you want to know how to rank higher in search engines then this is the tool for you. Surfer SEO is a free online SEO audit tool that helps you in optimizing your website for Google.
You can search for keywords, check rankings, compare your rankings with competitors, and much more with this tool. It will help you write better content by understanding the content that works best.
7. Copyscape
Copyscape is a plagiarism checker which will help you to search for duplicate content. It is a good tool to check if your content has been lifted or copied. You can use this tool to check if the content on your website or blog has been stolen and published by someone else on their website.
There are also other tools like Google Docs and MS Office that can be used in content writing. You can make your tool kit according to your needs and requirements.
How much you can earn as a content writer?
The salary ranges from a few hundred to thousands of dollars. It depends on the nature of the requirement and experience. The average salary of a content writer is 15$.
But you can make more money as a content writer if you make a career in it. You can earn $100 to $700 per month. But this is something that you have to work hard for.
Mostly Asked Questions
How to start content writing?
You first need to join a social media platform such as LinkedIn and start networking with content writers and editors. You can also scout for jobs on sites like Upwork and Freelancer and learn the ropes by working there.
What is the qualification for a content writer?
You don’t need any specific qualifications to become a content writer. But, you must be a good writer. The only thing that matters is that you have the passion to write. Your writing skills are more important than your qualifications.
How to become a content writer with no experience?
You can become a content writer with no experience. The best way is to start networking and learn the ropes by working as a content writer.
You can also write articles for your local newspaper or magazine and get paid. You can join Facebook groups to learn more about content writing.
You can get experience by joining sites like Upwork or Fiverr to write content for other people. These are freelance writing sites where a client asks you to write anything like an article, blog post, content, product description, etc.
What platforms can I use for writing?
There are many online platforms for content writing. Some popular ones are LinkedIn, Upwork, Fiverr, Blogger, Medium, Docstoc and Hubspot.
How to improve your content writing skills?
There are several things you can do to improve your content writing skills. You can read articles on the topics that you want to write about, watch videos on YouTube and read blogs.
Sometimes, you get to experience different things and get a better perspective of what goes on outside when you participate in social media groups. That will help you write better content.
Final Words
In conclusion, content writing is a fun career. It will give you the privilege of meeting amazing, inspiring people as well as getting to do all the research and writing you can think of.
To become a content writer, you will need practice, research and a lot of patience. I hope this post helps you reach your goal of becoming a content writer. If you have any questions, feel free to ask me in the comment section. | https://beginnersjob.com/how-to-become-a-content-writer/ |
This 4-Day Workout split for optimal bulking and muscle mass takes into consideration that you will be consuming a high amount of caloriesper day (10-15,000). With an 8-12 inch waist and an average bodyweight of 160 lbs, you should be able to add another 20 lbs of muscle while doing the workout, not including the workout itself. It is important to know that when you have a diet that is restrictive then you will be at risk of wasting calories and your muscles will suffer from being depleted from eating these restrictive fast foods. It will be more appropriate to focus on a diet that is lower in calories, that does not dictate your portion size, and has a good calorie count of approximately 1,600 instead of 10,000, bulk up xl. What to eat for these 4 days: Breakfast: 4 breakfast cereals (I like PowerBar, Kashi and Smart Start), 2 eggs, 1 banana, 2 tbsp of peanut butter and about 100 grams of fat Lunch: 4 avocado, 2 whole wheat crackers, 8 ounces of salad and 5 cups water/pepsi/milk/coffee Dinner: 4 slices of toast, 10 oz. of steak (beef is great because it cuts down on any calories), 2 cups of rice, 2 cups of soup and about 150 grams of fat How to do the workout: The workout split for this workout is as follows: Monday: Legs Bout: 15 – 15 – 10:30 AM work set 15 – 15 – 10:30 AM work set Tuesday: Abs Bout: 20 – 15 – 10:30 AM work set 20 – 15 – 10:30 AM work set Wednesday: Back Bout: 20 – 15 – 10:30 AM work set 20 – 15 – 10:30 AM work set Thursday: Chest, Traps, Arms and Shoulders Bout: 20 – 15 – 10:30 AM work set 20 – 15 – 10:30 AM work set Friday: Rest What to eat for these 4 days: Breakfast: 4 ounces fruit, 2 eggs, about 70 grams of fat and 15 grams of protein Dinner: 4 slices of cheese, 8 ounces of salad with some tomatoes, 3 cups of meat soup, 3 cups of rice Do you want to add more muscle and lose fat on your next diet? You will be able to get leaner and lean faster, bulking 5x5 workout.
Bulking mean
Bulking steroids are to be used during bulking cycles when bodybuilders are looking to gain weightmore quickly after training. The process of taking steroids results in a change in hormonal levels, as well as an increase in muscle size. These steroids also cause the growth of muscle cells in the bulking cycle, bulking to gain weight. The effects of certain steroids are also visible during a natural workout, bulking workout plan no equipment. For example, testosterone causes changes in muscle growth, while cortisol causes muscle growth, bulk powders customer service. A combination of steroid usage can be a factor behind the growth of big muscles. If you're a bodybuilder who wants to gain weight as quickly as possible after training, then you'll need to make the switch to a steroid regimen, bulking workout plan no equipment. However, if you're looking for a faster leaner, stronger body in which to gain muscle, then you probably shouldn't worry too much about the differences between oral steroids and synthetic testosterone, bulking meaning in nepali. Here are some tips and questions to help you make the switch, mb mass gainer xxl 1kg. Ask Yourself: 1. Does a Testosterone Boost Help Me Lose More Weight? Yes. The growth factors in steroids can help you achieve an even greater weight loss, compared with the effects of an anabolic steroid, bulking of sand theory. A study published in the American Journal of Clinical Nutrition showed that men who participated in a cycle of high-strength training, with either oral testosterone or placebo, significantly lost significantly more pounds in the first 6 weeks. 2, bulking of sand theory. So What Exactly Does Testosterone Supplements Suppress, bulking 4 weeks? Testosterone supplements suppress growth factors, bulking hardgainer grocery. This means that if a man is on testosterone he'll notice that his muscles aren't getting bigger like before or his bodyfat percentage won't go as high as before. However, if your goal is to get the most out of your muscle gains then testosterone supplements should help you boost that growth factor production, bulking workout plan no equipment0. Testosterone and Growth Factors A hormone called GH (growth hormone) is a growth factor that is present in the body for a short period of time. GH stimulates the development of muscle mass, increases strength and fat-burning ability, as well as bone density, bulking workout plan no equipment1. It is important to note that GH and the growth factors it stimulates work synergistically to increase muscle mass. There are two types of GH we're looking to see: HGH agonists (GHA) are hormones that enhance human growth hormone release without the side effects of testosterone. HGH antagonist (GHB) are hormones that suppress human growth hormone release while still increasing bodyfat. 2, bulking workout plan no equipment3. How Should I Evaluate a Testosterone Supplements?
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The present invention relates to a pet dog shoe for location tracking. The pet dog shoe for location tracking comprises: (100) a dog shoe body; (200) a dog shoe sole; (300) a micro GPS; (400) a smartphone; and (500) a dog shoe ankle. According to the present invention, a micro GPS in a pet dog shoe for location tracking and a smartphone are interconnected, and thus the location of a pet dog can be easily recognized. Accordingly, the missing of a pet dog can be prevented.
COPYRIGHT KIPO 2021 | |
Science Grade 9
Developed as a curriculum guide for Grade 9 Science in British Columbia, Canada.
The Issue: The process of shifting what is happening in our schools toward 21st Century learning strategies and processes such as authentic and engaging learning, brain-based learning, assessment for learning, project-based learning, and systematic learning support strategies is time-intensive, expensive and slow even in schools that have made progress in becoming professional learning communities, devoted to improving learning and the assessment of learning. Wikipedia-style, broad-based collaborative development, which opens the development process to everyone in the province who has time, interest and expertise, has a tremendous potential to accelerate and enrich this effort. It may only require some group to take the initiative to get the ball rolling.
Goal: ‘Wikipedia’-style, online development to create course curricula with differentiated, SmartLearning-based lessons and learning strategies, differentiated Assessment for Learning strategies and processes, and a pyramid of intervention strategies to support both students who are not learning, and students who are learning well and need greater challenge. In time, it would be expected that there would be multiple versions of the curricula created, each meeting the course PLOs with different sets of themes, strategies and resource utilization such that the curricula would be tailored to the circumstances of a school, region, class or student.
Characteristics of 21st Century Education (link)
Recent research in Education tells us that the best schools are those in which teaching and teaching assistant staff work productively together in teams on curriculum and teaching strategies on a regular, systematic basis. We meet monthly to plan and to help to promote promising innovative learning and teaching practices at Oak Bay. The Learning Team will focus on three strands this year:
• Assessment for Learning (discussed below)
• Differentiated Learning (strategies to meet individual learning needs)
• Active, individualized Learning (strategies to make learning more flexible, individualized and engaging)
PHILOSOPHY • each individual has a natural set of potentials and talents and interests
• we learn constructively – by connecting the new with the known, in context (so ‘big picture’ questions and themes are important)
• purposefully engaging emotions positively (expression, movement, stakes, novelty, staring, apprenticeships, thinking big)
• teacher is coach – providing interesting learning opportunities, helpful feedback, and striving for max effective student self-direction (age and maturity appropriate)
• focus on individualized learning in a continuous progress model with universal access supported by tech
• we often forge meaning through social interaction and teamwork (talking, sharing and discussing)
• stories and games are important: (learning and accomplishing goals within sets of rules)
• this kind of education also naturally promotes the development of social responsibility
So, for this course, we are implementing the following:
Assessment for Learning
This involves a shift of the purpose of assessment from being primarily the way ‘to get grades’ for students, to being primarily a way of providing them feedback to help them to learn. This way, the focus is on the learning vs. the method of assessment, so that the ultimate goal is to master the course material vs. to ‘get a good grade’ (although both can be desirable).
Provincial Ministry of Education Prescribed Learning Outcomes
Active, individualized Learning with the use of performance assessments (projects): A three-year study of teaching and learning in more than 400 third-, sixth-, and eighth-grade classrooms in Chicago found that when students were given writing and mathematics assignments calling for more authentic work, they performed better on tests used to judge basic skills.
We find it helpful in developing a project to classify problems one of three ways: design a product, service, or system, improve one, or plan and stage an event. For maximum motivating power, the problem should be one adults face in a constructive field of human endeavor. When students tackle an actual problem in the community, by far the strongest learning occurs.
Link to more info on project-based learning
Effective use of Educational Technology We will also do our best to take advantage of available technology to support learning in our course: | https://en.wikiversity.org/wiki/Science_Grade_9 |
For the past several months, I’ve been collating a list of complex words I’ve used when talking to pet families. These are words that provoke a quizzical look or a clarifying question from a family member during a conversation about pet health care. In this blog post, I do my best to define this confounding terms and hopefully help you demystify some common “vet-speak”!
- Anorexia has a bad connotation from its association with the human disorder, anorexia nervosa. When your veterinarian makes a note in a medical record that says “anorexia for 4 days,” the notation simply indicates that the pet is not eating. It does not mean that your pet has been diagnosed with an eating disorder.
- Dehiscence is the term used to describe a surgical incision that has fallen apart. While this sounds serious, most incisions are multi-layered, and it’s often only the outer layer of skin that comes undone. The most common cause of dehiscence is the pet owner’s reluctance to use an Elizabethan collar, or cone, to protect the incision.
- Fasting is common enough that most pet families will understand it in principle, but it’s important to know the specifics. Does fasting mean no food AND no water, or just no food? Be sure to ask this question if your veterinarian is not clear.
- General anesthesia indicates that the entire patient is anesthetized. If anesthesia is not general it’s local, meaning just one specific site. Veterinarians don’t use local anesthesia as much as physicians do. A common example of local anesthesia is the lidocaine injection you get at the dentist.
- Incontinence describes one of two conditions: inability to control urination or inability to control defecation. Veterinarians diagnose urinary incontinence more often than fecal incontinence. The most common cases of urinary incontinence involve older female dogs who leak urine when they sleep.
- Inflammation describes the body’s response to disease or injury. The cardinal signs of inflammation were defined by the 1st-century Roman encyclopaedist, Celsus, as calor, dolor, rubor and tumor (heat, pain, redness and swelling). This four-word summary is applicable to all inflammatory processes: a healing surgical incision, the area around an infected tooth or the intestine of a patient with inflammatory bowel disease.
- Lethargy, as defined by Dictionary.com, is: an abnormal lack of energy, especially as the result of a disease. This fits the veterinarian’s use of the word to a T. When your veterinarian asks questions about ball playing or Frisbee chasing, she is trying to determine if your dog is lethargic as part of the disease process being evaluated.
- Prophylaxis is an action taken to prevent a disease. For example, heartworm medication is a form of prophylaxis given to prevent heartworms and other parasites from infecting your pet. At AMC, a “prophy” is doctor speak for a dental cleaning to prevent tooth and gum disease.
- Responsive describes a pet’s level of consciousness and reaction to their environment. The acronym BAR is short for Bright, Alert, Responsive. A waggy, friendly dog is BAR. The lowest level of consciousness is comatose.
- Undetectable, one would assume, may indicate good news. If something is undetectable, it’s not there, right? Not necessarily. Your veterinarian might use this word to indicate something they’re worried about, but just can’t find yet. In incurable diseases like hemangiosarcoma, a veterinarian might say metastasis, or tumor spread, is undetectable when they can’t find any tumor using x-rays or ultrasounds. Because all dogs with hemangiosarcoma succumb to this dreadful disease, the metastases may simply be undetectable at the time of the test.
Of course, this is an incomplete list – your veterinarian may use many words you don’t understand during your visit. If this happens, remember there’s nothing wrong with asking your veterinarian for clarification, to explain something again or to provide an example. | https://www.amcny.org/blog/2019/04/03/ten-confusing-medical-terms-your-veterinarian-might-use |
Please note: We will be closing at 2 pm on Saturday, December 24th, and we will be closed Sunday, December 25th and Monday, December 26th in observance of Christmas!
Anesthesia / Surgical Consent Form
Complete the Anesthesia / Surgical Consent Form
BOOK APPOINTMENT
Complete Your Anesthesia / Surgical Consent Form
Client Name
*
First
Last
Patient Name
*
Phone number at which owner can be reached today or tomorrow:
*
Additional number
Would you like to be contacted by text or phone call before the procedure?
*
Text
Call
Either
If text, what number would you like to be texted at?
After procedure:
Would you like us to text you after the procedure, or would you prefer a call?
*
Text
Call
Anesthetic and surgical procedure(s) to be performed:
*
I, the undersigned owner or agent of the pet identified above, authorize the staff of Russell Lake Animal Hospital to perform the above procedure(s).
*
I have read and agree
Microchip: *There is an additional fee for this procedure*
*
Yes
No
Already has one
Nail Trim: Would you like a complimentary nail trim for your pet?
*
Yes
No
Will your pet be given medications on the morning of their procedure?
*
Yes
No
What medication?
*
Other medication(s), please specify and note last time given.
When was the last time fed?
Any other concerns/allergies/procedures?
I understand that some risks always exist with anesthesia and/ or surgery and that I am encouraged to discuss any concerns I have about those risks with the attending veterinarian before the procedure(s) is/are initiated.
*
I have read and agree
I am over 18 and understand that the attending veterinarian will make every effort to contact me regarding treatment in the case of unforeseen emergencies. If unable to contact me, the staff may or may not have my permission to proceed with life sustaining procedures.
*
I give my permission [yes]
I do not give my permission [no]
I have received, read and understand the estimate provided.
*
I have read and agree to cost of procedure
While I accept that all procedures will be performed to the best of the abilities of the staff at this hospital, I understand that no guarantee or warranty has been made regarding the results that may be achieved. I also assume full responsibility for any additional expenses incurred after the surgical procedure is performed, such as follow up radiographs, re-check physical exams and additional surgery due to post-op complications. These are more likely to occur when there is a failure to comply with the aftercare instructions. I have been provided an estimated cost for the procedure(s) listed above. I assume financial responsibility for the recommended services and will provide payment in full at the time my pet is discharged from the hospital. I have read and fully understand the terms and conditions set forth above.
*
I have read and agree
I certify that I am 18 years of age or older and responsible for the financial and medical decisions for the above mentioned pet.
*
I agree
I disagree
Please type your initials. | https://russelllakevet.com/anesthesia-surgical-consent-form/ |
Don’t bother contacting me, I will be leaving all Xavier devices here when I leave.
I can’t be your villain anymore.
A former resident of the mansion and X-Man, Laurie Collins joined with Nathaniel Essex in the belief she was helping save the world and now counts the X-Men and their allies as her worst enemy.
Contents
- 1 Details
- 2 Biography
- 2.1 Phase 1
- 2.2 Phase 2
- 3 Physical Characteristics
- 4 Powers
- 5 Equipment
- 6 Trivia
- 7 External Links
- 8 Plots
- 9 Meta
Details
Character Journal: xp_wallflower
Real Name: Lauren Olivia Collins
Aliases: "Discharge" (trainee name), "Olive" (from John Allerdyce), Mutate 42, Wallflower (X-Men codename)
First Appearance: April 25, 2006
Family: Gail Collins (mother), Zachariah "Zach" Garrison (father)
Biography
Phase 1
Childhood
Laurie had an interesting childhood, mostly looked after by her grandparents during the day, and she and her mother would spend a lot of time together during the evenings. When Laurie was old enough to travel, Laurie's mother would take her on road trips to various locations around the US, and sometimes overseas to places Gail had a connection with from her modeling days. Laurie's grandparents died while she was still young, having had Gail late in life and passing peacefully in their sleep. Laurie and Gail still visit their gravestones in the local cemetery once a year.
Laurie manifested her mutant powers in her 15th year, during a summer camp where she suddenly became aware of people's changing attitude while in her presence. Upon confiding this weird occurrence to her mother, Gail cautioned Laurie to keep her new found powers under wraps considering the mood of the town.
Laurie continued with her life and schooling, only telling a few close friends of her powers until one of those friends betrayed her to the authorities after a riot at her high school. Laurie was put on trial for inciting the riot and defended by Nathan Dayspring, who eventually cleared her name. After she was released from jail, she decided to enroll at Xavier's School for Gifted Youngsters with the hope of controlling her powers and one day being able to lead a fairly normal life. The incident, however, left her with a deep distrust of the local police, and a certain hesitancy about police in general, which would later cause her issues with teammate, Garrison Kane.
Life as a 'Gifted Youngster'
Laurie quickly settled into life in the X-mansion, gaining new suite mates in the form of Jennie Stavros, Crystal Amaquelin, Angelica Jones and Sooraya Qadir, and a roommate in the form of Yvette Petrovic. She was very much a mother hen type, chosen to be Yvette's roommate due to both the calming effect of her powers and her ability to take care of the bewildered younger girl.
After a first meeting with Forge, and some discussion on her powers issues, Forge made an 'air scrubber' for Laurie which allowed her to concentrate on controlling her powers without having to worry about inadvertently using them on anyone while she learned. This marked the beginning of a friendship between the two, one that changed for a time after a Christmas kiss under the mistletoe. Despite her attraction to Forge, Laurie eventually decided that their age difference, plus the uncertainty relating to her powers was just too much of a gap to cross and ended things before they really began. This created a rift between the two which was eventually solved during the events of This Savage Land.
Ambitious and hard-working, Laurie pushed herself academically, physically and with her powers training. She wasn't all work, however, indulging in rather odd bouts of pranking to relieve the stress. She discovered, however, that her powers caused a certain degree of discomfort in people, particularly the "control freaks". When she decided she wanted to try to go without the air scrubber in the suite, Jennie reacted negatively in light of her kidnapping and mindwiping. Over time this, coupled with various incidents regarding her powers (the kiss with Forge, an interaction between her powers and Manuel's at prom which ended badly, a date with Kyle Gibney where her powers effected his enhanced senses, stopping Haller's heart by accident during a rafting trip), led to Laurie becoming cautious, almost fearful, about the impact her powers could have on others. Fortunately, this was balanced by her work with Red X, where there were several instances (Living Pele, Red X Mission: Whiteout, Red X Mission: Rising Waters) where her powers were beneficial.
When she found herself kidnapped along with the many of the students, again she was able to use her powers to help herself and her friends. The experience shook her up, however, and she returned to her mother's house until Jennie came to talk her into coming back. The apparent deaths of Crystal and Forge with the disappearance of Attilan hit Laurie hard - unable to do anything to help, she became uncharacteristically angry and moody, having several journal altercations, especially with the equally hot-tempered Kyle.
Floundering
An incident in a dream world led Laurie to start wondering about her absent father, a man she'd never met and who her mother rarely spoke of and never in a flattering way. When Laurie's mother fell ill during a morning taping, Laurie discovered that her mother was suffering from cancer brought on by a prolonged exposure to Zach Garrison's mutant power. After Laurie hunted him down, Zach eventually (and reluctantly) consented to help them find a cure for Gail, and he moved to New York so that he and Laurie could reconcile their familial relationship into something resembling that of a father and daughter bond. Gail was less than pleased with the situation, but couldn't deny Zach access, despite his less-than-rosy past.
Following Gail's cure, Laurie became an X-Man trainee, more determined than ever to fix the world. Her enthusiasm led to a rift opening between her and her roommate, as Yvette felt more and more treated like a child and a victim than like a friend and peer. All was not well on the team either, with Laurie's insecurity regarding her powers and her inability to trust others causing issues with her teammates. She acquitted herself well in Japan, engaging Noriko's help in resuscitating Julio Richter after he stopped a major earthquake in Tokyo. It was a small break, however, as the chaos caused by the breaking of the astral plane drove Laurie to push herself into taking care of everyone around her, whether they wanted it or not. A threat to use her powers to force Jack to eat was taken extremely badly, but Laurie didn't seem to realise what she was doing until a talk with Jean Grey, who she respected immensely and who had also trained with her father and seen the results of using pheramone powers recklessly, frightened her into physical flight. Trying to repress her fears, she engaged in a series of pranks that tended to annoy rather than amuse. Only Morgan Lennox, her new suitemate, seemed to appreciate her.
A series of 'failures' followed. Her training with the X-Men faltered, as she became obsessed with being a 'good' X-Man and compared herself to everyone around her. Her hand-to-hand classes with Garrison were curtailed when she used her powers on him without telling him. She failed to get into NYU's medical school and attempts at "helping" were more often interfering, such as her telling Jay Guthrie about Kevin's powers addiction. Then a mission in Sri Lanka went terribly wrong as the Indian military conducted ethnic cleansing of Tamils trapped outside the Red Cross camp and the Xavier's people were forced to watch. Laurie, with her need to "do something to help", found the situation especially intolerable, at one point restrained by Crystal from taking on the Imperial Guard. The incident shook her up greatly and she returned to the school in even more turmoil than before.
Hitting the Bottom
The next two months were marked by the loss of two friendships, seemingly for good. Injured when trying to stop Yvette from attacking the man who had kidnapped her, Laurie felt betrayed that a friend would hurt her, even in the throes of emotion. And then in October 2008, following an attack by another mutant, Laurie accused Kyle of not being able to control himself due to his mutation on the open journals, causing an eruption of open hostility against her. Alienated by almost everyone she had once called 'friend', Laurie fled home once more, only to have her mother send her back to the school to take responsibility for her actions.
Apocalypse's attack on New York pushed Laurie to limits that terrified her. Fleeing with her father, she abandoned him when he 'stole' from the apartment they had sheltered in, only for him to come to her rescue against Post. When Post nearly killed Zach, Laurie saved his life, and later discovered she had the reserves to control entire crowds of people as she coordinated with Team Alpha against War. This discovery, plus the knowledge her stubborn moralism had nearly cost her father his life, pushed Laurie even further into not-coping. Eventually she wound up with pneumonia from self-neglect, and wound up in the sick bay and then bed rest for a time.
Laurie Collins World Tour '09
Pushed beyond her limits, physical, mental and emotional, Laurie decided a change was in order, and hit on a plan to spend several months traveling the world. Starting in Ireland with Morgan, she met the former mercenary's team mate Eamon again, the two of them hitting it off. Then, joined by Crystal, she continued through Europe, meeting up with mansion residents along the way in Germany, Egypt and Bosnia-Herzegovina.
The trip did Laurie much good, although there were the occasional lapses in her judgement - one or two further journal altercations with Forge and Kyle, a heated email exchange with Manuel about his wanting to keep Laurie (and her powers) away from his empathic sister Valentia. Meeting up with Eamon again in Venice, she incurred Morgan's wrath by losing her virginity to him. It wasn't a one-time thing, however, with both of them deciding to give a relationship a shot despite all the obstacles surrounding it, not the least of which being Eamon's continuing work as a mercenary.
In Egypt she found herself, along with Crystal and Kurt, facing down a possessed Monet St. Croix - Laurie used her powers to subdue the archeologist using Monet as a weapon. In Bosnia, at a Red X venture, she reunited with Dr. Pierre Marcel, the man who had stitched her up following the incident with Yvette in Kosovo. Marcel is a long-time member of Medicans sans Frontiers, and was able to give Laurie much advice and support, especially in regards to the genocide in Sri Lanka. The peace was shattered, however, when Yvette was injured by a land mine and evacuated to hospital in Kosovo. With doctors unable to treat her due to her mutation, Laurie attempted to use her powers as she had before, to make the younger girl relax, but found she was too far gone for Laurie's powers to work. It was difficult, accepting she wasn't able to do anything for her once-roommate, but Laurie managed far better than she had in the past, looking after others without being obnoxious or holier-than-thou about it.
Following the incident, Crystal and Laurie traveled to Dubai, where they were to meet Forge, Kyle and Doug for a vacation in a state of the art electronically-controlled hotel. After some initial confusion about Laurie's presence (and some rudeness on Forge and Kyle's parts), the group settled in nicely, until attacked by Milan and Quentin Quire, nerds with grudges against Forge and Doug. Laurie and Kyle were trapped in an elevator but managed to escape, Laurie using her powers to give Kyle the energy to climb multiple stories with her clinging to his back. She then joined Crystal in taking down Quire, although he managed to attack her psychicially for a moment before Crystal bounced him off the wall. Regrouping and relaxing in Attilan briefly, the group then returned to the school, where Laurie announced the end of the trip and her hopes to complete her X-Men training and get back on track with her attempt to get into pre-med. New suitemate Jean-Phillipe Colbert was in for a shock.
A New Leaf
Laurie began July 2009 determined to start afresh, clearing the air from old disagreements and burying the hatchet with those she'd fought with. The new beginning was almost literal, when an incident at an art exhibit resulted in a powers swap and Laurie manifesting Crystal's ability to control air molecules, while Kurt Sefton acquired her pheromones. Laurie was perhaps less upset about the new powers than the others, setting herself to train with them and mourning their loss when her own 'morally dubious' abilities came back, something which disturbed Crystal.
Continuing her medical studies and working in the medlab, Laurie began to use her powers more to assist healing, a reluctant Nathan Dayspring her first patient. She also returned to the X-Men, the training a lot harder than before after her prolonged break. She still had issues, however, conflicted about how she should act with various people when faced with differing opinions, and her relationship with Manuel was as still as volatile as ever, given his protective streak over his sister, to the point they sought mediation from David Haller.
Two relationships she did salvage were with Yvette, her former roommate and fellow trainee, and with Logan, although they would never be friends. With Yvette's mediation, Logan and Laurie got to the point where she accepted stealth training from him, joining Yvette in the work she did with the older X-Man. She also got pointers from X-Forcers Amanda and Doug, about teamwork, trust and combat, before they took her to Silver in an effort to get her to relax from her usually tightly-wound self.
Following the mission in Pakistan, during which she and Hank worked together to save refugee lives and during which she prevented one of the students, Fred Dukes - Phase 1, from killing a member of the Imperial Guard, Laurie finally obtained her blacks and took the X-Men codename of 'Wallflower'. It was a very proud moment for Laurie, who had felt (and sometimes been told), she would never graduate past trainee. Her first 'mission' was the graduation attack, where she was aided by a mysterious mutant (who turned out to be Crystal in disguise).
X-Factor
Laurie's close friend Vanessa (whom Laurie still called Morgan, as that had been the name she'd previously known her as), and Bishop set up a private investigation service in District X, with funding from the Xavier Institute. Laurie volunteered her services as a personal assistant, although she was quickly 'traded' to Bishop as Vanessa could never find anything after Laurie had 'tidied' her desk. As an unofficial part of the group, Laurie was involved mostly in the paperwork side of investigations, such as the missing persons case that was the group's first. She took the returned Jean-Paul Beaubier under her wing also, confused by the changes in his personality and behaviour since his departure from the mansion the previous year.
She continued to work with both Red X and with the X-Men, assisting in the evacuation of wildlife during a wildfire in Alberta and in X-Men missions such as the search for a pyrokinetic firebug in Chicago. Laurie experienced first hand the effect of an inhibitor on her body's chemistry, suffering from cramps and disorientation while under the effects.
Perhaps due to the influences of Vanessa and her boyfriend, Eamon, Laurie herself began to relax and act more naturally, even getting drunk at a college party and calling Kyle for a ride home. The pair also revisited their worst moment when Karl Lykos resurfaced and became a investigation for X-Factor; recognising the modus operandi from the coroner's reports, Laurie and Kyle lent their experience and training in dealing with Sauron to the team, helping take him down for SHIELD to take into custody.
With Eamon working in New York for the duration, Laurie's life seemed to be on the upswing, her roommate Jean-Phillipe commenting that she was far too cheerful from all the sex she was getting. Her sex life became rather more public than she could have anticipated one night in March 2011, when her air scrubber failed while Eamon was staying over at the mansion; the ensuing spread of pheromones impacted the graduate wing of the building and resulted in several assignations and a particularly bizarre situation where Marius Laverne jumped out of a window to avoid having sex with Laura Kinney. Shocked and embarrassed, Laurie fled to Eamon's apartment until the filter could be fixed, reacting defensively on the journals. At last, she came to her senses and apologised for the inconvenience, especially after several emails from her friends.
Laurie's lack of people skills reared its head once again in the X-Factor offices, when the grieving friend of a murder victim came to demand answers. It perhaps wasn't Laurie's best performance and that summer, she decided another break - shorter than the World Tour - was in order. With a group of friends - Kyle, Kevin, Laura, Angelica and Jean-Phillipe - she left the mansion for several weeks for a road trip to New Orleans, returning in early August 2011.
When Yvette's eighteenth birthday celebration was gate-crashed by Toad and Mister M., who released Kick into the air, the drug set Laurie's powers off, and she cast a bunch of happy vibes that mixed with Megan's pixie dust, creating a rather happy atmosphere within the club. The resulting headache the next morning, after the high had worn off, didn't take away from the fun, at least.
Life stayed busy for Laurie, between X-Men training and school and just in general, resulting in her dropping a class at school and quitting her job as Girl Friday with X-Factor.
2012
The year started on an interesting note when a film festival, featuring an independent movie about the events of Day Zero, was targeted by terrorists. Laurie joined the security detail, which included a fight between mutants and FOH protestors that she helped to diffuse.
Much like the rest of the world, Laurie kept a close eye on the news when word of Genosha's crime against mutants started becoming public knowledge. She, along with a couple other younger X-Men, acted as bait in a sting to capture mutant traffickers in an attempt to find out more about what was happening in the country. In May she received back her MCAT scores, and was disappointed by the results, though they were still quite high, and enough to get into NYU for medical school.
While attending a protest at the Genoshan embassy, Laurie was kidnapped along with many other members and associates of Xavier's. She ended up in Genosha, stripped and powerless, and spent a night in a cell suffering from the effects of losing her powers before being put through the mutate process along with several other Xavierites.
Following the botched rescue attempt, Laurie - as Mutate 42 - and her handler, Semoko, ambushed a group of escapees at the Genoshan Broadcasting Commission building, and later ambushed a second group attempting to take out the tank laager. It was there that Mutate 42 was finally subdued, and the Xavierites were able to reverse the mutate process, bringing back everyone who had been put through it. She assisted in the final attack and take down of the monstrous Thomas Moreau.
Returning home and back to normal was a difficult path for Laurie. She spent a lot of time feeling scared, and was very obviously (to anyone who talked to her) not okay. She blew up at Matt after he disappeared and subsequently posted on the journals laughing about his punishment, though she later apologized. Not long after her return, Eamon came for a visit, which did much to help Laurie's spirits. She went away on vacation with him in the middle of July, returning a few weeks later.
In August she volunteered herself as security at a mutant-positive rock concert being threatened by the Purifiers, lending her abilities to keeping the crowds calm when an attack occurred. She started medical school in September, unhappy with having to start at a new school but making it through without much difficulty.
2013
Laurie rang in the New Year with a hangover, spending a good deal of the first few months buried in books and homework, resurfacing every now and then to let people know she was alive. In February, she joined a group of X-Men in rescuing a shipwrecked Adrienne and Garrison from Magneto.
The year passed mostly quietly, with Laurie throwing herself into her studies, often losing track of what day it was. In August 2013, she and Eamon broke up, and in September, she and Doug discussed the possibility of going out on a date, although it didn't actually happen until after a horde of demons invaded the mansion in December.
2014
The year started on a quiet enough note, with Laurie and Doug continuing their relationship at their own pace, managing a couple more dates between school and general life responsibilities. August brought with it an overload of work in the med lab when a team of X-Men were badly injured on a mission, and in September she helped Kyle rescue a young mutant who was playing vigilante on the streets of Brooklyn. Things took a bad turn for Laurie at the end of the month when she was kidnapped along with Hope Abbott, and held hostage for several weeks where she was tortured, mentally and physically, for information about the X-Men. Upon being rescued she was nearly catatonic and in a state of acute sleep deprivation.
It's The End Of The World
Laurie's recovery went well, and by the January 2015 she was ready to join the X-Men in defending Muir Island when it came under attack by the Brotherhood. The fight took a turn for the worse, and Laurie's arm was cut off Psynapse. She spent the next few days drugged in the med lab, and when the mansion came under attack by The Dark Phoenix North had to force her to keep moving and evacuate.
Phase 2
A Whole New World
Despite all odds, Laurie was among those to wake up in the new universe created by Xorn after their old universe fell apart. Unfortunately, she was still an arm short. She made the best of it, however, throwing herself into her work in the med lab and spending as much time as possible with her friends and Doug, eventually moving in with him. She did quit the X-Men, acknowledging that one arm wouldn't make her useful in a fight.
She began looking into prosthetics and trying to make life as easy as possible on herself, throwing herself into mansion life. She also began looking into returning to medical school, though she wasn't sure yet how she'd be able to make rotations work with a prosthetic arm. Still, it was a quiet year for Laurie, exactly what she had earned after the way the year had started.
2016 was similarly quiet - Laurie went back to work and school, resuming her normal life. She also threw herself once more into taking care of her friends and partners.
Sleep deprivation and work ruled most of 2017, although Laurie did find herself recruited to the new team Clint Barton was starting, acting as the medical expert for the team.
Time Passes
Laurie kept herself busy with work, working through different rotations at a nearby hospital. She also continued to hone her less than welcoming personality, largely withdrawing from mansion life. That didn't exempt her from getting involved in missions and mansion shenanigans, of course. 2018 was particularly eventful - she joined a group going to rescue Christopher Summers and his crew from mind-controlling jellies (and thankfully did not have to join in on turning into a sea creature to help), assisted with first aid when a fire android attacked NYC, and was drawn into an odd, zombie-esque scenario during a Danger Room mishap. 2019 was mildly quieter - magical hammers were set loose around the world, and a cursed house pulled Laurie into her worst nightmares, but it could have been worse. 2020 passed in blissful peace.
Time Changes Everything
Laurie's year started much like the last had, quietly and with work as her focus. However, when nanites attacked the mansion as part of a scheme by The Fixer, Laurie lost the use of her advanced prosthetic and began a year-long journey that would see her manipulate her nearest and dearest and burn all her bridges. This downward spiral would culminate in the events of Arise, X-man and force Laurie into the company of Dr. Nathaniel Essex. Where she'll go from here? Only time will tell.
Physical Characteristics
Height: 5'8"
Weight: 115 lbs
Eyes: hazel
Hair: brownish-blonde
Other: When exerting herself with her powers, Laurie's hair, skin and nails all turn purple. This can take several hours or days to fade, depending on the depth of the power use.
Powers
Laurie's mutant power is that of pheromone control. She can affect the physical state of a person through the release of these pheromones. They do not effect a person's emotional state, although if someone did not know what was going on, they might develop an emotional reaction such as fear simply from the strangeness.
An example: when Laurie is in a state of stress, people around her will start to sweat, their heart rate will increase and they will have tightness in their abdomen but they will not have the corresponding emotion for this set of reactions. It would be very disorientating to someone who didn't know what was going on.
One of the side effects of her power makes her a clear communicator with animals, although she is not able to understand them at all. This has also been shown to be detrimental, as in the dinosaur incident, where she discovered reptiles and insects are drawn to her.
As of 2009, Laurie is now able to control whether her powers are "on" or "off", including when in contact with someone, although she needs to wash her skin to avoid residual contamination. Under great stress she is still known to "leak", and if she has been using her powers a lot, she turns purple. She has also been shown to be working on limiting the 'on' state of her powers to particular areas of her body.
Equipment
Formerly the air scrubber Forge made for her. Looks vaguely like an iPod and makes a small humming noise when turned on. She uses it mainly now when sleeping. Laurie also owns a bow that Forge enhanced for her after she used one made from his leg during the dinosaur incident. While she doesn't use it in combat, she has become a crack shot with the weapon after several years of practice.
Trivia
Laurie used to be on the track team at her old school and still keeps up her training in the mornings, usually getting up quite early to do so.
The name of Laurie's journal is inspired by the song Where the wild roses grow sung by Nick Cave and Kylie Minogue
External Links
Phase 1 Comms
Plots
Phase 1
2006
Living Pele (as part of Red X)
2007
2008
Red X Mission: Shake Hands With The Devil
2009
2010
X-Men Mission: Who You Gonna Call?
2011
Mortimer Toynbee's Electric Funkedelic Boogaloo
What Goes Around, Comes Around
X-Men Mission: X-Mas in Milwaukee
2012
X-Men Mission: Day Zero, the Movie
X-Men Mission: Amid These Storms
2013
2014
2015
- A Touch of Brimstone
- The Dark Phoenix
Phase 2
2017
2018
2019
2021
Meta
Socked by: Seraph for the Mods
PB: Emily VanCamp
Laurie was originally a played character, brought in as a student in 2006 and played by Seraph until 2021, when she left and became an NPC as a result of the events of Arise, X-Man. | http://www.x-journal.net/Wiki/index.php?title=Laurie_Collins |
This invention relates to the electrostimulation of muscles, for example paralysed muscles of an at least partially paralysed patient, to promote movement for example walking, standing or sitting.
It has been well known for many years that muscles stimulated with electricity can be caused to contract and this subject is well documented. My U.K. patent specification No. 2 136 297 describes equipment for stimulating the legs and hips of a paralysed person to produce a smooth contraction of the muscles to enable that person to walk.
However, patients who are paralysed above a certain level require the use of postural muscles to enable them to balance and to maintain an erect posture. Since the body itself is a conductor of electricity it has proved extremely difficult to avoid the stimulation of one muscle group interfering with the stimulation of another muscle group whether the muscles are adjacent each other or remote. In fact,the further apart the muscles are, the more complex the interference is. Also, if the muscles require substantially different levels of stimulation, then a potential difference is set up through the body which may stimulate any intervening muscles between the two stimulated muscle groups. For example, it is required to stimulate the quadriceps for extension of the knee and the gluteals for extending the hip. Interposing is the illiopsoas muscle which flexes the hip. Since the illiopsoas (or psoas) is an extremely strong muscle it picks up the stimulation between the quadriceps muscle and the gluteals and flexes the hip as well as reducing the stimulation to the quadriceps, causing the knee to give way and the patient to fall to the ground.
I have now devised techniques for electrostimulation of the desired muscles, which overcome the above problems.
In accordance with this invention, there is provided an electrode harness arranged to be worn so as to place electrodes of the harness in contact with appropriate parts of the body for stimulating the quadriceps and gluteals, and electronic apparatus for providing electric currents to the electrodes to stimulate the quadriceps and gluteals, said electronic apparatus including circuits which are electrically isolated from each other and include the electrodes respectively for the quadriceps and glutealts.
With this arrangement, the quadriceps and gluteals are stimulated as required without any unwanted stimulation of the psoas also occuring.
Also in accordance with this invention, there is provided a method of electrostimulating the quadriceps and gluteals of a patient, comprising supplying electrical currents to electrodes appropriately placed in contact with the body, these currents being supplied over respective circuits which are electrically isolated from each other so that unwanted stimulation of the psoas is avoided.
Figure 1 is a diagrammatic representation of the lower part of the human body;
Figure 2 is a circuit diagram of the equipment;
Figure 3 is a diagram to illustrate the distribution of electric pulses to the muscles;
Figure 4 shows a harness for one leg; and
Figure 5 shows an electrode belt.
An embodiment of this invention will now be described by way of example only and with reference to the accompanying drawings, in which:
Referring to Figure 1, there is shown the spinal column 1 and pelvis 2 of a patient. One leg and its muscle system is shown, namely the psoas muscle 3 attached to the spinal column and the top of the femur and is used for flexing the hip, the gluteal muscle group 4 which is attached to the ileac crest at the top of the pelvis and the back of the femur 5 and serves to extend the hips, the quadriceps muscle group 6 which is attached from the top of the femur through the patella 8 to the tibia and is used for extending the knee, the hamstring group 7 attached at the back of the femur and the back of the tibia and used for flexing the knee, the gastronemius attached to the femur and to the heel, and the soleus 11 attached to the tibia and the heel: the gastronemius and the soleus are both part of the plantaflexor group and are used for extending the ankle. The dosiflexors 12 are also shown and are attached to the tibia and foot and serve for flexing the ankle.
In order to enable a patient to stand it is necessary especially in the higher lesions to stimulate the gluteals, the quadriceps and the plantaflexors. In accordance with this invention, the gluteals and quadriceps are stimulated over circuits which are isolated from each other, to avoid a short circuiting of the stimulation pulses through the psoas (which,being strong, would react immediately and cause the hips to flex instead of extend), and a reduction of power to the quadriceps and collapse of the knee.
Figure 2 showsin particular the pulse generator employed, This includes a battery 31 (usually 12 volts) and a battery charger 32 powered from the mains. The battery supplies an integrated circuit 35 (which generates the basic stimulation pulses) via an ON/OFF switch 33 and a fuse 34. The pulses from circuit 35 are adjustable for pulse width and pulse rate by potentiometers 36,37 between the ranges 0.2 and 1.8 milliseconds and 10 and 40 Hertz respectively. The pulses from circuit 35 drive an output stage integrated circuit 38, which in turn drives Darlington-configured transistors 39. These drive transformers 40, providing two separate and isolated outputs 41, one for the posture and one for the leg muscles.
Figure 3 shows the distribution of pulses from the two isolated outputs 41 to the various electrodes. For one leg, there are electrode pairs G1, D1, P1 and Q1 serving the gluteals, dorsiflexors, psoas and quadriceps, respectively of that leg: for the other leg, there are corresponding electrode pairs G2, D2, P2 and Q2. For each leg the gluteals and dorsiflexors electrodes receive their pulses from one of the outputs 41, and the quadriceps and psoas receive their pulses from the other output 41. The pulses are distributed to the muscles through respective emitter-follower circuits, shown diagrammatically, with individual potentiometers providing control.
5
Figure 4 shows a harness assembly for one leg. It comprises bands 30-33 carrying electrodes 50-57 and connected together by two straps 22, 23 running the length of the leg, with a strap portion 24 to pass under the instep. The bands are provided with fastening means 34 (e.g. "Velcro") so that they can be secured around the leg and place the electrodes in contact against the body in the correct positions. Wires to the electrodes are sewn into the straps 22, 23 and the electrodes are shown as follows: 50 and 51 for the quadriceps, 52 and 53 for the hamstring muscles, 54and 55 for the dorsiflexors, and 56 and 57 for the plantaflexors. This harness is as more fully described in my U.K. patent specification No. 2 136 297. Figure further shows an electrode belt to be worn with a pair of the leg harnesses. This has electrodes 61,62 for the right and left psoas and the belt is worn so that these are positioned at the back, either side of the spine. Smaller electrodes 63,64 for the right and left psoas are provided to locate on the abdomen either side and slightly lower than the navel. The gluteals are stimulated by electrodes 65,66 on the belt, placed on the back either side of the spine and just below the iliac crest, and paired with the electrodes 52 at the top of the leg harnesses. | |
Do you value inclusion as part of health and well-being at work, at home, in relationships?
Is intercultural development and growth something you aspire to enhance as an individual and/or as a leader of your team/ organization, teacher of a class, or parent of a family in 2023?
Is inclusion a high value in your life, in your relationships, whether at work, in private, or at home?
Are health and well-being the area of healthy relationships something you'd like to focus more on in your life in 2023?
Intercultural development and growth is a journey that can help to nurture the value of inclusion and healthy relationships.
But before I dive deeper into the topic, let us take a quick look into what is culture.
What is culture?
According to the Intercultural Development Inventory (IDI) developed by Dr.Mitchell R.Hammer, professor of International Peace and Conflict Resolution, culture is all about cultural patterns, values, practices, and behaviors.
In the IDI context, it is not talked about cultural specifics such as gender, race, ethnicity, religion, and nationalities - yet those specifics inform our cultural development and growth. Our cultural expansion is about something other than learning, for example, how to exchange business cards in Japan or how to use chopsticks, although those are valuable experiences.
It is more about the cultural and behavioral patterns and getting to know and understand them under the iceberg's surface.
Imagine an iceberg. Culture is the whole iceberg of every objective cultural part on the surface, including all subjective cultural aspects under the surface.
It is about developing a deep understanding of Self AND others!
How do we identify with cultural differences and commonalities?
Often I hear clients, friends, family members, and colleagues saying I strongly believe we are all human - we are all the same.
It just recently occurred again in an IDI debriefing session with a client.
It is a belief that many carry along taught either at school, home, family, university, church, or any kind of educational system.
Tough Love Talk on the sentence "we are all human"
In the next couple of sentences, be prepared I will be more clear and direct with my language and expression - like tough love talk.
Before a quick reminder, when I talk about cultural development and growth, it is not about right and wrong!
Keep that in mind!
We can't be inclusive if we do not value the differences and diversity within and outside of us in our environment and in the people surrounding us.
This belief that we are all human is not helpful regarding the value of creating an inclusive world. Period!
It goes along with people being colorblind and not seeing white as a color.
To break it down, neither is this belief helpful in our relationship, whether with a friend, colleague, sibling(s), parent(s), partner, spouse, clients …
It is not!
I'm reminding you here again cultural development and growth are NOT about doing it right or wrong!
Because what often happens when we reflect on our development, our sweet mindset tends to function in a judgmental, sabotaging way, and - our mindset easily creates negative sentences in us and feelings of doing it right and wrong.
I know what I'm talking about:-)I have been there many times.
Here is the truth it is about development and growth - cultural development and growth - deepening and expanding cultural Self and raising cultural other understanding.
Along that way of growing into an intercultural mindset and cultivating growth, we will stumble, get discomfortable and experience setbacks many times.
That's normal and simply part of the package.
I have been there, and many people have been there, and that's simply part of learning and growing, deepening and expanding.
Not about getting it right and doing it perfectly.
Nope. It does not work that way.
And guess what it is also joyful and fun if we walk the path with heart and humor and grow our cultural competence for the sake of creating inclusion.
Okay, I'll stop the tough love talk here now.
Back to inclusion and the often used sentences such as" we are all human" or "when I'm with people of all kind, I only see us as all humans" - means I only see the commonalities between human beings.
Commonalities AND Differences
It is great to notice commonalities.
And only seeing commonalities can be hurtful and painful. Be aware of that. When we are nurturing only that belief, it creates harm within us and other people.
Why?
Because we diminish and minimize the potential and value of our differences!
Our differences AND commonalities are equally essential to create inclusion and healthy relationships.
If we see and fear differences as a threat, we tend to focus only on commonalities.
Guess what happens?
If we only focus on commonalities, like in the example I mentioned above, "we're all human," we mask and oppress differences and cut out an important part of an individual human being.
It is like not seeing and valuing the whole person. Like a cake where you take a piece away and do not see it as part of the entire cake, that makes it whole.
Let me give you another example.
Let's say your partner/spouse, and you both value a bright and light home with many windows. That might sound superficial, but underlying the value of a bright home is a deeper meaning for both. Maybe it will lift you both up, and you will get less depressed. Okay, that's what you share in common. Now let's say one of you values a wooden floor but no carpet, and one prefers carpet. That difference might look at the first objective, but the underlying might be a behavioral pattern.
Means what?
The reason why the person values a wooden floor may be because the wooden floor creates a feeling of calmness and groundedness, and overall it has an impact on the shared home because that person, in a calm way, acts in his/her best version.
In contrast, the person who appreciates carpet may associate feelings of warmth, coziness, love, and being held, and may create this feeling in the shared home, which ultimately serves all in a good way.
Makes sense?
Now we have the commonalities of the bright light windows in the home and the difference of valuing different floor textures that holds different potentials of creating feelings with varying qualities in the environment/home.
Interesting isn't it?!
Can you see how valuable it is to focus on differences instead of swiping them under the surface?
We may mask differences with valuable potentials and perspectives that help us to create beautiful environments.
It is not only the commonalities that connect us but also the differences that build deep connections, inclusion, and healthy relationships.
The differences within us and between us have true potential - the potential to create an inclusive world, family system, teams, communities, and organizations.
We need to start, continue seeing the difference, and acknowledge each other our differences!
Opportunities of cultural development and growth and honoring the value of inclusion
The IDI assessment and debriefing are a great way to enter the realm of intercultural development and growth. It helps to evaluate where we are on the continuum of growing from monocultural to intercultural mindsets at home, working places, in relationships, and in the world.
It supports us in learning about how we navigate cultural patterns of behavior.
Please reach out if you want to work on intercultural development and growth as an individual, as a family or team, or as an organization. The IDI assessment, debriefing, and coaching have truly become a passion of mine.
I would love to be of service and support you on your intercultural journey of growth and walking the pathway from a mono mindset to an intercultural mindset.
Reach out to me and send me an email at [email protected] or drop a comment below.
Happy new year 2023! | https://www.evocativehealthpath.com/post/template-product-review |
From Benedict Anderson to Marshall McLuhan: Nationalism and How Memes Could Prevent World War 3
Ninety-five years ago this month, the armed hostilities of World War I came to an end with the signing of the Armistice of Compiègne between the Allies and Germany. World War I exposed to the world the horrors of industrialized war and the conclusions of hyper-nationalism, a fate which would repeat itself with a greater fury fueled by ideological warfare in 20 years’ time. Can we, today in the age of the internet, envision such a global war happening again?
Destroying itself twice in the 20thcentury, the possibility of Europe at total war with itself in the 21st century seems unlikely to most people –Infinity Ward excluded. There are many reasons that can be listed; the structure of the EU, the memory of the world wars, the interconnectivity of the world market, the trend of military size reductions among developed states, the shift in the nature of warfare, the still present possibility of mutually assured destruction via nuclear war. Another factor we’re going to consider listing in this week’s blog is the possible challenging of nationalism.
The debate on the relevance and efficacy of nationalism today is far from being settled. The recognition, however, that economic and political ties have expanded beyond state boundaries is undeniable, as is the recognition of social and cultural identities expanding beyond direct geographical and ethno-centric boundaries. As tends to happen in academia, there’s a wild orgy of terms to describe these processes: globalization, regionalization, transnationalism, internationalism, supranationalism, post-nationalism. We’re going to approach this topic by starting way back, with the internet’s ancestor –the printing press.
Social theorist and post-colonial pop sensation Benedict Anderson has put forth a great theory addressing the origin of nations in his widely-praised book Imagined Communities. Anderson argues that nations are not communities in the traditional sense because “the members of even the smallest nation will never know most of their fellow-members, meet them, or even hear of them, yet in the minds of each lives the image of their communion” (Anderson, 224). As such, the community and the affinity of its members that is perceived by the individual is an imagined community and affinity, or a socially constructed one.
Nations, moreover, are different from religious communities which can also be understood as socially constructed communities. The critical difference between these two is in the structure of the relationships between the community members. Nations, Anderson specifies, are sovereign. Formulating during a period in history where the belief that the only source of authority originated from external divinity was being challenged by ideas that held the human being to be a source of authority based on inherent nature and self-evident rights, the sovereignty of the nation is perceived as inherent in and of itself. This not only establishes the legitimacy of a nation in face of other nations, but also, importantly, establishes the nation internally as a horizontal community where, despite socio-economic inequalities, all members are united by a deep horizontal comradeship. A child may be born unequal in wealth and status in comparison to other members of her nation, but she is born equally British. This relationship based on horizontality is what strongly distinguishes the nation from preceding types of communities; the shift from a religious community to a nation marks the shift from subjects to citizens. Now that we have our definition of a nation, we can ask: how did the nation come to be imagined?
A missionary printing press.
Anderson’s answer is print capitalism. 150 years after the invention of the Gutenberg press, and around 425 years before 50 Shades of Grey, the markets for books, which were restricted to Latin, had become saturated. To keep business running, new, much larger, markets were sought –those of the vernacular. This process was supported, and to an extent preceded, by religious reformations, specifically Protestantism, which reproduced religious texts in the language of the locals. Print capitalism, however, due to the massive scale of its production, made it possible for speakers of a same language, who could not converse before because of the differences in their dialects, to communicate with each other via print. Because books could last for generations, print capitalism also slowed the rate at which languages changed, making it so that the language used decades ago was very similar to that used in the present. In effect, print capitalism in the 16th and 17th century consolidated the speakers of a language by overcoming the differences in their dialects and by creating a sense of continuity over time. This created what Anderson refers to as ‘language-fields’, or geographic areas in which inhabitants could communicate and identify with individuals who lived far away or who they would never even meet. Here we see how membership to an imagined community is formed, and how print media can determine the boundaries between ‘us’ and ‘them’. But how did print capitalism cast membership as horizontal? Let’s get meta-physical.
Anderson argues that the imagining of the nation could not happen without a fundamentally new perception of time. Whereas members of a religious community perceived time as related to divine providence, where the past, present and future were all connected to a deity and so were all merged into one, individuals of the 18th century came to perceive time as ‘homogenous, empty time’, a notion Anderson borrows from German philosopher Walter Benjamin. Homogenous, empty time is time as we perceive it today; time of empty, linear, calendrical days which we all simultaneously act in and occupy; time in which the past, present, and future are laid out flat horizontally in a line and distinguished from one another. The perception of homogenous, linear time, Anderson theorizes, was induced, although not exclusively, by two important inventions brought forth by print capitalism in the 18th century; the novel and the newspaper.
The novel presented narratives to readers in a new way which emphasized simultaneity. Novels now told stories of fictitious members of real societies simultaneously carrying out their own sequence of events -character A walked to the market, B talked to C in a dining room, and D arrived at port, all unaware of each other’s existence yet tuned to the same clocked, calendrical time conjured up in the mind of the reader. The flow of the world constructed in 18th century novels mirrored the world imagined and experienced by the individual of the 18th century. Newspapers, on the other hand, were not only a catalogue of local and global events happening simultaneously in calendrical days. They were in themselves a social practice unique to homogenous, empty time.
Every morning in silent ceremony, Anderson explains, each individual reading the latest print would sense, or imagine, herself in communion with her fellow citizens as they all read the latest news about their community, identifying with individuals she would never meet or know. The relationship experienced here between individuals was temporally horizontal; all members of a nation occupied the same temporal plane. Whereas members of a religious community where related through their relationship to the same deity, they were related as subjects to a higher holy being above, members of a nation where related through their simultaneous movement through linear empty time. Hence why the shift from religious community to nation is a shift from vertical relationships to horizontal relationships. Importantly, because time was perceived as linear, it became possible for ideas of progress and notions of forward movement through history to develop. Thus, the mission of a nation to progress and move forward became a new purpose uniting members of a nation not just with one another but with their national ancestors and descendants. In effect, print capitalism induced the forming of nations by consolidating locals of a geography with a common antiquated language and with a simultaneous experience of moving forward through homogenous, linear time.
Does the internet function in the same ways mass print did to create global imagined communities? Meme culture can seem to indicate so. The internet has created a new language of memes which not only takes procrastination to a whole new level but also overcomes differences in spoken and written languages. Memes and rage comics can be found in several languages, effectively communicating stories about real life experiences. Not only do memes and rage comics allow individuals from across the globe to communicate with each other, it presents the story communicated as a universal experience, further strengthening identification among individuals from different nations. Like novels, they communicate stories about real individuals in real societies simultaneously living out their lives. Amplifying the effects of reading a newspaper, the act of communicating through memes itself is immediately simultaneous, allowing you to see the post rise and drop in popularity, as well as view and engage in live conversations about the post. It’s no surprise that many user-generated content websites, like reddit and Tickld, have dynamic self-conscious communities. But is it so simple? Do new media technologies just carry on the developments of print media? Or are new currents underway today?
Marshall McLuhan, “The medium is the message”.
Marshal McLuhan argues the latter. McLuhan was a Canadian philosopher, media guru and intellectual celebrity who you can’t help have a love-hate relationship with. McLuhan is known for coining the phrase “the medium is the message”, popularizing the notion of “the global village”, and for pretty much revolutionizing the way we look at media’s role in shaping human societies. He’s also known for his “eccentric” writing style which leaves you wondering whether you just read the most mind-blowing idea ever or something that should be backdropped with really high guy. McLuhan, who is credited with quotes such as “diaper spelled backwards is repaid. Think about it” and “I don’t necessarily agree with everything I say”, is also credited with predicting the internet thirty years before its commercialization.
In his game-changing book The Gutenberg Galaxy: The Making of Typographic Man, which we don’t have space be give justice here, McLuhan studies the way technology for documenting and communicating has influenced human cognition and societal structures throughout history from pre-alphabetic human tribes to the electronic age. He argues that electronic media, through its instantaneous movement of information across the world, has shrunk the globe, bringing all matters of social and political function under our awareness and so heightening our sense of responsibility. Electronic media has rendered the world into a “global village”, argues McLuhan, and the next medium –which we can say today is digital media- would continue that process by extending our consciousness across the globe.
McLuhan’s logic works well in describing our world today; we are not only conscious of events happening across the world, we react to them as well. When natural disaster strikes a country, world-wide relief efforts respond. When human rights are violated by a state, demonstrations are held at that state’s embassies in other countries. And when an internet craze comes along, the world Harlem shakes, then looks back in shame wondering where it all went wrong. In such a world, the horrors of a world war and mass genocide cannot go without resistance. The world becomes a tribe beating to the same drum, and although disagreements and conflicts exist, the disassociation, alienation, fear and hatred of people from other countries needed for a full-out global war cannot foster in a world interconnected by the internet. This change is captured wonderfully in this video by Ronny Edry who started the We Love You – Israel & Iran campaign back in 2012.
At this point we can see what appears as a trajectory from print media to digital media. Print media consolidated geographic areas into imagined communities via language and by fostering a perception of simultaneous movement through linear time. Digital media shrinks and consolidates even larger geographic areas, arguably the whole world, via the language of the internet, simultaneity of online activity, and instantaneous sharing of information and context. The internet, however, goes beyond this. The reason McLuhan argues that new currents are underway is because he holds that electronic and digital media stem off from the trajectory initiated by print media and the book. This new direction is the major development challenging nationalism.
The wide-spread use of the book as a medium following the rise of the printing press, argues McLuhan, shaped human cognition into a visual-linear mode of thinking. Ideas and stories were communicated line by line. This had the effect of externalizing thought into a visible organized structure ordered by grammatical rules and conventions, thus making thought and cognition more objectively critiqueable, calculable, and logically linear. By emphasizing the visual and calculable, the sensuous and emotional become under emphasized and discredited as subjective. Human cognition becomes mechanical. Thus, McLuhan explains, print technology made possible the trends of the modern period such as capitalism, democracy, individualism, and, surprise surprise, nationalism. Thus, the development of linearity in human experience and perception which we have been finding recurrent in our examination so far –linearity in relationships, in time, and now in cognition- and which made possible the notions of competition, progress, and the nation, can be traced back to the proliferation of the printing press. And it is this linearity, McLuhan contends, which is being challenged by the proliferation of electronic and digital media.
The wide-spread use of electronic media as a medium induces a step away from linear cognition. The constant bombardment of stimuli we are exposed to whether on our computers, phones, or tablets not only bring about a revitalizing of the audio-oral but also train us in web-like cognition with multi-tasking awareness. While adults criticize youngsters today for short-attention spans and lack of concentrated focus, what goes unnoticed are youngsters capacities to carry out several activities at once –chatting on several social services, listening to playlists, downloading content, working on a school assignment, following the latest tweets, and texting from their phones. Whereas the book required the reader to focus her attention from line to line, the internet requires the user to branch out her concentration like an inter-linked web.
The internet, thus, challenges nationalism not just by expanding communities beyond borders, but –just as the shift from religious community to the nation involved a metaphysical shift- by shifting experience to non-linearity. As linearity is undermined so are notions of nationalism, national or ideological historical progress, and ultimately the notion of a global war as socio-politico-economic advancement.
.
Looking at memes, we can see that rather than continuing the practices initiated by the novel and newspaper, the meme is actually a social practice of non-linearity. Memes disrupt linear writing, with the structure of the superimposed text dictated and interrupted by images and not solely grammatical rules. The meaning of the meme is not contained within itself but comes from being inter-weaved with other memes, other people, and other events. And, most importantly, the meme is not a silent ceremony, but one which involves millions of users branching out and making themselves heard -voting, sharing, linking, posting, and commenting- through a giant web connecting them to people beyond borders and across the world. That is how memes could help prevent World War 3, or at least that’s what you can tell yourself next time you’re avoiding work. You’re welcome.
What would a world, then, shaped by non-linearity look like? How will the next imagined community, perhaps a global community, be structured? What kind of purpose or meaning can communities attain from a non-linear outlook? This blog has been a lengthy one, and there are a lot of points to talk about. Shrink the world further by letting us all hear what you think @catch21p or in the comments section below.
About the Author
Mark Bou Mansour has studied critical political theory and philosophy over the course of his undergrad and Master’s program, effectively turning his brain into mush. He now finds everyday things utterly fascinating and everywhere he looks he sees grand historical forces at play contingently shaping our worlds. Recently, he has taken a liking to shiny things like smartphones, computers, and web 2.0.
Paul Brand, Catch21 Ambassador
“We know that young people have a view, but too often they don’t have a voice. Catch21’s work is vital in encouraging 18-24 year-olds to participate in the political debate so that British democracy really does mean rule by the people – all people.”
Catch21 on flickr
Catch21 believes in democracy. The problem as we see it is that for democracy to function properly, everyone needs to have the same level of information. However due to inability, apathy and inaccessibility this is not always the case. Our aim is to remedy these factors by providing the access and information to participate and by empowering young people to make them feel a part of their communities. | |
Q:
Python3 Make a list that increments for a certain amount, decrements for a certain amount
if I have the following:
[0, 1, 2, 3, 4, 5, 6...] how can I reorder the list (actually make a new copy of the list) and then fill it as such:
[0, 1, 2, 3, 4, 5, 10, 9, 8, 7, 6, 11, 12, 13...]
I.e. Every five iterations, the list starts to decrement, or increment. The reason I want to do this is that I have a list of objects, and I want to fill a new list with the objects in a different order.
One technique I tried is:
copied_icons = [{key:'Object1'}, {key:'Object2'}, {key:'Object3'}...]
reversed_copied_icons = copied_icons[::-1]
left_to_right = []
for h in range(17):
left_to_right.append(copied_icons[h])
for j in range(18, 35):
left_to_right.append(reversed_copied_icons[j])
for k in range(36, 53):
left_to_right.append(copied_icons[k])
for l in range(54, 71):
left_to_right.append(reversed_copied_icons[l])
But for some reason this returns the list out of order and duplicates some of the objects. I am wondering if there is a simpler way to alternating incrementing and decrementing while filling my list.
A:
There are two problems with your approach:
You are reversing the entire list, not just that slice. Let's say the list is [1,2,3,4], and we want to reverse the second half, i.e. get [1,2,4,3]; with your approach, you would take the third and fourth element from the reversed list, [4,3,2,1], and end up with [1,2,2,1]
The to-index in a range is exclusive, thus by using range(17) and then range(18,35) and so forth, you are missing out on the elements at index 17, 35, and 53
You can use a loop for the different parts to be reversed, and then replace that slice of the list with the same slice in reverse order.
lst = list(range(20))
for start in range(5, len(lst), 10):
lst[start:start+5] = lst[start+4:start-1:-1]
Or this way, as pointed out in comments, which also gets rid of those nasty off-by-one indices:
for start in range(5, len(lst), 10):
lst[start:start+5] = reversed(lst[start:start+5])
Afterwards, lst is [0, 1, 2, 3, 4, 9, 8, 7, 6, 5, 10, 11, 12, 13, 14, 19, 18, 17, 16, 15].
Or, in case the intervals to be reversed are irregular (as it seems to be in your question):
reverse = [(3, 7), (12,17)]
for start, end in reverse:
lst[start:end] = reversed(lst[start:end])
| |
: 26 positive tests in latest results
Data released Wednesday by the NFL and the NFL Players Association confirmed the rise in the number of COVID-19 cases in the league.
In its weekly release of test results, the league said there were 26 positives among the 37,002 tests administered to 7,981 players and team personnel. The positives were attributed to 11 players and 15 team personnel.
The tests were performed Sept. 27-Oct. 3.
Quarterback Cam Newton of the New England Patriots was among the players to test positive in that span. The Titans' game against the Pittsburgh Steelers was moved from Week 4 to Oct. 25 because of an outbreak in Tennessee, which now has more than two dozen confirmed cases.
In the previous week's testing, two players and four staff members were positive among the 36,666 tests administered.
Since monitoring began on Aug. 1, the NFL has administered more than 370,000 tests with 31 players and 53 other personnel testing positive.
On Wednesday, Newton shared a reminder about the virus on social media.
Commissioner Roger Goodell's office issued a two-page memo detailing changes to COVID-19 protocols earlier this week. Changes include a longer onboarding process for free agent tryouts, bans on gatherings outside the club facility, limitations on the number of tryouts permitted per week, and the implementation of a league-wide video-monitoring system to ensure compliance with the protocols -- "particularly the mandate that all staff and players wear PPE while in a club facility and on travel at all times." | |
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About PQDT Open Search
A reawakening is simultaneously occurring in two traditionally opposing camps of moral and ethical theory: moral perfectionism and the pragmatic ethical theory of John Dewey. Moral perfectionism puts forth an objective theory of the good human life and subsequently bases moral claims upon this account Perfectionist theories posit a fixed end or ideal (or, if a series of them, usually in some kind of teleological order) whose attainment always ought to be sought. These ideal ends or objective goods exist outside of the immediate and specific moral problem at hand and guide ethical action by providing rules of conduct. Dewey's theory rejects the existence of any such objective and inherently valuable ends to ethical action, instead putting forth a normative theory centered on his novel conception of growth. Deweyan goals or ends constantly evolve in light of new discoveries and experiences and are subjected to collective experiential verification within specific social settings. Instead of steadfast rules, there are principles that are rooted in experience and which evolve in light of new evidence; instead of ideal universal ends and objective goods, there are ends-in-view that serve as tools in the resolution of moral problems.
Both perfectionism and Deweyan ethics, having previously fallen out of favor for being associated with questionable or deficient interpretations and doctrines, are being reconsidered with growing interest. Additionally, attempts to bridge the two camps are emerging, and these usually claim that Dewey's ethics espouse perfectionist tenets. This dissertation stands at the intersection of these emerging camps: my goal is to offer a contribution that clears the ground by clarifying what, first, Dewey has to say about perfectionism and, second, how this relates to the cogency of his ethical theory as well as the validity of contemporary attempts to cast Dewey as a perfectionist. In the course of pursuing this program, I demonstrate how Dewey thoroughly rejects traditional "thick" versions of perfectionism that prescribe relatively restrictive criteria of human good. Upon scrutinizing Dewey's theory from within the correct framework—his own reconstructed one—I reveal how it is, indeed, coherent although it may not be sufficiently adequate. Additionally, in light of the exegesis clarifying his ethical theory, I show the recent reinterpretations of Dewey as a perfectionist are flawed and inadequate. However, new perfectionist theories currently being developed stipulate more flexible (or "thin") criteria, taking into account situational variation and recognize the importance of contingent experiences. It is possible this "thin" species of perfectionism does not succumb to Dewey's critique of perfectionism and that Dewey's own ethical theory and his notion of education as growth can be reinterpreted as incorporating thin perfectionist premises. The groundwork provided by this dissertation allows for further exploration of this open question.
|Advisor:||Phillips, Denis C.|
|Commitee:|
|School:||Stanford University|
|School Location:||United States -- California|
|Source:||DAI-A 71/01, Dissertation Abstracts International|
|Source Type:||DISSERTATION|
|Subjects:||Ethics, Philosophy, Education philosophy|
|Keywords:||Dewey, John, Ethical theory, Experimental life, Moral education, Perfectionism, Philosophy of education|
|Publication Number:||3395870|
|ISBN:||9781109583601|
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the next 'Rhythm of Life' Drum Circle will be at this event, from 5-7pm!!
Drum circles are an ancient form of experiential therapy that promote greater mental, physical and spiritual health. Recent research indicates that drumming accelerates physical healing, boosts the immune system and produces feelings of well-being, a release of emotional trauma and reintegration of self. | https://recovery.wvu.edu/event-calendar/special-events |
These are the documents on which the Union or the central government collects a stamp duty. In addition, national governments may also impose taxes on certain documents. 1. In the case of (f), the extra-account obligation is granted when carrying out the transport assets. To validate such a real estate transaction, the buyer must pay stamp duty, as proof of the purchase has been provided. Stamp duty is therefore the tax paid by the state at the time of the real estate transaction and has the transfer certificate properly kept in court. Q10. What are the provisions/articles of the Indian Stamp Act, 1899 attracts stamp duty in the state of Maharashtra? The registration fee for a rental contract in Maharashtra, depends on the location of the rental. The registration fee is Rs 1,000 if the property is below an urban corporate area and it is 500 aff. if this is the case in a rural area. In the absence of a contrary agreement, the tenant bears the costs of stamp duty and registration.
1. A sale agreement relating to the transfer of ownership is considered a “transportation” and is marked accordingly. However, the tax paid is set at the time of the transport. Under the Indian stamp law and most state stamp duty laws, stamp duty instruments are inadmissible evidence in the event that no appropriate stamp duty has been paid. Section 35 of the Indian Stamp Act deals with the consequences of not stamping documents. It states that, for Maharashtra, the law has been strengthened and that, in accordance with the provisions of Section 55 of the Maharashtra Rent Control Act of 1999, any lease or leave and licence agreement must be written down, and the same must be registered in a mandatory manner, regardless of the length of the lease. Stamp duty must be paid on instruments and not on transactions. The definition of the instrument is very broad. Also help me with the amount of the fine, if we re-develop documents in a month, since the tax had not been paid before, we have to pay 100% fee, 200% fee, or how much? Use: Payment of stamp duty for transactions with civil and criminal courts.
Q29. What happens if the instrument is executed on stamps that do not bear the name of one of the performers? As far as the duty of the state is concerned, it generally varies from state to state. Nevertheless, there is a general pattern that is followed. Let`s take a look, for example, at the stamp duty imposed by the Karnataka government. Apart from the documents mentioned above, the Karnataka government collects stamp duty: See it is not mandatory to pay stamp duty and the francization of the master service contract. However, improperly stamped instruments may be admitted as evidence in the event of payment of the applicable tax and the prescribed sanction. 4.4 In addition, s.14 prohibits the letter of a second taxable instrument on a stamp on which a taxable instrument has already been written. With respect to your second question, you may need to examine the corresponding stamp law to determine whether a specific stamp duty should be paid or whether Rs. 100 general stamp paper would be sufficient. General stamp paper of such an amount as Rs. 100 (which may vary from state to state) is also essentially a stamp duty, but it is a standard rate for all different agreements for which there is no specific stamp duty. Therefore, it is advisable to consider certain clauses in the timetable of the stamp law in question, if there is mention of a specific stamp duty for certain instruments.
Anyone who, by law or with the permission of the party, gets evidence, and anyone for? the public office before which each instrument accused of stamp duty is produced or enters into the performance of its duties. | https://www.chocolatetreasuresnj.com/stamp-duty-on-service-agreement-in-mumbai/ |
Admitting Unit.
During my various experiences, I have encountered patients suffering from a wide range of
respiratory conditions, including bronchogenic carcinoma, interstitial lung disease, pleural
effusion, Chronic Obstructive Pulmonary Disease, bronchiectasis and respiratory failure. I have
successfully diagnosed respiratory conditions and provided appropriate treatments. As well as
dealing with specific respiratory cases during my dedicated respiratory medicine rotations, I have
needed to maintain an awareness of respiratory conditions to allow me to identify and treat those
which may emerge during my other rotations such as emergency and general medicine. I have
identified respiratory conditions through physical exam, taking of patient and family health
history, and tests such as pulmonary function tests.
Adding to my exposure to respiratory medicine, my diverse and rigorous training and experience
in other aspects of medicine add to my ability to provide safe care of patients. In 2010 I had the
opportunity to commence working as a medical officer at the La Trobe Regional Hospital. In this
hospital I successfully completed rotations in emergency, psychiatry and surgical specialties, and
rehabilitation medicine. To expand my horizons and learning opportunities, I moved to Frankston
Hospital as a senior medical officer in 2011. As a senior medical officer at this hospital, I was able
to accumulate valuable clinical experience in a wide variety of fields including general medicine,
emergency medicine, respiratory medicine, oncology, cardiology and neurology.
For the past two years I have been undertaking Basic Physician Training at Sir Charles Gardiner
Hospital, completing training in renal medicine, neurology, general medicine, geriatric medicine,
medical oncology, rehabilitation medicine and the intensive care unit. The extensive and thorough
training and experience I have received throughout my career to date, ensures that I am
wellequipped to provide care as a Registrar in respiratory medicine.
Demonstrated ability to provide medical education, teaching, supervision, training and support
to resident medical officers and interns
As a Registrar I have gained a substantial amount of experience in the supervision of interns and
resident medical officers in a hospital setting. During ward rounds, I am accompanied by interns
and residents and provide information and advice and allow for the extension of knowledge by
encouraging questions and asking for the opinions of those accompanying me. I provide training
through demonstrating medical procedures and skills, such as detection of clinical signs and taking
of history from patients. Maintaining a flow of communication and discussion through ward
rounds, I discuss the actions I am undertaking and the reasons for my actions. This demonstration
allows interns and residents to then gain confidence in performing procedures themselves for the
first time.
I undertake hospital teaching through case presentations and make myself available for questions,
discussion and advice. This extends to making myself available for after-hours calls to answer
questions over the phone. With the belief that it is important for the development of knowledge
to be able to gain experience in a supportive environment, I make efforts where appropriate to
allow interns and resident medical officers to undertake new procedures and practice new skills
under my guidance. While new skills are being developed I provide supervision and guidance, and
make myself available afterwards for feedback and discussion. Through providing positive
feedback and constructive advice for improvements as appropriate, I encourage the development
of high standards of ability in those whom I train.
Maintenance of a professional and approachable demeanour provides interns and residents with
the confidence to approach me with questions or requests for advice. By taking the time to
demonstrate and discuss techniques, conditions and diagnoses, I contribute to the development of
knowledge. I ensure that my communication with interns and residents is clear and concise, in
order to accurately provide information that will be useful to them in their future medical practice.
As a doctor who reads widely to extend and maintain currency of my own knowledge, I enjoy the
opportunity to share my learning for the benefit of others. Through discussion of current research
or new published material, new knowledge is gained.
Demonstrated verbal and written communication skills and interpersonal skills to effectively
interact with patients, their families and staff at all levels
A confident and clear communicator, I am equally comfortable conversing through written or
verbal means. While most of my interaction in my role as a doctor is in person, I possess a
demonstrated ability to communicate clearly across electronic and print forms, on the telephone
and in face-to-face environments. As an effective communicator, I am able to adapt my
communication style to suit my audience.
In my personal communications I possess a professional and amenable interpersonal style and am
able to communicate professionally and appropriately with all levels of staff, patients and their
families. In communicating with patients, I make efforts to place myself in the shoes of the patient
and understand the hospital experience from their perspective. In this way, I can empathise with
their concerns and more effectively explain their diagnosis, proposed treatments and prognosis.
Through understanding of the patient’s needs and responses, I am able to build rapport with the
patient and their family. This rapport encourages the open exchange of information and allows me
to provide reassurance and comfort to patients and families who are upset by the hospital
experience and hence resistant to treatment that would be in the best interests of the patient.
My interactions with staff see me maintain professional courtesy with all hospital staff and
communicate clearly and concisely with appropriate respect for the position held and the
professional knowledge and expertise of the staff member. I am able to build good relationships
with hospital staff, which serve to enhance the care of patients.
With English as my second language, I undertook the International English Language Testing
System (IELTS) testing in 2009, the same year I arrived in Australia. Obtaining scores of 7 and 7.5 in
each of the categories (listening, reading, writing and speaking), my band score falls in the
category described as “generally handles complex language well and understands detailed
reasoning’. I have continued to develop and refine my English language fluency over the past six
years in Australia and continue to seize opportunities such as the chance to make case
presentations to continue to extend the effectiveness of my communication.
Demonstrated organisational and time management skills to provide safe, timely patient
centred care
An effective planner, I am able to manage my shifts and my caseloads in a manner which allows
optimum care to be delivered to my patients. Meticulous in my personal and professional habits, I
ensure that attention is paid to the detail in my work. This ensures that errors are not made and
patient care is not compromised through haste or inattention. My attention to patient needs
supports active involvement of patients and their families in decision-making about individual
options for treatment.
Through planning of my day I am able to maintain effective care of a high volume of patients while
maintaining the ability to attend to constantly evolving priorities. Even in high-pressure, rapidly
evolving environments such as emergency medicine, I have found that my organisational and
planning skills have provided me with the ability to attend to urgent matters within appropriate
timeframes, without being distracted by other less urgent demands. By prioritising and
maintaining a clear understanding of the relative urgency of competing demands on my attention,
I am able to ensure that optimum patient care is not compromised by a high workload.
I am conscious that patient experience of hospitals can at times be frustrating, and I ensure that I
communicate clearly and regularly with patients to ensure that they are informed of the next
steps in their treatment and when they can expect those steps to occur. By communicating well
and ensuring that the patient is notified of any delays, I contribute to the patient’s experience of
feeling that they have received appropriate and timely care.
Priding myself on punctuality and reliability, I keep commitments I have made to my colleagues
and complete actions by the agreed time. I maintain open lines of communication with fellow
health professionals on my patient’s care team, ensuring that others know what they can expect
of me and when I will be able to deliver treatment or results. As a single person, I commit my time
to my profession and furthering my medical knowledge, experience and skill. I am more than
willing to undertake overtime and participate in on-call roster as these assignments provide me
with additional opportunities to develop as a physician.
Demonstrated participation in continuing medical education activities including regular
performance reviews to maintain and upgrade knowledge & skills
Demonstrating my commitment to my profession and furthering my medical knowledge, I
dedicate substantial amounts of my personal time to reading new studies and journal articles in
order to broaden and deepen my knowledge of medical practice and contemporary methods.
Wherever possible I attend teaching sessions, lectures and stimulation courses. I attend weekly
grand rounds and participate in presenting the medical problems and treatment of a particular
patient to an audience consisting of doctors, residents and medical students.
At the commencement and conclusion of each rotation, I conduct a learning needs analysis to
ensure that I am obtaining the necessary knowledge and competency through my experience in
each particular field of medicine. I also participate in mini-Clinical Evaluation Exercise cases to
assess my learning and competency.
Demonstrated ability to monitor and review their clinical effectiveness, note their limitations
and put in place strategies to ensure safe patient care
It is a habit, in my professional practice, to continually assess my own performance and identify
the areas where I could improve my knowledge and skill or seek opportunities to gain further
experience. Whilst I have a strong interest and thorough theoretical knowledge of common
respiratory medicine conditions, I recognise that my direct experience in this field is still
developing. To this end, I would ensure that in practicing as a Registrar in the field of respiratory
medicine, that I did not allow over-confidence to compromise patient care.
By drawing on the knowledge, experience and advice of professionals around me, I would aim to
extend my own knowledge in any area where I identified weakness. I am aware for example that I
do not possess particular expertise in interpreting CT scan images; for this reason I would be likely
to draw on the expertise of the radiology department in reading CT results. Likewise, I would
potentially request supervision the next time or two I undertook a pleural tap procedure, as this is
something I have not done in some time. Through recognising areas such as these where my
knowledge is still developing, and putting in to place strategies to ensure that I can effectively gain
experience without compromising patient safety, I demonstrate awareness of my own ability and
the capacity to monitor my own effectiveness.
I am dedicated to continuous improvement in the quality of care that I provide to my patients. In
this process, it is important to have a specific improvement program with a well-defined objective.
To this end, I set myself concrete goals of extending my knowledge in particular areas where I feel
my skills could be improved. I then seek opportunities to learn through observation, attending
lectures or seminars, reading articles or initiating discussions with other physicians. Through taking
as many opportunities as I can to observe and then repeatedly undertake a procedure, I improve
my level of expertise and ability, rather than allowing myself to develop areas of skill shortfall. In
this way I continue on my journey to become a well-rounded physician with depth and breadth of
skill and knowledge. | https://selectioncriteriaexamples.com.au/registrar-trainee-respiratory-medicine-2/ |
Diabetic neuropathy is a complication of diabetes that causes nerve damage. Over time, high levels of glucose and fats in the blood can damage nerve coverings, as well as blood vessels that carry oxygen to nerves. Diabetic neuropathy can happen anywhere in your body. Types of diabetic neuropathy include:
- Autonomic neuropathy, which affects nerves that control internal organs
- Focal neuropathy, which typically affects a single nerve in the head, hand, torso or leg
- Peripheral neuropathy, which affects the legs and feet and sometimes the arms and hands. This is the most common form – as many as half of people with diabetes experience peripheral neuropathy.
- Proximal neuropathy, a rare type that affects one side of the body in the hip, thigh or buttock
Diabetic neuropathy may cause symptoms that include:
- Numbness, burning or tingling, weakness and shooting pains in hands, feet or legs
- Pain when walking
- Leg weakness
- Loss of muscle strength and muscle tone
- Loss of reflexes
- Problems with sexual function
- Carpal tunnel syndrome, caused by nerve compression at the wrist
- Dizziness when changing positions quickly
- Constipation and diarrhea
- Nausea and vomiting
- Problems swallowing
- Problems sensing pain or temperature changes in parts of the body
- Vision issues, especially problems adjusting to changing light conditions
- Sudden, sometimes severe pain in a hip, buttock or thigh
- Swollen feet
- Urinary problems
Over time, high levels of blood glucose and fats, such as triglycerides, can damage nerve coverings or the blood vessels that carry oxygen to nerves.
You can help prevent diabetic neuropathy by properly managing your diabetes. This includes:
- Keeping your blood glucose levels in your target range
- Monitoring your blood pressure and cholesterol
- Taking good care of your feet and checking them daily for problems
- Seeing your health care provider for an exam at least once a year to check for neuropathy in feet and other areas of your body
Diabetic neuropathy can lead to serious health problems if left untreated.
Complications of diabetic neuropathy include: | https://www.ukhealthcare.uky.edu/barnstable-brown-diabetes-center/diabetes-complications/diabetic-neuropathy |
sought to bury him in Springfield.
The bodies of Lincoln and his son Willie were first placed in the cemetery's receiving vault, because the tomb was not completed until 1874. Designed by Larkin Mead of Vermont, the monument features a 117-foot granite shaft and famous statuary.
Dedicated October 15, 1874
Shortly after Lincoln's assassination, a special public meeting met to determine the most appropriate method of memorializing their late president. A special committee from this meeting resolved itself into a National Lincoln Monument Association and was headed by Governor Richard Oglesby. Several prominent Springfield men, most personal friends of Abraham Lincoln, comprised the group, among them John T. Stuart, his first law partner, and O. M. Hatch, a friend and ex-Illinois secretary of state. The Association planned and began construction on a special vault for Lincoln located on the grounds chosen for the new statehouse, but Mrs. Lincoln objected, insisting that her husband rest forever in the city's rural Oak Ridge Cemetery. Honoring the first lady's wishes, the Association arranged for the president and his son Willie, who died in 1863 and traveled with his father to Springfield, to lay in the Cemetery's public receiving vault. In the meantime, workers began constructing a temporary vault where the president would lay until the completion of his National Monument and Tomb. On December 21, 1865, after verifying the remains, officials moved the president, Willie, and another son, Eddie, into this temporary vault.
In 1868, after a major fundraising campaign, the Association sent out a call for designs. They received thirty-seven designs, choosing one by Larkin G. Mead of Vermont. Mead, a sculptor, contracted to produced the exterior statuary while local contractor W. D. Richardson agreed to construct the monument itself.
Work proceeded on schedule, and by 1871, although not complete, the tomb was ready to receive its first interment, only it was not that of the president. Lincoln's son, Thomas ("Tad"), died of fever after returning from Europe. On July 17, 1871, Thomas became the first Lincoln to occupy the tomb. On September 19 of the same year, his father and two brothers joined him. At that time, those responsible decided to remove Lincoln's body from the wooden coffin where it lay and seal it in a metallic burial case.
The Association continued its efforts to raise the $171,000 necessary to complete the tomb and monument. The total cost covered the construction of the monument, including the foundation, base, terrace, and obelisk, as well as the statuary of Lincoln and the four armed forces groups representing the infantry, cavalry, artillery, and the navy that would adorn the obelisk's pedestal. Public enthusiasm and support was high and the Association was able to secure subscriptions to meet financial needs. Additionally, they secured the support of prominent individuals and families from four cities to underwrite the cost of the pedestal statuary. Consequently, the infantry group was a gift from the city of Chicago, the naval group was furnished by New York City, Boston donated the cavalry statue, while Philadelphia provided funds to finish the artillery group.
Lincoln's sarcophagus in the monument's burial chamber lies below the floor in a steel reinforced concrete crypt. It has remained there since 1901, after a major reconstruction project. The flags surrounding the marker represent four states that were homes to Lincoln's ancestors (Massachusetts, New Jersey, Pennsylvania, and Virginia) and the three states where Lincoln lived (Kentucky, Indiana, and Illinois). The U.S. and presidential flags are also included. Secretary of War Edwin M. Stanton's famous words, "Now he belongs to the ages," adorns the marble above and behind his marker. Mary Todd Lincoln and three of the Lincolns' sons are buried in a family crypt located behind visitors as they face Lincoln's marker. Their names are carved into the tomb's wall.
buried ten feet under this marker
his wife, Mary Todd Lincoln
is interned in the opposite wall
their young son
other children
On October 9, 1874, officials removed Lincoln from his tomb, verified the remains once again, put his body in a lead-lined, red cedar coffin, and placed him in a marble sarcophagus in preparations for the dedication of the Monument six days later. On that day, amidst an enormous crowd, including veterans of the Army of the Tennessee attending its Eighth Annual Reunion, the Monument was dedicated with speeches, music, and the unveiling of the Lincoln statue on the pedestal. At the end of his address, the Honorable Jesse K. Dubois, the Association's Vice President, hoped that "There may [he] rest in peace." But he wouldn't for some time.
Two years after the dedication, Lincoln's body escaped a failed attempt by a counterfeiting ring to steal his body and hold it for a ransom of $200,000 and the freedom of the gang's imprisoned master engraver. In all, Lincoln's coffin has been moved 17 times, mainly due to reconstruction, and has been opened five times. The last time for both was September 26, 1901, when officials verified that the remains were Lincoln's and then set his coffin in a concrete crypt beneath the monument's floor surrounded by reinforced steel, in part to prevent further attempts at desecration. Despite additional reconstructions, Lincoln's remains have rested in peace since 1901. Thousands continue to journey to the tomb each year to pay their respects to one of the country's most revered leaders. It seems that the National Lincoln Monument Association completed its task of erecting a tribute that conveys the country's estimate placed upon his life, virtues, and public services. | http://www.galenfrysinger.com/springfield_lincolns_tomb.htm |
Tyson R, Graham JP, Bermingham E, et al.
Vet Radiol Ultrasound 2005;46:33-38.
The purpose of this study was to describe normal feline hypophyseal mensuration and contrast enhancement characteristics using dynamic computed tomography (CT) imaging. An intravenous bolus of an ionic iodinated contrast medium was administered to eight cats using a pressure injector while dynamic CT images were obtained every 5 s for five cats and every 7 s for three cats for a total imaging time of 5 min. Each pituitary was measured at its maximum height and width on the peak contrast medium enhancement image. A hand-drawn region of interest was placed around each hypophysis cerebri and time attenuation curves were generated. The specific enhancement pattern of the hypophysis cerebri for each cat was recorded. The mean width and height of the hypophysis cerebri was 5.2 +/- 0.4 (average +/- SD) mm and 3.1 +/- 0.3 mm, respectively. The mean time to maximum contrast enhancement was 28.6 +/- 14.8 s (range 14-50 s) from the onset of contrast medium injection. Four cats had initial dorsal and peripheral contrast enhancement patterns of the hypophysis cerebri, while four cats had an initial central contrast medium enhancement pattern. The hypophysis cerebri had a homogenous appearance in all cats, 28-50 s after contrast medium injection. The average (+/- SD) clearance half-time was 292 (+/- 87) s. Normal hypophysis cerebri mensuration and contrast medium enhancement characteristics will help in clinical evaluation of the feline hypophysis cerebri. | https://avmi.net/information/ct-concepts/dynamic-computed-tomography-of-the-normal-feline-hypophysis-cerebri-glandula-pituitaria/ |
Found this vintage video on You Tube. Thirty-six years ago, when I got my first van, a Ford Econoline, this was the type of lift the converter put in. It was called a Golden Boy. The conversion included lowering the floor behind the front seats a bit so I didn't struggles ducking my head out the door like the fellow in the video. Thirty-six years ago was before the advent of the "compact car" parking space. The van could park in a full sized parking space with a car next to it and still swivel out with some room to spare between the lift and the car beside the van. Thirty-six years ago, not many parking garages could accommodate the height of the full sized van. Six feet, eight inches was about the best accommodation you could get. Now most garages have at least one section that will accommodate over seven feet. But, I find being restricted to one section of a garage is inconvenient, if all the handicapped spaces there are occupied. I prefer the mini van that allows access to other parts of a garage with more handicapped spaces available.
I don't know why the swivel lift was discontinued. | http://sci.rutgers.edu/forum/showthread.php?264260-van-verse-car&s=6ff23c7f7153cd08092afcdead229cad&p=1875650 |
An editorial by Miranda Robertson in the Journal of Biology ("Biologists who count", 8(34), 2009), starts this way:
The importance of mathematics in biology is a matter of perennial debate. The squabbles of early 20th century geneticists on the value of mathematics to the study of evolution have recently been revisited in Journal of Biology [Crow, "Mayr, mathematics and the study of evolution", JBiol 8(13) 2009], and the 21st century has seen an explosion of information from various -omics and imaging techniques that has provided fresh impetus to the arguments urging the need for mathematical competence in the life sciences [Bialek and Botstein, "Introductory science and mathematics education for 21st-century biologists", Science 2004, 303:788-790]. While there can be no question about the contribution of mathematics to many fields in biology, there is a curious tendency on the part of numerate biologists (often immigrants from the physical sciences) to insist that it is an essential part of the equipment of a biologist and none should be without it. This seems, on the evidence, extreme.
Robertson concludes that
There seems no need for the snobbery (it is said) of the highly quantitative founding biologists at the Cold Spring Harbor Laboratories, in whose early history ex-physicists played a crucial part, and who are alleged to have referred to their nearby colleagues at Woods Hole as biologists 'who don't count'.
There's an analogous set of arguments in the areas of rational investigation related to language, of which the discipline known as "linguistics" is a regrettably small part. (I mean by this that the balkanization of the field is a Bad Thing, not that the work of psycholinguists, linguistic anthropologists, speech pathologists, speech technology researchers, etc., is inadequate or inferior. But that's a different discussion.)
The role of mathematics in the language sciences is made more complex by the variety of different sorts of mathematics that are relevant. In particular, some areas of language-related mathematics are traditionally approached in ways that may make counting (and other sorts of quantification) seem at least superficially irrelevant — these include especially proof theory, model theory, and formal language theory.
On the other hand, there are topics where models of measurements of physical quantities, or of sample proportions of qualitative alternatives, are essential. This is certainly true in my own area of phonetics, in sociolinguistics and psycholinguistics, and so on.
It's more controversial what sorts of mathematics, if any, ought to be involved in areas like historical linguistics, phonology, and syntax.
I don't have time this morning for a longer discussion, so I'll just baldly state my conclusions, and look forward to hearing the opinions of others.
First, attempts to devise and test formal models of language-related phenomena are often helpful, and sometimes essential, in framing theories clearly enough to be able to see what's right and what's wrong with them. And occasionally, such models lead to genuine insight.
Second, mastery of statistical techniques is useful in all forms of rational inquiry; and networked digital computers are increasing this usefulness in a major way, due to increased availability of data and increased ease of statistical investigation.
Third, mastery of language-relevant mathematics crucially includes knowing when a model is inappropriate, misleading or unnecessary. Yeats said this in a more pointed way, in his 1930 "Letter to Michael's Schoolmaster":
Teach him mathematics as thoroughly as his capacity permits. I know that Bertrand Russell must, seeing that he is such a featherhead, be wrong about everything, but as I have no mathematics I cannot prove it. I do not want my son to be as helpless.
I happen to think that Russell was, on the whole, righter than Yeats was; but it would be wrong for the argument to be won by default due to one side's technical incompetence. If you've ever worked in an interdisciplinary area where mathematical backgrounds are variable, you've probably seen attempts to win arguments by this sort of mathematical default — often promoted by people who don't really understand a technique or program that they've learned to use in a cookbook fashion.
(I certainly don't subscribe to the part of Yeats' letter that says "Don't teach him one word of science, he can get all he wants from the newspapers and in any case it is no job for a gentleman". Alas, many of those who write about science in the newspapers seem to have been educated in accordance with this prescription.)
Anyhow, my conclusion is that anyone interested in the rational investigation of language ought to learn at least a certain minimum amount of mathematics. | https://languagelog.ldc.upenn.edu/nll/?p=1461 |
According to new legal documents, obtained by The Blast, Depp filed a subpoena to Musk to produce and protect all items that are associated with Amber Heard.
The information listed in the documents to turn over include, “conversations, telephone calls, letters, emails, memoranda, reports, telegraphs, faxes, exhibits, drawings, text messages, and any other documents which confirm or relate to the written or verbal exchange.” also any electronically stored information including, “electronic, digital, or any other recorded material whatsoever, including but not limited to, any notes…videotapes, affidavits, statements, papers, files, forms, data, tapes, printouts, letters, reports, communications, contracts, agreements, telegrams, records, financial records, applications, correspondence, diaries, calendars.”
The request specifically asks for communications around the May 21, 2016, domestic violence incident, where Amber claims Johnny Depp assaulted her in their home.
“All communications relating to allegations of physical abuse or domestic violence committed by Mr. Depp or Ms. Heard,” it reads.
As we’ve reported, several witnesses have testified to seeing Musk enter Depp and Heard’s home around the time of the incident. | https://johnnydepp.blogabode.com/2019/10/09/elon-musk-has-officially-been-subpoenaed-in-by/ |
For impact investors, it’s not enough just to earn a profit.
ETFs, which trade like stocks, and are subject to investment risk.
Equity securities, which may fluctuate in value in response to the activities of individual companies and general market and economic conditions.
Non-diversified mutual funds that focus on a relatively small number of issuers, which tend to be more volatile than diversified funds and the market as a whole.
Fixed-income products that include corporate bonds, municipal bonds, and other types of debt securities.
Companies with large market capitalizations, which can go in and out of favor based on market and economic conditions.
Through these and other types of marketable securities, we strive to help impact investors make good choices with their investment capital. | https://globaladvisers.com/what-we-do/sustainable-investing/impact-investing/ |
Visioning, Project Start Ups, Problem Solving and Team Building
I have over 25 years of experience with a wide range of groups with differing agenda and seniority. Part of this work has covered Visioning and Strategy development, as well as working with Director/Senior Management teams looking to develop their leadership style, or simply finding ways of working together. This type of work is especially useful to Project Managers and their teams in order to shape project plans and deliverables, as well as building the actual team to deliver them.
My previous work for clients has covered:
- Visioning, Strategy Development
- Problem Solving sessions.
- Team building events
- Conferences
- Focus groups
- Soliciting staff feedback
- Project/Programme start ups
- Project/Programme reviews
It is important for me as a Facilitator to:
- Be clear on what outcome the client wants from the event. This may take several meetings to establish, as sometimes the client needs to work out their needs first.
- Have a clear design to achieve those outcomes.
- Be sensitive to the energy and mood in the room.
- Respond to what is happening in the room and move away from the event design if necessary to achieve the client outcomes (liaising with the client as I do it).
- Be flexible and creative.
- Be able to offer the group a range of styles to help them achieve their goals (eg being Directive or Facilitative) as the situation demands.
- Be able to offer the group different techniques and ideas to challenge or stimulate their thinking.
- Be clear on why I am making an intervention and what I want to achieve with it.
- Review the workshop with participants and co-facilitators to learn and sharpen my experience and practice.
Before the workshop I will:
- Help you to be clear about the outcomes you want from the session.
- Design the session around your specific needs and complete any required preparation.
- If required, conduct interviews with the participants to explore their wants from the session.
During the workshop I will help you to:
- Stay focused on the key issues and the desired outcomes from the session.
- Create a safe environment in which to exchange honest views and be creative.
- Fully engage the group and develop a common understanding of the issue and solution at hand.
- Identify how you will implement the results. | https://vision4change.com/workshop-facilitation/ |
BACKGROUND OF THE INVENTION
I. Field of the Invention
The present invention relates to obtaining and maintaining synchronization with an incoming signal prior to and during the transmission of data. More specifically, the present invention relates to recognizing that a pattern within an incoming demodulated electromagnetic signal has repeated itself (or that some mathematical relationship thereof has been received) a specified number of times, indicating that synchronization has been achieved and that transmitted data is forthcoming. The present invention also more specifically relates to maintaining synchronization with the incoming demodulated signal while data, which is imbedded within the signal, is received.
II. Related Art
The ability to transmit and receive information using a wireless electromagnetic carrier (at, e.g., radio or microwave frequencies) has been known for many decades. A somewhat more recent development, though, is the ability to receive and process binary data using digital technology. One common way that this works in a wireless environment is that an electromagnetic "pattern" is transmitted representing a binary piece of data (i.e., a binary 1 or 0). This pattern transmission may utilize amplitude, frequency or phase modulation.
The constituent pieces of the pattern themselves represent binary values. Thus, a pattern representing several binary values are used to represent a single binary value of data. For example, receipt of the pattern "10111011" may represent the receipt of "011 as data, while its inverse, 1101000100" may represent receipt of "1. " Data is conveyed in this way because of noise and interference associated with wireless transmissions. Otherwise, it would be difficult to determine whether the receipt of a single binary value was data or just noise.
Information on the electromagnetic carrier is thus received as a stream of representative binary 1's and 0's. Each binary 1 or 0 is known as a "chip." Thus, the data being transmitted is in the form of a pattern of chips. It is desirable that a pattern representing a binary piece of data comprise a significant number of chips to reduce the chance that unwanted noise in the electromagnetic information is mistaken for transmitted data.
Applications utilizing the transmission of digital information have become very popular in recent years. One such application which has been getting wide-spread attention is wireless local area networks (wireless LAN's). This technology allows computers to exchange information with each other by transmitting this information as described above. The obvious advantage that wireless LAN's have over other LAN's is that wires are not needed to connect the computers together.
Because the airwaves are already very crowded and heavily used by a variety of technologies, it is difficult to assign a usable band of frequencies for use by wireless LAN'S. Consequently, spread-spectrum technology is often utilized. This technology (which was originally developed for military applications) allows an electromagnetic transmission to be spread throughout many different frequencies which are already assigned to such technologies as radio or microwave devices. Since spread spectrum signals have a low-power spectral density, they generate less interference to other devices tuned to the same frequency.
Spread spectrum technology and wireless LAN's are particularly applicable to notebook and other small portable computers, as these computers are typically purchased for mobility. However, because of their smaller size and dependency on batteries for power, it is important to minimize the amount of circuitry used in receiving the different frequencies. In addition, because of the competitive nature of the computer industry, it is also important that costs associated with spread spectrum devices associated with wireless LAN's be kept to a minimum.
As indicated above, noise and interference are a problem whenever information is transmitted by electromagnetic means. Because of this problem, two important issues must be resolved for data to be successfully received. The first is that the received information needs to be initially recognized as valid information as opposed to noise. This recognition of valid information is known as "synchronization." During this initial synchronization the information acts as a header to indicate that data is about to be received. The second issue is that once synchronization has been obtained, it must be maintained while data is transmitted. This typically involves maintaining alignment of the received information with a reference signal.
Of relevance to both these issues are two general methodologies for determining whether synchronization has been obtained as is being maintained. The first is to analyze the incoming information on the modulated electromagnetic carrier for synchronization purposes prior to converting the information into digital form. The second is to first demodulate the information (i.e., remove the information from the electromagnetic carrier), convert it into digital form and then analyze the digitized information. The advantage of the latter methodology is that it produces a more cost-effective and compact design. Unless stated otherwise, this is the methodology assumed in use in the discussions below.
Regarding the first issue of obtaining synchronization, as indicated above the transmitting source typically transmits a synchronizing header comprising some pre-determined pattern. The received transmission is compared with a reference code (also comprising the pre- determined pattern). A match of the received transmission and reference code indicates that the header has been received and that data is about to be transmitted. The longer the pattern, the less likely it will be mistaken for noise or interference.
Although a longer pattern is obviously preferable, the problem is that the number of components (and thus the size of the circuit) required to detect synchronization increases nearly exponentially as the length of the pattern increases. When dealing with size, cost and power consumption constraints as discussed above, use of a long pattern can quickly become uneconomical.
Consequently, what is needed is a scheme to utilize an adequately long pattern to minimize false detection while also keeping the size of the synchronization detecting circuitry to a minimum. In addition, the circuitry needs to work in an environment where the incoming electromagnetic information has been demodulated first.
Once the demodulated information from the electromagnetic carrier (hereafter referred to as the "input signal") has been synchronized, then data is received. As indicated above, maintaining synchronization of the input signal as data is received is the second important issue which must be resolved to successfully receive the data.
Also as indicated above, typical schemes for conveying data transmit patterns where each pattern represents a 1 or a 0. The patterns used to transmit data are usually (but not always) longer than ones used for obtaining synchronization. However, the mere comparison of a pattern used to transmit data with a reference code in the manner described above is insufficient for maintaining synchronization. This is because fine adjustments are typically made to realign the input signal. This requires determining whether the input signal is being transmitted too quickly or too slowly, which is not deducible from just a pattern comparison.
Schemes which utilize patterns of an input signal to maintain synchronization during data transmission consequently use a somewhat different approach from that used to initially obtain synchronization. One such scheme for maintaining data synchronization utilizes what is known as a delay lock loop. Although this scheme is conventionally used prior to demodulation, it is discussed below in conjunction with FIGS. 1A and 1B with regard to a binary post-demodulation environment.
Referring first to FIG. 1A, a diagram of a conventional delay lock loop is shown. In operation, an input register 102 receives an input signal via line 100. The input register 102 is divided into two sections (124 and 126), each of which contains a chip received from the input signal. The chips shown in input register 102 represent the chips that are in the input register 102 during a time period t.sub.c.
It is contemplated, time period t.sub.c is the time between when a single chip is first detected (in a reference code presumed in substantial alignment with the input signal as discussed below) and the time that the next chip is subsequently detected. Thus, as shown in FIG. 1A, in roughly the first half of time period t.sub.c, section 124 contains chip N while in the second half of that time period it contains chip N+1. Similarly, in the first half of time period t.sub.c section 126 contains chip N-1 while in the second half it contains chip N. Thus, section 124 can be said to contain a later-in-time portion of the input signal while section 126 can be said to contain an earlier-in-time portion. In time periods subsequent to t.sub.c, input register 102 would contain chips N+2, N+3, etc.
A reference code is generated by a code generator (not shown) and transmitted via a line 104. This reference code comprises the same pattern of chips used to transmit the input signal. Thus, it is expected that the chips received by the input signal are the same as the chips of the reference code. In this example, chip N is transmitted by the reference code during time t.sub.c.
During the time period t.sub.c, whatever chips are in sections 124 and 126 are multiplied with chip N of the reference code. Specifically, the reference code is multiplied with the chip in section 124 using multiplier 108 and the reference code is also multiplied with the chip in section 126 using multiplier 110. Typically, multi-sampling would occur where several multiplications actually take place during the period t.sub. c. Although 1's and 0's are actually multiplied, it is conceptually easier to envision 0's as -1's. Consequently, since [reference code chip] N×(input register chip) N=1, and [reference code chip]N×(N+1 or N-1) would have a 50--50 chance of equaling a 1 or a -1, then if the input signal is aligned with the reference code the number of 1's resulting from multipliers 108 and 110 should be about the same over a given period of time.
If, however, the input signal is not aligned (i.e., not synchronized) with the reference code, then during time period t.sub.c, chip N will be located in either section 124 or section 126 of input register 102 for a longer period of time than in the other section. For example, if the input signal is running more slowly (i.e., it is "late") as compared with the reference code, then chip N will be located in section 124 for a longer period of time than in section 126. If multi- sampling is used within time t.sub.c (and also when subsequent time periods are looked at) then multiplier 108 will yield more 1's than multiplier 110 during this time period. Conversely, if the input signal is running more quickly (i.e. , it is "early") as compared with the reference code, then multiplier 110 will yield more 1's than multiplier 108.
Another way of looking at this concept is now described with regard to FIG. 1B. Referring to FIG. 1B, the chips within input register 102 are shown aligned with each other and with the reference code during time period t.sub.c. If the input signal is received too quickly with respect to the reference code, the input signal would "shift" to the right. Therefore, chip N of the reference code would be multiplied with chip N via the "early" part of the input signal (in section 126) more frequently than with chip N via the "late" part of the input signal (in section 124). Thus, more 1's would result from multiplier 110 than from multiplier 108. The greater the difference in 1's, the greater the misalignment.
Thus, the values of the multipliers can be used to determine the direction as well as the magnitude of any misalignment between the input signal and the reference code. However, because of the random nature of the products generated by multiplying N×-1) and N× (N+1) as discussed above, some scheme is also typically used to average or "smooth" out the results of multipliers 108 and 110 so that a more accurate assessment of the alignment situation can be determined. Such a scheme is explained with regard again to FIG. 1A.
Referring back to FIG. 1A, an adder 112 adds the values from multiplier 108 and multiplier 110. It is contemplated that the adder 112 yields a positive sign if the results of multiplier 108 are positive and a negative sign if the results of multiplier 110 are positive. In this way, a signed value is generated corresponding to whether the input signal is running earlier or later than the reference code.
An integrator 114 is then used to actually "smooth out" what would otherwise appear as "noisy" signals. Consequently, the integrator typically obtains its output value by sampling the input signal over many chips. The output of the integrator 114 thus indicates an average of both the direction and magnitude of misalignment of the input signal. This output is then typically used to adjust the rate of the reference code to bring it into alignment with the input signal so that synchronization can be maintained.
A problem arises with conventional delay lock loops in post- demodulation environments, however, when data is embedded in the input signal. This is because the data is embedded by inverting the pattern in which the signal is sent. In other words, a non-inverted pattern might be equivalent to a binary 0 while an inverted pattern might be equivalent to a binary 1. The effect of this is that the data affects the ability of the reference code to match the pattern of the input signal. More specifically, many of the chips in the input signal will no longer be the same as the corresponding chips in the reference code. Because of this, synchronization cannot be maintained due to changes in the pattern due to data.
Another problem with conventional delay lock loops is that the electromagnetic waves transmitting the data may be using phase modulation. The results of this is that the input signal may have been inverted during transmission. If that is the case, then any misalignment readings will also be inverted (i.e., a "late" input signal will be interpreted as an "early" one).
Consequently, what is needed is some scheme to detect the direction of a misalignment of an input signal with regard to a reference code regardless of whether data is embedded in the input signal, and regardless whether the input signal has been inverted due to phase modulation.
SUMMARY OF THE INVENTION
The present invention fulfills the needs discussed above by providing a system and method for detecting a pre-determined pattern in the chips of a demodulated input signal by comparing the chips of the input signal with a reference code. The reference code contains a portion of a pre- determined pattern used by the signal transmitting device to indicate that data is forthcoming. If chips in the input signal match the pattern portion in the reference code, then subsequent chips from the input signal (which may or may not be contiguous to the initially detected pattern portion) are examined for the same pattern portion or some mathematically derived pattern related thereto. Detection of some pre- determined number of pattern portion (or mathematically derived) matches indicates that the entire transmitted pattern has been received and that data transmission is forthcoming. By transmitting related pattern portions, the present invention effectively extends the length of a transmitted pattern without substantially increasing the circuitry of the pattern detecting mechanism.
Once the above-mentioned pattern has been received (and initial synchronization has been obtained), data is then transmitted embedded in the input signal. The present invention allows synchronization to be maintained during this phase by determining whether a chip in the input signal has been changed as a result of embedded data and adjusting for any resulting error in indication of misalignment direction. The rate of the reference code is then adjusted accordingly. Thus, the present invention allows synchronization to be maintained with an input signal during transmission of data regardless of whether data is embedded within the input signal.
Other embodiments of the present invention further contemplate that synchronization can also be maintained regardless of whether the input signal has been inverted due to phase modulation.
BRIEF DESCRIPTION OF THE DRAWINGS
Various objects, features, and attendant advantages of the present invention can be more fully appreciated as the same become better understood with reference to the following detailed description of the present invention when considered in connection with the accompanying drawings, in which:
FIG. 1A is a diagram of a conventional delay lock loop.
FIG. 1B is a timing diagram comparing an input signal with a reference code.
FIG. 2 is a high-level block diagram of embodiments contemplated by the present invention for obtaining synchronization.
FIG. 3 is a flow diagram of a method contemplated by embodiments of the present invention for obtaining synchronization.
FIGS. 4A and 4B are block diagrams of embodiments contemplated by the present invention for obtaining synchronization.
FIG. 5 is a block diagram contemplated by embodiments of the present invention for maintaining synchronization.
FIG. 6 is a flow diagram of a method contemplated by embodiments of the present invention for maintaining synchronization.
DETAILED DESCRIPTION OF THE EMBODIMENT
The present invention relates to obtaining and maintaining synchronization with an incoming signal prior to and during the transmission of data. More specifically, the present invention relates to recognizing that a pattern within an incoming demodulated electromagnetic signal has repeated itself (or that some mathematical relationship thereof has been received) a specified number of times, indicating that synchronization has been achieved and that transmitted data is forthcoming. The present invention also more specifically relates to maintaining synchronization with the incoming demodulated signal while data, which is imbedded within the signal, is received.
Embodiments of the present invention for obtaining synchronization are now described with regard to FIG. 2. Referring to FIG. 2, an input signal is received via a line 208. As discussed above in the Background section, the input signal is comprised of "chips" derived from information from a demodulated electromagnetic carrier.
A chip comparator 202 compares the chips of the input signal with a reference code via a line 210. Both the chips of the input signal and of the reference code are being received at some given rate. This reference code is representative of a portion of an overall pre-defined digital pattern being looked for in the input signal. Ultimately, a match of this overall pattern indicates that synchronization has been achieved and that data is forthcoming.
Embodiments of the present invention contemplate that the reference code is generated by a code generator (not shown) which is contemplated to generate (or already contain) a pattern which is a portion of the overall pattern being looked for in the input signal. Some embodiments of the present invention contemplate that the overall pattern is a pseudo- random code.
If the chips received via the input signal in line 208 match the pattern (i.e., the pattern portion) received via line 210, then the chip comparator 202 sends a pattern signal via line 214 to a pattern signal detector 206. Embodiments of the present invention contemplate that the pattern signal detector 206 can be set to look for (i.e., count) any number of signals from the chip comparator 202 before the pattern signal detector 206 generates a synchronization signal via a line 212 indicating that synchronization has been detected. In other words, these embodiments contemplate that the pattern must be detected in the input signal a pre- determined number of times before the pattern signal detector 206 will send out a signal indicating that the overall pattern has been detected and that synchronization has been achieved. Extending the effective length of the overall pattern in this manner allows false detections as well as the amount of circuitry to be minimized.
The present invention also contemplates embodiments where after the chip comparator 202 indicates that a first pattern portion in the input signal was found, the subsequent pattern portion looked for is some mathematical derivative of the first pattern portion. For example, after detecting the first pattern portion, the chip comparator 202 may then be set to detect the inverse of the first pattern or the exclusive OR of the first pattern portion. Further, embodiments of the present invention contemplate that the match between the pattern of the input signal and the reference code may be within some pre-defined tolerance for there to be an indication from the chip comparator 202 that a pattern match has been detected. Also, embodiments of the present invention contemplate that the repetitive detections made by the pattern signal detector 206 must occur at predetermined intervals.
Thus, the present invention can detect a long overall pattern in an input signal by actually looking for two or more pattern portions. These pattern portions can be repetitions of the initial pattern portion or some mathematical derivative thereof.
Embodiments of a method of operation of the present invention concerning obtaining synchronization are now described with regard to FIG. 3. Referring to FIG. 3, the first step is that an input signal is received, as indicated by a block 302. As indicated above, this input signal can comprise one or more binary chips. The input signal is then compared with the reference code, as indicated by block 304.
If a pattern in the input signal and the reference code (or some mathematical relationship derived therefrom) are not the same (or are not substantially the same) then the input signal will merely continue to be received as indicated by a decision block 306 and block 302. If, however, the input signal and the reference code are the same (or substantially the same or have some mathematical relationship) then an indication that the pattern has been detected will be generated as indicated by a block 308. Then, if the number of detections of the pattern (or the mathematical relationship derived therefrom) are sufficient, the entire overall pattern is found and synchronization is achieved as indicated by a decision block 310 and a block 312. If, however, the number of detections is not yet sufficient, then the input signal will continue to be received in search of a pattern to match the reference code, as indicated by decision block 310 and block 302.
Although embodiments of the present invention contemplate that the input signal is a demodulated signal derived from the digitization of a wireless electromagnetic wave form, it should be understood that other embodiments of the present invention contemplate operation in any other environment where the input signal is derived from some source where noise and interference exist.
Some detailed embodiments envisioned by the present invention are now described with regard to FIGS. 4A and 4B. In these embodiments, it is contemplated that the initial pattern portion looked for in the input signal is 31 chips in length and that once this initial 31-chip pattern is detected, at least one pattern portion expected thereafter is the inverse of this initial pattern. Thus, the effective length of the overall pattern is at least 62 chips in length.
Referring now to FIG. 4A, the input signal is read into an input register 404 (which is 31 chips in length) via a line 402. Embodiments of the present invention contemplate that the input register 404 is a shift register comprising flip-flops, wherein the chips from line 402 are shifted in one chip at a time. That is, the present invention performs a comparison with the reference code (as described below) each time a new chip is shifted into the input register 404.
A reference code generator (not shown) is envisioned to contain (or generate) at least one of the patterns which is to be searched for in the input signal. A coefficient register 410 receives the reference code (comprising chips) via a line 408.
Each time a chip is shifted into input register 404, each chip residing in the input register 404 is multiplied with its corresponding chip in coefficient register 410 using a multiplier array 406 comprising a plurality of multipliers. Specifically, bit 0 of the input register 404 is multiplied with bit 0 of the coefficient register 410, etc. Some embodiments of the present invention contemplate that each multiplier in multiplier array 406 is a modulo 2 adder.
Once each of the chips is multiplied together as described above, the results of all these multiplications are summed by a summer 412. Embodiments of the present invention contemplate that this summer comprises full adders and half-adders.
The output of the summer 412 appears on line 413. The greater the number of matches between the chips of input register 404 and coefficient register 410, the greater the number that will appear at line 413. For example, if all of the chips matched, then the output on line 413 would be 31. If none of them matched, the output would be 0.
A zero adjust circuit 414 converts the value on line 413 from a unipolar value to a bipolar (i.e., "polarized") value. More specifically, the length of the input register 404 (which in this case is 31) is divided in half so that any value received from line 413 above the half- way mark (in this case, 15) is converted to a positive number (subtracted by 15), and values below the half-way mark are converted to negative numbers. For example, a value of 15 received via line 413 is converted to 0, a value of 17 is converted to +2, and a value of 12 is converted to -3. An absolute value converter 416 (designated |A. vertline.) then converts the polarized value obtained from the zero adjust circuit 414 into an absolute value (output at line 418) and the sign of the polarized value (output at line 420).
A description of the embodiments described above is continued with regard to FIG. 4B. Referring now to FIG. 4B, the absolute value output from absolute value converter 416 branches from line 418 to form lines 418A and 418B. Line 418A is received by a 3 sample input register 421. As with the input register 404, values are shifted into 3 sample input register 421 one chip at a time. Thus, each time a new chip is shifted into input register 404 and the above-mentioned operations take place to produce a new absolute value along line 418A, the new absolute value is shifted into 3 sample input register 421. The result is that 3 sample input register 421 contains not only the absolute value resulting from the latest above-mentioned operations, but it also contains the preceding two absolute values obtained from the preceding two operations. In addition, the absolute value from the current summation is also received by a comparator 430 via line 418B.
In seeking a pattern match, the purpose of the 3 sample input register 421 and associated circuitry is to recognize when the middle value in the 3 sample input register 421 is significantly larger than the other two. Specifically, when a match (or near match as explained below) occurs between input register 404 and coefficient register 410, the absolute value received via line 418 should be (or be close to) 15. On average, when the pattern in coefficient register 410, as a whole, does not match, then on average about half of the individual chips will match. In that situation, the absolute value transmitted at line 418 would be (or be close to) 0. Thus, by looking for such a "peak" in the middle value (i.e., at "S1") there is less possibility of a false synchronization detection.
Circuitry associated with some embodiments of the 3 sample input register 421 of the present invention is now described. To begin with, the current value shifted into the 3 sample input register 421 ("S2") is not sampled from this register 421. That is, only values "S0" and "S1" are sampled. Instead, the current value is used as an input to comparator 430 along line 418B. The other two values in 3 sample input register 421 (that is, S1 and S0) are received as inputs to comparators 426, 428 and 430. The results of these comparators are then input into an AND gate 432.
In essence, comparators 426, 428 and 430 along with AND gate 432 determine whether the second absolute value (Si) in 3 sample input register 421 has created a "peak" as compared with the first absolute value (SO) and the current absolute value to indicate that the pattern being looked for has, indeed, been detected. In some embodiments of the present invention, the synchronous threshold line 424 (which is an input of comparator 430) determines the magnitude that the "peak" must be for the AND gate 432 to output a signal indicating that the pattern has been found.
Once it has been determined that a peak has been detected, a peak detect signal is received along line 440 from the AND gate 432 by a code sync detector 434. The sign of the value transmitted from the zero adjust circuit 414 is also received via a line 420. The significance of this is that if the chips in input register 404 and coefficient register 410 match, then the sign received via line 420 is positive.
Thus, in embodiments contemplated in the present invention where a matching of the pattern is sought first and its inverse is then looked for, the sign received from line 420 will be positive upon detection of a match and a signal from line 440 will also be received. When this happens, a code period timer 436 is activated and counts down some pre- determined time period. At the end of this time period, the code sync detector 434 looks for a peak detection signal from line 440 and a negative signal from line 420. Embodiments of the present invention contemplate that this time period is equal to the time required for the input register 404 to shift in an entirely new pattern's-worth of chips (in the embodiments of FIG. 4B, this is the time required for the next 31 chips to be received by input register 404).
As indicated above, these embodiments contemplate that these next 31 chips should be the inverse of the initially matched pattern. If this is the case, none of the chips in input register 404 will match the pattern in coefficient register 404, and the result at line 413 will be approximately 0. Assuming the value is in fact 0, the zero adjust circuit 414 will adjust this value to -15. The absolute value converter 416 again outputs a +15 along line 418, but this time a negative indication is output along line 420. Thus, code sync detector 434 receives a peak detect signal from line 440 and a negative signal from line 420. Since this is what the code sync detector 434 is looking for, the code sync detector 434 outputs a synchronization detected signal along line 438 indicating that the overall pattern has been found.
If the code sync detector 434 does not receive the expected inputs in the designated amount of time, then it assumes that the first pattern match was just a false alarm and the present invention begins to look for the initial pattern again.
As indicated above, it should be understood that embodiments of the present invention contemplate that any number of mathematically related patterns could be looked for (one after the other or in some other fashion) and that the present invention is not limited to looking for the initial pattern and its inverse. Also, the present invention contemplates embodiments where the 3 sample input register 421 could be of other lengths and that the number of chips received via the input signal could have been counted rather than using code period timer 436. In addition, embodiments of the present invention contemplate that other circuits, circuit designs and configurations to perform the above-noted functions could also be used and are contemplated by various embodiments of the present invention.
Some embodiments of the present invention contemplate use in environments where the inverse of the initial pattern (as well as any mathematically related patterns thereafter expected) may be inverted due to phase modulation. In these embodiments, the present invention nonetheless recognizes the initial pattern from the input signal as a valid indication that data is forthcoming, and just assumes that all subsequently received patterns or mathematically related patters) will also be inverted. Also, some embodiments contemplate that above- described scheme for obtaining synchronization can be used in pre- demodulation environments as well.
Once synchronization is achieved, embodiments of the present invention contemplate use in environments where data is subsequently received via the input signal. It is envisioned that this data is received in the form of a pattern of chips as discussed in the Background section above. That is, receipt of a specified pattern might indicate a binary 0 and receipt of some mathematical representation thereof (e.g., inversion of the pattern) might represent a binary 1.
The way that embodiments of the present invention contemplate that synchronization is maintained using a pattern within the input signal with data embedded within the input signal is now described with regard to FIG. 5. Referring now to FIG. 5, the output of adder 512 and preceding circuitry works in the same way as described in the Background section above. Specifically, a section 524 of an input register 502 (containing a chip as shown in section 124 of FIG. 1A) is multiplied by the reference code via multiplier 508, while a section 526 of input register 502 (containing a chip as shown in section 126 of FIG. 1A) is multiplied by multiplier 510. If the signal is shifted "late" then the results of adder 512 will be positive. If, however, signal is shifted "early" then the results of adder 512 will be negative. Thus, adder 512 generates a misalignment signal. Of course, the present invention contemplates modified designs to achieve the same basic function. In any event, as indicated above, these components alone are insufficient to maintain synchronization when data is embedded in the input signal.
To resolve the problem of maintaining synchronization during receipt of embedded data, embodiments of the present invention envision the existence of another section 528 of input register 502. This section 528 is contemplated to contain chip N itself during the time period t. sub.c as mentioned in the Background section. That is, the chip in section 528 and the chip of the reference code received via a line 504 at t.sub.c should be the same.
At approximately the middle of period t.sub.c, some function is performed on the chip of section 528 of the input register 502 and the chip of the reference code received via line 504. The precise operation depends upon how the pattern of the input signal is affected when data is embedded. For example, if the pattern by itself represents a 0 and an XOR of the pattern represents a 1 (or vise versa), then the operation would be another XOR. Some embodiments of the present invention contemplate that an inversion of the pattern represents a 1. These latter embodiments are assumed during the discussion below.
For each chip appearing at section 528 during period t.sub.c, a multiplier 520 multiplies the chip in section 528 with the chip received via line 504. If the two chips are identical, the result is a 1. If the two chips are different, then the result (which is 0) is treated as a -1. In essence, the two chips are compared for identity and the result is effectively a direction correction signal.
The result of the above-mentioned multiplication is then multiplied by the output of adder 512 using a multiplier 516. This multiplication causes the result of the adder 512 to reverse polarity. This consequently has the effect of canceling the effect that the embedded data has on determining the direction of the misalignment between the input signal and the reference code. Thus, the misalignment signal becomes a corrected misalignment signal.
Environments contemplated for use with the present invention contemplate that the data received via input signal is extracted for use using some device that is separate from the embodiments shown in FIG. 5.
Some embodiments of the present invention also contemplate that a polarity latch 522 is used to compensate for inverted results of adder 512 due to inversion of the pattern caused by phase modulation of the electromagnetic carrier. These embodiments contemplate that the polarity latch 522 receives some indication (such as, for example, line 420 of the embodiments of FIG. 4A for obtaining synchronization discussed above) of whether the entire input signal has been inverted. If it has, then the polarity latch generates a -1 which is then multiplied by the output of adder 512 using a multiplier 518.
The output of adder 512 (as possibly modified by multipliers 516 and 518) are then received by integrator 514 (which might be, for example, an "integrate and dump" type low pass filter) which "smooths" the results as discussed above so that the rate of the reference code can be adjusted to bring the reference code into alignment (i.e., maintain synchronization) with the input signal. Specifically, in the embodiments shown in FIG. 5, a positive error output from integrator 514 indicates that the rate of the reference code should be decreased and a negative error output indicates that the rate should be increased.
Operation of various embodiments of the present invention are now described with regard to FIG. 6. Referring now to FIG. 6, the first step is that an input signal is received, as indicated by a block 602. The next step is that the contents of the early (526), late (524) and reference (528) sections of input register 502 are multiplied with a corresponding portion of the reference code. This is indicated by a block 604.
The next step is to sum the results of the two multiplications involving sections 526 and 524. This is indicated by a block 606. This sum is then multiplied with the result of the multiplication involving section 528 and the reference code, as indicated by a block 608. The next step is to integrate to determine the magnitude and direction of any misalignment. This is indicated by a block 610.
It should be understood that in operating on various chips and other values located within the various registers mentioned above, embodiments of the present invention contemplate that over-sampling techniques are used. Also, various embodiments contemplate that storage other than registers (for example, RAM or some magnetic/optical media) could also be used in place of the registers or in place of any mechanism for storing values.
Some embodiments of the present invention contemplate that a "sample and hold" circuit is used to take a kind of "snapshot" of the output of multiplier 520. More specifically, this "snapshot" is taken at time 1/2 t. sub.c. This allows for a more accurate reading of the chip in section 528 at time t.sub.c.
Some embodiments of the present invention also contemplate that the reference code received via line 504 (as well as the corresponding pattern used to transmit data received via the input signal) is different (and longer) than the reference code received via line 210 or 408. Further, some embodiments envision use in environments where transmission of data is somewhat delayed after transmission of the pattern used to achieve initial synchronization and that several iterations of the pattern otherwise used to transmit data are sent without data embedded in them. This is to allow embodiments of the mechanism shown to FIG. 5 to adjust (i.e., lock on) to the receipt of the input signal. Some of these embodiments envision that 172 chips is sufficient to make this adjustment.
Although the embodiments discussed above concern the use of digital technology, it should be understood that embodiments of the present invention contemplate analogue equivalents where a demodulated (but undigitized) input signal is processed as described above.
It should be emphasized that the various components of embodiments of the present invention can be implemented in hardware, software or a combination thereof. In such embodiments, the various components and steps would be implemented in hardware and/or software to perform the functions of the present invention. Any presently available or future developed computer software language and/or hardware components can be employed in such embodiments of the present invention.
It should also be understood that the present invention is not limited to the embodiments indicated above, and that the examples presented above are merely for the purposes of illustration. The scope of the present invention should therefore be interpreted by the following claims as defined by the forgoing figures and text. | |
The Galmont's Statement on Coronavirus (Covid-19)
Dear Guest,
We are delighted to have been awarded the Covid 19 Safety Charter by Fáilte Ireland. This is to confirm that we have completed Fáilte Ireland’s Infection Prevention Control for Tourism Businesses course.
We are pleased to let you know that we have introduced new physical distancing guidelines and advanced sanitation measures to provide a safe and comfortable guest experience for you. We want to reassure you that we are closely following the guidance and procedures of the World Health Organisation and the Irish Health Services Executive (HSE) regarding the Coronavirus, to support the health, safety and wellbeing of our Guests, our Staff, and our Community.
We take standards for hygiene and cleanliness very seriously and are proactively implementing additional hygiene procedures and precautions to ensure that the highest possible standards are upheld throughout every area and service which the hotel provides, for you our guests, as well as our own team.
This includes hand sanitisers in all areas, deep cleaning of all space during and after departure, and the reduction of seating in social areas for all refreshment and meal periods.
We are supporting our hotel team, and their own health, safety and knowledge, as these are essential to an effective cleaning program. Here are some ways in which we are supporting them:
- Face masks: We ask that all of our guests wear a face covering when entering and moving around the hotel.
- Hand hygiene: Proper and frequent handwashing is vital to help combat the spread of viruses. In our daily hotel and departmental meetings, our teams are reminded that cleanliness starts with this simple act. It’s important for their health and that of our guests.
- Ongoing training: In addition to training on housekeeping and hygiene protocols, hotel associates have also completed enhanced COVID-19 awareness training.
- Real time information: The Hotel’s corporate and regional teams are on standby 24/7 to support the Hotel and coordinate with local and regional authorities.
- Cleaning products and protocols: Our hotel uses cleaning products and protocols which are effective against viruses.
- Guestrooms: The Hotel uses cleaning and disinfecting protocols to clean rooms after guests depart and before the next guest arrives, with particular attention paid to high touch items. The rooms are cleaned using colour coded microfibre cloths, five different cloths per room for each area, these will be changed between each room, the sanitiser chemical is “Suma Bac D10” manufactured and supplied by Johnson Diversey and is certified to kill Corona Virus. The accommodation assistants will be wearing facemasks and gloves. Once the guestroom is cleaned, the high touch areas will be sanitised again with D10.
- Public spaces: The hotel has increased the frequency of cleaning and disinfecting in public spaces, with a focus on the counter at the front desk, elevators and elevator buttons, door handles, public restrooms and even room keys.
- Reception: Social distance markers have been put in place in our Reception and Lobby areas, to control check-in queues and general crowding. Screens have been put in place at the Reception Desks. All credit card machines are sanitised after use.
- Restaurant and Bar: Tables are sanitised after each guest visit. All seating arrangements have taken into consideration the recommended social distancing. We have introduced single use menus. All credit card machines are sanitised after use. All dining times must be booked in advance of your stay, including breakfast. We wish to advise that in line with Government Covid-19 guidelines & restrictions we cannot serve any drinks past 11pm.
- Energize Fitness & Leisure: One hour sessions in the gym and swimming pool must be booked in advance. Customer Sanitisation Stations are located inside the entrance of Energize Fitness & Leisure, entrances to changing rooms and throughout the gym, enabling members and guests to regularly sanitise their hands. Anti-bacterial sprays are located around the gym for guests and members to sanitise equipment before and after use. Energize Fitness & Leisure will close for 30 minutes between each hourly session, to allow team members to sanitise the gym, gym equipment and changing rooms using a electrostatic misting sprayer. This dispenses a food grade sanitiser provided by Johnson Diversey called Suma Bac D10. The steam room will remain closed until further notice.
- Spirit One Spa: Treatments are limited and must be booked in advance. Thermal suite will remain closed until further notice.
- Carpark: Our onsite carpark allows convenient parking with direct access to the hotel lobby.
We will continue to keep you up to date and we look forward to welcoming you to The Galmont Hotel soon. | https://www.thegalmont.com/en/coronavirus-statement-the-galmont/ |
Alison Eastwood is an actress, film director and producer but most importantly an animal advocate. She grew up in Carmel, California where she spent her childhood riding horses, which instilled a deep love and respect for animals and nature. She is an active member of several animal welfare organizations and donates her time fundraising and volunteering to help save animals. Alison has been actively involved in trying to ban the the killing of wild horses in Nevada and is also the spokesperson for Marine Animal Rescue of Los Angeles. She is spearheading their fundraising efforts to build a new marine mammal care center. She has also been involved in fundraising for Chimp Haven, Farm Sanctuary, and Born Free.
Alison is also the Founder of the Eastwood Ranch Foundation. The Eastwood Ranch Foundation provides support to animal welfare groups, non-kill shelters, rescues and sanctuaries around the U.S. by providing funding, support and logistical assistance to help better manage their organizations. They rescue domestic animals from high kill shelters in southern California while supporting other vetted local rescues and providing care until a forever home is found. Besides local animal rescue, ERF works on behalf of any and all animal welfare causes by helping raise awareness, educating, campaigning/petitioning for Animals throughout the United States and beyond. | https://petphilanthropycircle.com/alisoneastwood.html |
BONN, Germany (Landscape News) — Along with rising sea levels and disrupted weather patterns, climate change could also have a potentially harmful impact on migratory birds. That finding comes from a new study carried out by ornithologists at Cornell University, recently published in Ecology Letters.
The study used computer models of climate change predictions in the Western Hemisphere and compared them to weekly population estimates for 77 species of migratory birds that fly between North America and Central and South America. The scientists based those estimates on 13 years of observations from the eBird citizen-science database, an online program run by the Cornell Lab of Ornithology.
They also examined three climate variables from the computer modeling: minimum and maximum surface temperatures at two meters above the ground, and surface precipitation rates.
Previous studies have looked at novelty – deviation from historical climate norms – year by year, said Frank La Sorte, a research ecologist at the Cornell Lab of Ornithology and lead author of the paper.
“This is the first study that looks at novelty seasonally,” he said. “We are seeing that novelty actually shows some interesting dynamics when you look at it from one season to the next. And the dynamics carry interesting implications for birds and other species that would not be obvious just looking at annual summaries.”
According to the paper, there was greater deviation from novelty in the birds’ tropical non-breeding grounds than in their temperate breeding grounds in the North.
However, the paper states: “contrary to expectations, highly novel climates also occurred within temperate regions during the transition from breeding to autumn migration.”
The study saw “a pronounced peak occurring when juveniles are leaving the nest and preparing to embark on their first migratory journey, which may adversely affect their chances of survival.”
“This is a critical period, when juveniles are leaving the nest and both adults and juveniles are preparing for the long migration journey south,” said La Sorte.
Change in the autumn climate could threaten plants as well as insect communities. “So if insect resources are changed in terms of abundance, or species composition that could affect their ability to fulfill the basic requirements during that period, which might reduce survival,” he explained.
The implications are especially dire for eastern bird species. They need to both bulk up on insects to prepare for the long flight, and take advantage of the autumn weather conditions.
“They have to travel further, and many of them will cross the Gulf of Mexico or the Atlantic Ocean,” said La Sorte, “So they require either mild or supportive winds to make that journey efficiently. Birds in the West winter in Mexico. They don’t have to travel as far and they don’t have to cross any large bodies of water.”
If eastern bird species need more time to build up their fat layers, they will lose out on those generally favorable atmospheric conditions for flight. “The longer the wait, the worse the weather is going to get,” he said.
At greatest risk are juveniles preparing for their first migratory journey in the autumn, he added. “If they are adversely affected, that could affect the overall population size. In the long term, that could be detrimental.”
Having pinpointed locations and times when migratory bird populations face the greatest challenges from climate change, the scientists hope that information from their study will help identify when and where conservation efforts will have the greatest benefit.
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The portraits of artist Ryan Hewett invoke an uncanny sense of familiarity and mystery, as each painting leaves you searching the faces of his subjects for their true identity. The artist uses thick brush strokes and heavy paint to build up compelling palettes across his subject’s facial features. Dense globs of pinks and reds are built up, forming an impression of each person’s identity. In his series Order, Hewett’s impasto technique provides a stark contrast to the smooth, soft texture that is often present in his backgrounds as well as the bodies of his subjects. Hewett’s incredible style brings to mind the fleshy, expressive paint strokes present in the work of Frances Bacon.
There is a psychological element to Hewett’s portraits that is as beautiful as it is distinctive. Hewett’s website explains, “His portraits are not life-like depictions, but rather abstracted representations of his subjects.” Perhaps this abstraction reflects the subject’s inner psyche rather than their actual identity. Although the viewer cannot see the details of the individual person’s identity, something else can be sensed instead. An element of secrecy and intrigue is held in each thick brush strokes, hiding what is just beneath the surface.
The South African artist has shown his captivating portraits all over the world in cities like Berlin, Miami, Cape Town, London and New York City. | https://www.createmagazine.com/blog/2017-8-11-ryan-hewett |
Description
The fourth entry of the Atelier Iris series, this time around the player takes control of a student named Vayne Aurelius at the Al Revis Academy for Alchemy, where players complete tasks and quests set before them by a professor of the university. If a player fails to complete these tasks they will have to stay behind and also the story will change depending on who the player chooses to work with during these quests.
The game still features RPG-style elements, but now with different features. Instead of "leveling up" by defeating enemies where the player can choose how there character will develop using a "crafting" system, letting them choose which attributes they want to modify. Players can also "craft" items and weapons using various objects scattered throughout the world.
Players will no longer have random engagements, something more common in many RPGs, this time around the player can choose who or what they want to fight with the ability to see your opponents symbols on the board, choosing if they want to engage them.
The PSP version features multiplayer battles, more items that can be synthesized, and an option to reduce loading times.
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You chose your path in the legal field, then you definitely need to read these 5 tips.
Article By: Omar Naguib
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Understand the Changing Profession…and Adapt.
This profession is not like what it used to be. And the legal field continues to evolve, which is unnerving today’s lawyers. Instead of sticking your head in the sand and pretending that it’s yesteryear, be practical in understanding the direction of this change so you can better prepare yourself to remain relevant, secure and marketable. How this can be done? For example, one can: read the news, analyze the job boards, join local executive/legal groups and join meetings on current events; and stay linked with your network. Always try to encounter the new changes and try to benefit from it as much as possible
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Learn How To Communicate Effectively.
This is about the words, tone and messages you use when you are speaking with or writing to another person – at any-time, for any-thing. You should learn how to persuade the person/s you are talking to by your point of views
Words and tone are powerful. And the skill a person possesses when using them can be the difference, receiving a higher offer or getting it pulled, keeping your client or losing it. Effective communication is also about being concise by not to talk in irrelevant issues, always try to consider only relevant issues …and relevant. Also an example is when a lawyer knows all facts about case and knows its solution, although he doesn’t possess the skill of effective communication, he might lose his case. So it is very important to know how to communicate in the most effective way.
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Perform Self-Assessment
Before opting of the legal field you would like to engage in, it is critical to put self-assessment for yourself, you should figure out your points of strengths and weaknesses which will aid you to know which legal field is optimum for your knowledge and skills and to know which field will you be able to be productive and efficient in. This self-assessment should be done regularly to ensure that you are on the right track.
During assessment, when considering weaknesses, try immediately to avoid it so for example if you are somehow week in legal research, take legal research courses to have this crucial skill. One thing needs to be taken into consideration is that you will never be 100% perfect, you should always improve yourself and never stop.
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Shadow a Legal Professional
Want to overcome practical life challenges? Shadow a legal professional. To clarify, job shadowing is the gate for entering practical life , by shadowing , you can see what tasks you could be assigned to, how could the workplace atmosphere be , what you should avoid in work to succeed and what you should do. Moreover, it’s a great chance to ask the legal professional about the job to know much more details that might change your choice such as amount of work and people you will deal with on daily basis.Furthermore, Shadowing enables you to live a typical work day, and this prepares you when entering practical life, to be professional and able to deal with numerous situations.
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Become tech-savvy
Technology updates and revolutionizes the legal industry. It would be great skill for a legal person to be able to use technology in efficient way and be able to adapt to the new updates so that you can use these updates to improve your technological skills which shall consequently help you to find what you need. A clear example is websites that you can just enter the title you need to find cases about and cases will appear to you without doing the effort and consuming a myriad of time, so when client seek your legal opinion and find that you are up to date with technology, this will be appreciated, unlike when you lack technology, you will consume a lot of time, money and effort to reach what you aim to find. | https://thelawmag.com/2019/07/31/5-tips-to-help-law-students-in-their-career-path/ |
Earth’s flora and fauna have over the last 540 million years seen mass extinction on five occasions, with an elevated rate of extinction and loss of at least 75 % of the species in less than two million years. The Ordovician, Devonian, Permian, Triassic and Jurassic geological periods all ended with a sharp decline in all orders of species. The mass extinctions were caused by a combination of climate change, changes in the composition of the atmosphere, greatly altered habitats, and in the seas and oceans both acidification and lack of oxygen. The fifth mass extinction, which probably took place over only a few years, saw the disappearance of all large dinosaurs. Many researchers claim that we are now standing on the brink of the sixth mass extinction, this time caused by man. Is it so? To answer this question we need to look at the current speed and scale of the extinction.
How fast is the rate of extinction?
The normal rate of extinction has been estimated to be between 0.1 and 2 species per million per year. The number of documented extinctions in history (since 1500) is only about 1,000 species, which does not sound much considering that just over 1.7 million species have been described, but it is still sufficient to be regarded as an abnormal increase in the rate of extinction. Among the animals we know best, the vertebrates, 200 species have become extinct over the last 100 years. With a normal rate of extinction this would have taken up to 10,000 years, not 100. The rate of extinction of vertebrates is already 100 times higher than the normal rate. We know less about other organisms but based among other things on assessments of the number of natural habitats that have disappeared, the current rate of extinction of the Earth’s species is estimated to be between 10 and 1000 times higher than is normal.
An assessment method uses the ratio between number of species and area together with data on known biotope losses to predict the loss of species. 12-24 % of the Amazonian forests are expected to disappear between 2008 and 2050, which would result in the loss of 5-9 % of the plant species. Other methods are based on taking inventory before and after biotope changes and extrapolating to a global scale. All methods have their weaknesses and sources of error, not least because we know about only a fraction of the species that exist. The pattern, however, is clear: some orders of animals and plants now have a much higher rate of extinction than normal. The rate of extinction among birds is 100 times higher than is natural and for amphibians 211 times.
Over 25,000 species are listed as threatened around the world and many of them will die out within the next few centuries, which will further increase the rate of extinction. By about 2100 the rate of extinction among birds will be an estimated 1,500 times higher and among amphibians more than 25,000 times higher. Many of the endangered species are already considered to be unable to survive because too little of their natural habitats remains and they therefore constitute a global extinction debt. Theoretical species-area models applied on losses of forests in South-East Asia indicate that 79% of South-East Asia’s vertebrates will be facing extinction by 2100. If most of the globally threatened species die out within a century the rate of extinction will be at least 1,000 times higher than is natural, which is on a par with earlier mass extinctions, or far above depending on how long it took.
How widespread is the extinction?
The extent of the extinction is also approaching the magnitude of previous extinctions. In many groups of animal and plant species over 25% of the species are threatened. For those groups that are most severely affected by human activities, the proportion of threatened species is even higher: amphibians 42 %, crocodiles 47 %, cycads 63 %, and sturgeon 85 %.
A global loss of species is bad enough but even worse from the point of view of man’s possibilities to take advantage of the ecosystem services the species offer, is the ongoing loss of species locally and regionally, and the decreasing population size and distribution of many species. A large part of the species stock, in particular the large herbivores and predators, can be considered to be functionally extinct. The large-scale pattern is that endemic and specialised species are being annihilated and replaced by global colonists, i.e. the species that are favoured by human activities (including pests and pathogenic organisms). The result is that local sets of species are becoming increasingly similar to each other. The local number of species can thus be maintained, at the same time as the Earth loses diversity among species.
The trend for most threats to species is increasing. There are now 6.7 billion people and this figure will have risen to 9.3 billion already by 2050. All these people need space to live in and resources such as food and energy. This takes living space from animal and plant species. Mankind today uses 25 % of the planet’s primary production, in some regions up to 80 %. Climate change is also expected to become a more serious threat factor over the next 100 years. The “perfect storm” in the form of climate change and substantially altered habitats, including acidification and lack of oxygen in the seas and oceans that characterised the five previous mass extinction periods is already upon us.
The sixth mass extinction is coming closer
As stated earlier we are now standing on the brink of the sixth mass extinction, this time a mass extermination, caused by man. The rate of extinction is already sufficiently high for us to be able to say this, and we are approaching a magnitude of 75 % loss. This mass extinction began already at the end of the ice age, when man contributed to the extinction of the mammoths. If all threatened species of vertebrate die out in the near future and the rate of extinction then reached continues to prevail, 75 % of vertebrates will be gone within 240-540 years. There are several assumptions and rough estimates behind this conclusion, but it is not unreasonable.
The forecasts for future development depend for the most part on how people choose to manage the Earth’s resources. With radical changes, and if they are realised very soon, we can reverse the trend in global extinctions. Scenarios for future developments, however, show a complex relationship between species conservation and climate measures. The scenarios that have the lowest temperature increase are not the best from a biodiversity perspective because large areas would need to be used to produce bio-fuels to replace fossil fuels. The worst scenario, however, is “business as usual”: more severe climate changes and increased land use to feed a growing population.
Following the last five mass extinctions, the diversity of species has recovered but it has taken millions of years. The mass extinction at the end of the Cretaceous period was among the most severe in the Earth’s history, but recovery was in the context very fast; within 300,000 years local species’ diversity had been restored counted as number of species, even though the fauna and flora at that time were quite different. What will come after the sixth mass extinction? Life will probably persist, but will man? | http://www.biodiverse.se/articles/the-sixth-mass-extinction/ |
A letter from your Higher Self Exercise
You can imagine your Higher Self as a friend who is unconditionally wise, loving and compassionate. Imagine that this friend can see all your strengths and weaknesses, including wonderful qualities you don’t always recognize in yourself. This friend understands the limits of human nature, and is kind, accepting, and forgiving.
Now write a letter of encouragement to yourself from the perspective of this imaginary friend (your Higher Self). Remember that your Higher Self can see all the wonderful qualities you have inside of you and knows you have unlimited potential to life your highest joy in life.
What would this friend say to you from the perspective of unlimited compassion and unwavering faith? And if you think this friend would suggest possible changes you should make, how might these suggestions embody feelings of trust, care, encouragement, and support?
Once you’re finished, put this letter in a sealed envelope with a stamp. Ask a friend or family member to write your name and address on the envelope and take good care of it. Ask them to send you the envelope in about 1 or 2 months from now.
When you receive this letter by mail, take your time to read it and let the words sink in. Feel the compassion as it pours into you, soothing and comforting you. Love, connection and acceptance are your birthright. To claim them you need only look within yourself with the help from the perspective of your Higher Self. | https://www.sandrarolus.com/a-letter-from-your-higher-self-exercise/ |
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Displaying 1 - 10 of 45 results.
Senior Research Staff
Maurice Obstfeld
Nonresident Senior Fellow
Senior Research Staff
Edwin M. Truman
Nonresident Senior Fellow
Speeches & Papers
The Decline of the Dollar
C. Fred Bergsten
(PIIE)
January 29, 1999
Working Paper
Postponing Global Adjustment: An Analysis of the Pending Adjustment of Global Imbalances
Edwin M. Truman
(PIIE)
July 2005
Op-Eds
The Fund Appears to Be Sleeping at the Wheel
Morris Goldstein
(PIIE) and
Michael Mussa
(PIIE)
October 3, 2005
Speeches & Papers
Renminbi Controversies
Morris Goldstein
(PIIE)
November 3, 2005
Op-Eds
A New Way to Deal with the Renminbi
Morris Goldstein
(PIIE) and
Nicholas R. Lardy
(PIIE)
January 20, 2006
Testimonies
The US Trade Deficit and China
C. Fred Bergsten
(PIIE)
March 29, 2006
Op-Eds
Exchange Rates, Fair Play, and the "Grand Bargain" | https://www.piie.com/search?sort=field_date&order=asc&%3Bf%5B1%5D=field_contributor%3A1838&%3Bamp%3Bf%5B1%5D=field_related_topics_and_regions%3A977&%3Bamp%3Bf%5B2%5D=field_contributor%3A2345&f%5B0%5D=field_related_topics_and_regions%3A1119&f%5B1%5D=field_related_topics_and_regions%3A260 |
Questions and Answers on Per and Polyfluoroalkyl Substances (PFAS) in Food
Is the food supply safe?
The U.S. food supply is among the safest in the world. The FDA is working to understand the occurrence of PFAS in the general food supply by testing for certain PFAS chemicals, including perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS). To date, we have found that most foods not grown or produced in specific geographic areas contaminated with PFAS do not have detectable levels of PFAS.
For good nutrition and for food safety, the FDA recommends that consumers eat a varied, well-balanced diet. See more information on the Federal Dietary Guidelines website.
How is the FDA addressing PFAS in foods?
To better understand the exposure to PFAS from foods for people in the United States, we are focused on testing foods from the general food supply. We are also providing technical consultation to states, when requested, to help determine if there is a potential health concern for foods that are grown or produced in specific geographic areas contaminated with PFAS. In addition, we are reviewing the current scientific evidence on specific types of PFAS (referred to as “short-chain”) authorized for limited use in certain food contact applications.
As we advance our understanding of the occurrence of PFAS in foods and complete our review of research on short-chain PFAS, the FDA will determine if additional actions are needed to protect public health.
Should I stop eating particular foods to reduce my PFAS exposure?
There is no scientific evidence that supports avoiding particular foods because of concerns regarding PFAS contamination. Foods that are associated with areas of environmental contamination may or may not pose a risk. Research has shown that PFAS contamination in the environment where food is grown or produced does not necessarily mean the food will contain detectable levels of PFAS. This is because the amount of PFAS taken up by foods depends on many factors, including the specific type of PFAS and type of food.
How does the FDA determine if a food from a contaminated area is safe?
When states identify foods that are grown or produced in a specific geographic area of contamination, they can contact the FDA to request technical assistance. This includes analyzing samples but also assessing the safety of levels found, if any. We are working with local and state partners, as well as other federal agencies, to determine the actions needed to address any food safety risks and, if necessary, prevent the food from entering the marketplace if determined to be a health concern.
What levels of dietary PFAS exposure cause adverse health effects in humans?
Most of the research on PFAS and health effects is based on two types of PFAS, perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS). There have been many studies linking PFOA and PFOS to certain health effects. There are, however, thousands of PFAS chemicals, and within this group there is a wide range of chemical structures with very different chemical and physical properties. The associated health effects for many of the different PFAS are unknown. Therefore, filling this knowledge gap is a top research priority for federal agencies.
PFOA and PFOS are often referred to as “long-chain” or “C-8” PFAS. While they are no longer manufactured in the U.S., historically, they were widely used directly or indirectly in the production of consumer products and PFOA and PFOS are persistent in the environment. For these types of PFAS, the FDA uses the EPA’s reference dose of 0.02 μg/kg bw/day as an appropriate toxicity reference value when conducting a safety assessment. The FDA does not currently have toxicity reference values for dietary exposure for PFAS other than PFOA and PFOS.
How is the FDA estimating the occurrence of PFAS in the general food supply?
To understand the occurrence of PFAS in foods, the FDA first had to develop reliable analytical methods to detect and measure these very complex chemicals in foods. In 2012, we began testing for certain types of PFAS in milk and later expanded testing to seafood and cranberries. In 2019, we were able to expand and validate the testing method with a diverse group of foods including breads, cakes, fruits, dairy, vegetables, meats, poultry, fish, and bottled water for 16 types of PFAS. We posted our validated method in October 2019.
The food samples the FDA is currently analyzing to understand the occurrence of PFAS in the general food supply are part of the foods collected for the Total Diet Study (TDS). The TDS is conducted on an on-going basis and serves as the FDA’s primary method of monitoring levels of various pesticide residues, contaminants, and nutrients in foods. PFAS are not currently part of the TDS. Results from the initial testing of PFAS in foods will be used to determine how the FDA will monitor PFAS in foods going forward, including whether steps should be taken to include it in the TDS, and/or if targeted sampling assignments are necessary for certain foods.
What has the testing for PFAS in foods shown so far?
As of December 2019, the FDA has conducted eight surveys designed to measure certain PFAS in foods generally and from specific areas with environmental contamination. Overall, we have found that very few foods have detectable levels of certain PFAS. From our recent surveys of foods that are part of the general food supply, the results of our first round of testing showed that out of 91 foods, two samples—ground turkey and tilapia—had detectable levels of one type of PFAS called PFOS. The PFOS levels that were measured in these samples were very low and are not likely a health concern. The second round of testing included 88 foods and showed that one sample—tilapia—had a detectable level of same type of PFAS. Again, the PFOS level found in the tilapia sample is very low and is not likely a health concern.
Publications from our previous work and data tables from our more recent surveys are available on the FDA’s PFAS webpage.
Do the results mean I should avoid tilapia or other seafood?
Currently, there is no scientific evidence that supports recommending consumers avoid a particular food, including tilapia or seafood. Other types of seafood were included in our limited testing, and they were not found to have detectable levels of PFAS. As part of a healthy eating pattern, fish and other protein-rich foods have nutrients that may offer health benefits for children and adults.
For recreational fish, the FDA recommends that you check the state’s fish consumption advisories for the specific area where you will be fishing. Fishing advisories may exist that provide recommended consumption rates of particular kinds of fish from particular water bodies where local contamination has occurred. If you do not know whether a water body that you fish in is covered under a fishing advisory, call your local or state health or environmental protection department.
A searchable list of fish advisory websites maintained by states, territories, and tribes is available on the U.S. Environmental Protection Agency’s website.
Why does the FDA allow PFAS to be applied to food contact surfaces, like non-stick pans?
PFAS are a very broad group of chemicals, and they act differently under different uses. This is why they are found in a wide variety of products, including everything from stain resistant clothing to firefighting foam, and also are used in some food contact applications. There are several different types of PFAS that the FDA has evaluated and are approved for use in food contact applications.
Some PFAS are approved for use in the manufacture of non-stick pan coatings. These coatings are made of molecules that are polymerized (i.e. joined together to form large molecules) and applied to the pan through a heating process that tightly binds the polymer coating to the pan, which prevents PFAS migration to food. Similarly, the PFAS used in manufacturing of gaskets that come into contact with food do not migrate to food or pose a safety risk because they are bound within the polymerized molecule.
The PFAS approved for use on paper or cardboard (to prevent grease from going through them) can potentially migrate to food. The FDA conducts a rigorous premarket safety review to ensure that the use of specific PFAS chemicals in food contact applications is safe. However, because of the growing scientific evidence about PFAS generally, the FDA is reviewing available scientific information and research to make certain their continued use in food contact applications is safe.
Does washing or cooking remove PFAS from foods?
PFAS cannot be removed from foods by washing or cooking. The levels of PFAS that have been found in foods from the general food supply, however, are very low and based on the best available current science, the FDA has no indication that these present a human health concern. | https://www.fda.gov/food/chemicals/questions-and-answers-and-polyfluoroalkyl-substances-pfas-food |
Sep 28, 2016
Share your input in this year’s Community Health Needs survey!
KSB Hospital’s mission statement calls for us to meet the healthcare needs of our community. The first step in that goal is to understand what the needs of the community are. You can click here to take a survey to help us understand our community’s needs. Every three years, we conduct a Community Health Needs Assessment, or CHNA, to gather input and set priorities for community health. This year, we’ve teamed up with the Lee County Health Department, who is conducting a similar study of local needs. We put together a diverse group of representatives from schools, law enforcement, health providers, advocacy groups, and community organizations to discuss the various needs around us.
You’re an important part of this process too. You can share your input on the quality and availability of local services, and suggest new ideas or priorities for KSB Hospital to pursue to improve local health.
Click here to take a brief survey and make your voice heard!
Thank you for your participation and for your continued support of healthy communities! | https://ksbpulse.com/2016/09/share-your-input-in-this-years-community-health-needs-survey/ |
Batik is beautiful patterned cloth that characterizes the richness of Indonesian culture, especially in Java such as Yogyakarta. Yogyakarta batik cloth that is versatile is actually a lot of functions. The development of batik as a fashion trend in various circles, whether young or old, to a variety of professions & economic background, more the emergence of modern batik motifs. One that often gets the spotlight is batik motif Yogyakarta or Jogjakarta. Batik Jogja or Batik Yogya is basically a batik that has a batik style with white base. Here is the long history of Batik Yogyakarta.
- Relatively closed with the Mataram Kingdom
The existence of Yogyakarta batik is closely related to the history of the establishment of the Islamic Mataram kingdom by Panembahan Senopati. After moving the royal center from Demak to Mataram, Panembahan Senopati often meditated along the coastal of Java Island, among others in Parangkusuma to Dlepih Parang Gupito, tracing a thousand mountainous cliffs that looked like “pereng” or cliff march.
As the King of Java who of course mastered the art, then the circumstances of the place inspired him in creating the pattern of batik slopes or machetes, which is a feature of agatar Mataram and different from previous batik patterns. Since the creator was the founding King of the Mataram Kingdom, then by his descendants, the machete’s patterns should only be worn by the King and his descendants in the palace.
Mataram Kingdom of Islam reached its heyday during the reign of Sultan Agung Prabu Hanyokrokusumo where the people live safely and prosperously. At that time the arts and culture also get high attention, one of which is batik, which has become a raw material and worn clothing in various opportunities. Based on some data obtained, the batik center at that time was in Pleret area.
Read also: History of Tugu Yogyakarta
- The Giyanti Agreemen
After Sultan Agung died and successively replaced by his heir, the circumstances changed and there was chaos. The number of conflicts then led to the Giyanti Treaty held on May 13, 1755, in which the Kingdom of Mataram split into two, they are Surakarta Hadiningrat Sultanate under Sri Sunan Pakubuwono III, the successor of Sri Sunan Pakubuwono II and Sultanate of Yogyakarta Hadiningrat Sultanate under Sri Sultan Hamengkubuwono I (Prince Mangkubumi).
The journey of “Batik Yogya” can’t be separated from the agreement of Giyanti 1755. As soon as Mataram is split in two, and Kraton Ngayogyakarta Hadiningrat stands, Mataram fashion is transported from Surakarta to Ngayogyakarta then Sri Susuhunan Pakubuwono II design new fashion and traditional dress of Kraton Surakarta different from Yogya fashion.
In the village of Giyanti, the talks took place. The results include, among others, the Region or Territory of Mataram divided into two, one part under the authority of Paduhunan II Paduhunan in Surakarta Hadiningrat, partly under the rule of Kanjeng Prince Mangkubumi who after being crowned as king with title Dalg Sampeyan Dalem Ingkang Sinuhun Kangjeng Sultan Hamengku Buwana Senopati ing Ngalaga Ngabdul Rachman Sayidin Panatagama Kalifatullah ingkang jumeneng kaping I, which is then called kratonnyaNgayogyakarta Hadiningrat.
All heirlooms and palace objects are also divided in two. Clothing Mataraman brought to Yogyakarta, because Kangjeng Prince Mangkubumi who intend to preserve it. Therefore Surakarta under the control of Sri Paduka Susuhunan PB III designed a new fashion arrangement and succeeded in making Surakarta Royal Palace as we have seen today.
Read also: History of Mix Martial Art in Indonesia
- Batik Yogya during 13th century
In the 13th century, the process of making batik is only done by the princesses and relatives of the King. Batik has become a skill that must be owned by the princess and family palace. For the family of Keraton, batik is a means to practice patience, perseverance as well as creativity, taste, and intention. The end result is also worn itself to meet the needs of traditional ceremonial clothing that is always held in the palace. Due to the high level of clothing needs in the Keraton environment, batik cloth work involving the community and businessmen outside the palace Keraton. The servants sometimes take home and do the process of making batik at home.
Read also: Javanese Batik Fabric
- Batik Yogya during 17th untill 19th century
In the 17th century to early 19th century batik survive to be an intermediary exchange in the archipelago. At that time batik which is the result of artwork of the palace became an interesting commodity trade along the northern coast. In royal palace of Yogyakarta, Javan Batik cloth is worn as a fashion. Batik cloth in the palace is a clothing outfit that are used for all purposes, daily clothing, clothing keprabon, fashion to attend the ceremony of tradition, etc. Javanese men’s clothing that consists of headgear, nyamping, campuh, all of them are made of batik cloth. So also with the completeness of the clothing of Javanese princess which is also in the form of batik cloth.
Read also: Batik in Indonesia
- The rule of wearing Batik in palace
Characteristic of Yogyakarta style batik, there are two kinds of background or fabric base color. They are white and black. The color of batik can be white (color of mori cloth), dark blue, and black brown. Sered (rim of fabric) is white. Both clothes are black and white. Simple decorative variety closely related to Hindu philosophy such as: Sawat symbolizes the crown or high ruler, Meru symbolizes the mountain or earth (earth), Dragon symbolizes water, Bird symbolizes the wind or the world over, Flame symbolizes fire. Since first there is already a ban fabric. Every Emperor of the Sultan is entitled to make new rules or prohibitions.
Sri Paduka Sultan HB VIII made a new regulation (revision) entitled Pranatan dalem Bab namanipun peangangge keprabon ing Nagari Ngayogyakarta Hadiningrat. The Batik prohibition motif is Parang rusak (parang rusak barong and parang rusak gendreh).
Read also: Clothes to wear in Bali
- Batik Yogyakarta Today
Batik Yogyakarta including batik hinterland or batik is very different from batik pesisiran character. Characteristic of Yogyakarta style batik has two kinds of background or the basic color of cloth that is white and black.
Along with its development, batik more widely used community outside the Palace of Yogyakarta and appears certain groups such as Pamong class batik, batik Sudagaran, and batik public. Batik class Pamong widely used by the pamong who came from the Palace. There are also among those who obtain the title of nobility for meritorious in the field of culture and government. Not surprisingly, the among certain levels, often seen wearing batik style Keratonan. They are often involved in events and ceremonies in the palace. | https://factsofindonesia.com/history-of-batik-yogyakarta |
Will the non-human recruiters please stand up?
In this week's edition, we bring to fore some of the ways different companies in India are already leveraging AI in HR processes and are not only cutting down on manual labor but also seeing massive savings, and in cases even higher revenues.
If you were born before the '70s, you'd clearly remember the protests against the introduction of computers in banks. Protestors would go on strikes in a fight to safeguard the interests of the employees of public sector banks.
Despite the concerns, technology has so far been proven to be a net employment generator than otherwise, and therefore when automation hit the recruitment industry itself, there have been more cheers than boos. Recruiters today have realized that AI and other automation technologies can supplement their work instead of competing against them. From resume filtering to validating the potential candidate's certificates, there are a whole lot of activities that are part of a recruiter's daily life that isn't all sunshine and roses. AI helps cut down on these boring yet necessary activities while allowing recruiters to focus on more meaningful work.
In this week's edition, we bring to fore some of the ways different companies in India are already leveraging AI in HR processes and are not only cutting down on manual labor but also seeing massive savings, and in cases even higher revenues.
Do write to us with your feedback.
Regards Varun Aggarwal Editor, ETCIO [email protected]
Making AI do all your grunt work
When the companies grow at a higher rate, it becomes nearly impossible for humans to scan thousands of CVs and profiles to select just the right candidates. So, the companies have started automating their systems to scan these CVs and find the right talent from the market.
“AI makes it possible to efficiently screen a far wider selection of candidates, increasing the likelihood of finding scarce talent in today’s talent wars. Specifically, it helps in the process of shortlisting out of a large number of applicants bringing some transparency and consistency of the decision-making process, reducing scope for mistakes and biases. There is an increasing use of AI based assistants for writing more effective job postings and recruiting emails as well,” said Prashanth Kaddi, Partner, Deloitte India. Read more...
The majority of hiring in CSS Corp happens through resume scanning which flows in the system through various sources. To go through those resumes, the company has developed a cognitive search integrated with the internal system that identifies the right candidate. CSS Corp calls this system TAMS (Talent Acquisition Management System).
TAMS identifies the right resumes which flow into the systems and matches them to the resource requirements and then presents the top 10 resumes that the team can look at and call for interviews. This is being done with Microsoft Cognitive search and integration with other backend systems. Read more...
The AI based approach to Talent lifecycle especially on the acquisition side plays a major role in the Recruitment effectiveness, productivity as well as Talent experience. This enables not only Higher Velocity and faster scaling of teams but also ensures optimization of the time spent by the engineering teams in shortlisting the right talent thereby improving their motivation and productivity.
"While the initial Models resulted in the reduction of Resume perusal volumes by 30% by way of match between the job description and the potential candidate's skill set, the effectiveness and fitment of the identified repository has gone up by 50% thereby enhancing pipeline to selection ratio significantly. While this is the initial bootstrap model training period, we expect the model efficiency to go higher and impact ramp up velocity between 30 - 40% enabling a much faster onboarding of teams," V. Padmanabhan, Vice President – Engineering, GlobalLogic explained. Read more...
How automation helped this recruitment firm nearly double its revenue
When the candidates submit their applications for a job posting, they are automatically sent the pre-screening tests. These are on three levels. First is the technical abilities, second is psychometric test and the third is competency mapping respect to the various KRAs of job description. Based on the results of these tests and the applications made, the system intelligently screens and filters the candidates for recruiters to interview.
“It is a one time investment and it is surely worth investing in. We were able to break even in a matter of a few months. More than reducing the hiring cost for us, it has helped us generate more revenue. A very important factor to consider here is ROI. The time that our recruiters saved with the automated and intelligent system was used to close multiple other high-value positions . Since our team of recruiters were not getting filtered profiles, they gave more time and focus on closing high value positions which lead to an increase in revenue by 70%,” Suvarna Ghosh, Founding Partner at Maxima Group explained. Read more... | |
We believe that a firm compliance setup will benefit both our customers’ and our own business performances by streamlining processes covering Quality, Health & Safety and Environment (QHSE).
At DSV, we offer our customers the same high compliance standards across markets based on our global Integrated Management System.
We empower the management in each country to decide if there’s a need for them to be ISO certified. We support this through our global ISO multisite certification which controls key management functions with maximum efficiency and minimum bureaucracy while reducing the overall cost for our customers.
Our firm compliance setup enables us to gather comprehensive data throughout the value chain covering processes within Quality (ISO 9001), Health & Safety (ISO 45001) and Environment (ISO 14001). This way, the compliance framework offers a solid fact-based decision-making tool to support continual improvements, increased efficiencies, reduction in errors and improved overall business performance.
The framework creates value by generating:
- Operating efficiency: DSV can reduce costs by following industry best-practice and by focusing on quality.
- Improved decision-making: DSV can detect and identify problems in due time, which means that DSV can quickly take steps to avoid the same mistakes in the future.
- Process integration: By examining process interactions, DSV can improve efficiency, reduce errors and our customers can benefit from cost savings.
- Continual improvement: Continual improvement is a core principle of the ISO standards. We utilise the framework as a systematic approach to identifying and exploiting opportunities to improve.
- Credibility: The ISO standards are internationally agreed upon by experts and considered a formula that describes the best practice to ensuring high quality in our operations.
Any questions? | https://www.dsv.com/en/sustainability-esg/governance/policies/management-systems |
APPSeCONNECT is an emerging software product in the Integration Platform as a Service (iPaaS) industry and through its Omni-channel integration capabilities, it empowers businesses to leverage their existing systems, resources, technology, and data. It seamlessly streamlines business operations, thus increasing productivity and efficiency.
This simple, yet powerful tool enables synchronizing best critical business applications like eCommerce, ERP, CRM, POS, etc. It empowers businesses to automate their vital business processes, eliminating the need for any painful manual data exchange by enabling real-time, bi-directional data exchange between them. APPSeCONNECT has been successful in streamlining the processes and accelerating the growth of over 500 businesses across 40+ countries. | https://oroinc.com/b2b-ecommerce/partner/appseconnect |
In the context of my work, in addition to the existing racism and white supremacy, class distinctions which lead to issues of poverty, result in divisions within the society. My peace education work helps me to be more effective in organizing interactive engagements with others around these issues. I have increased my communication and pedagogical skills and expanded the methodologies I use in facilitating and organizing gatherings.
|Start in the field & motivations / Inicio en este campo y motivaciones|
I started working in the field of peace education due to my commitment to social justice and non-violence. I was also inspired by people, whom I liked and respected, who were actively working in peace education.
|Significant Career Moments & Success Stories / Momentos significativos de tu carrera y historias de éxito|
The experience of understanding, appreciating and valuing the dialectic of connecting and being alike, on the one hand, and being different on the other hand, which comes to light in relationships with specific individuals at peace education gatherings, have been noteworthy moments in my peace education work. | https://people-pe.ahdr.i-i-p-e.org/people/entry/74/ |
Blog
Blog
Whose personal is more political: Women who use drugs and the feminist movement
3/8/2019 7:37:00 AM |
By Judy Chang, IAS Member and Executive Director of the International Network of People Who Use Drugs
“In a marketplace of experiences, the privileged inevitably have more platforms from which to narrate, and the marginalised are often spoken for within agendas which are not their own.”
Alison Phipps 2016, p. 5-6, Sussex University
I am a feminist. I am a woman who uses drugs. Up until recently, these identities have been mutually exclusive, having rarely been held together in the same conceptual space.
Only now are the links between drug policy, feminism and drug use beginning to be drawn. Feminism, I took to early on, as any woman who questioned dominant paradigms, interrogated inequities in political and social life and was sceptical of the ways gendered norms condition how we speak, act, behave and interact with one another.
Twenty years ago, university life introduced me to the feminist movement. Grounded in radical politics, it was liberating, insightful and welcoming of diverse communities and identities, including women of colour, indigenous women, lesbian, bisexual, queer and trans* women and women living with disabilities. The only exceptions, at the time, were female sex workers and women who use drugs.
Twenty years ago, I also started injecting heroin: the part of myself I hid and suppressed.
Whilst diversity was embraced in feminism, particularly for those who faced intersecting oppressions, drug use and sex work seemed to provoke profound discomfort in the women’s movement. Women who use drugs were, at best, invisible, but more often than not, pitied and/or loathed and rendered irrational, useless and hopeless. On the mere basis of our consumption patterns, we were somehow to blame for our own oppression, bereft of self-awareness and incapable of knowing our own minds and controlling impulses, and therefore undeserving of participation in the feminist movement.
Women who use drugs issues are feminist issues. Due to criminalization, punitive policies and stigma and discrimination, women who use drugs are two to five times more likely to experience gender-based violence and intimate partner violence compared with women who don’t use drugs (El-Bassel et al. 2011, Gilbert et al. 2015, Moore 2008).
It is not just the violence itself that is the problem, but that it, more often than not, is perpetrated with impunity. Gender inequality means that we face higher mortality rates and HIV and hepatitis C prevalence compared with men (Larney et al. 2015, Esmaeili et al. 2017). Our sexual and reproductive health rights are suspended, where we are at times subject to forced abortion and sterilization, routinely denied child custody rights, turned away from health services and treated as second-class citizens. Harsh drug laws coupled with patriarchal norms and assumptions have led to the mass incarceration of women who inject and use drugs. In many parts of the world, women who use drugs are more likely than men to be incarcerated for the same offence or face harsher sentencing laws. Due to the double, sometimes triple, stigma faced by women who use drugs, the rights violators are not held accountable. With all of this in mind, it is troubling that empathetic links of solidarity have not been extended by many mainstream feminists to women who use drugs.
Kimberlé Crenshaw’s 1989 theory of intersectional feminism is a useful framework to examine the overlapping systems of oppression and discrimination women face, based not just on gender and sex, but on race and ethnicity, sexuality, economic background and other status and other axes, including drug use. As much as it helps us to think through differences, it can also help in considering the commonalities between systems of oppression, in this case, patriarchy and drug prohibition. Both of these are predicated on the same objectives and principles, the suppression and subjugation of difference, the control of bodies, limitations on personal choice and freedom and the silencing of dissenting voices.
In such a society, women are necessarily judged more harshly for their drug use than men. We are judged more harshly than men are for taking personal risks, for being cast as putting our own needs and the pursuit of pleasure above “caring for others”.
Twenty years later, I have seen the feminist movement be changed thanks to the advocacy of sex workers and transwomen. Just like feminism was challenged and inevitably changed as a result of the agendas of second-wave feminism of the ’60s-’80s and third-wave feminism, it must continue to confront racism, classism, sexism, heterosexism, transphobia and able-bodyism, as well as whore phobia and drug user phobia.
We’re no longer willing to be marginalized by a movement that we hold claim to, where our bodies are deemed impure or our behaviour unnatural or immoral. What, after all, could be more patriarchal than telling a woman what to do with/put in her body?
Women who use drugs need our space and voices heard. There is movement; one of NarcoFeminism. We are coming together, caring for each other, mobilizing and organizing, coming out at conferences to demand self-determination and empowerment. For feminism to move forward, it must resist and challenge outdated norms that are incompatible with real-world realities.
If the claim of feminism is to stand up for women, it must be to stand up for all women.
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Schools are going to re-open for the academic year 2021-2022 with classroom learning and optional distance learning. We thought this could be a time to shed some light on engaging kids in the learning process.
If you are a teacher and wish to create a better interactive experience with your students, and wish them to engage more in learning, you should be following these steps. The idea is to recruit their interest, which means finding ways to make learning “relevant, authentic, and valuable” in students’ lives.
In this post, we are going to talk about 5 steps you can follow to actively engage your students and help them feel personally connected to their learning.
- Connect things with real-life examples.
The best way to engage students in their learning is to make sure the material speaks to them.
Use teaching materials that are culturally relevant. Students who don’t find representations of their own cultures in texts are likely to lose interest in school-based literacies.
Have your students complete a short survey on their outside interests and use that information to assist in building your lesson plans. This will help your students see the connections between what they are learning inside and outside the classroom.
Another easy way to help students feel personally connected to what they are being taught is to talk about how they can apply the material in real life. Teachers demonstrate how students can apply math concepts to help them manage their personal finances, nutrition, and daily schedule.
Link routines to learning. You can also promote learning through classroom routines. For instance, a child learning to wash hands during bathroom breaks can also be taught science concepts (body parts, hygiene and disease prevention, water conservation), reading (bathroom signage), antonyms (hot/cold, left/right), and math (counting).
- Use students’ interests and fascinations.
Find out what your students are passionate about and then use those interests as natural motivators to increase engagement. Whether a child is fixated on one thing or has a few areas of intense interest, there are many simple strategies you can use to work those fascinations into your instruction. The result? Happier, more motivated students.
Allow a child to integrate their most-loved characters and possessions into your classroom reading time. One student was able to participate in a reading circle when his turn came once he was allowed to speak through a favorite puppet.
If you’re working on a math lesson, consider asking a student to write a problem, diagram, or pattern that relates to her particular area of interest. Sometimes, the best way to combine academic material with a student’s interests may not be immediately evident—but your students may see connections that you don’t!
You can explore a range of Sinarline Exercise books here you can use while trying to engage your kids in various learning activities:
https://sinarline.com/product-category/school-stationery/
- Give students choices
Be it doing homework or working on classroom assignments, make sure you give them options to select. When students get choices they feel they are making a decision for themselves and ultimately become more responsible in completing their tasks.
- Hook their interest with fun transitions.
Research says, “All students are more engaged when they enjoy classroom life, laugh, and connect with peers.” Transitions between activities can be the perfect time to infuse more joy and fun into your daily routine–and get your students energized and excited to learn.
- Teach students self-monitoring skills
An advanced way of involving children so they stay engaged in their learning is to help them develop greater self-regulation skills. Children sometimes struggle with self-awareness, so they may not even realize when they’re straying off task or acting in disruptive ways. When children are taught to regulate their behavior and work independently, they develop habits to help them succeed and you are freed to operate more flexibly in the classroom.
When you make a concerted effort to engage students in their learning, they’ll be better able to maintain focus, sustain positive behavior, and grasp and retain the material you’re working so hard to deliver—a positive outcome for everybody! | https://sinarline.com/how-to-get-students-more-engaged-in-learning/ |
Why is my no-bake cheesecake gooey?
Trying to use a baked crust (even one with less butter) means your filling will ooze into the crust and make it soggy. Try this instead: Most importantly, you should make sure to chill the crumb crust at least 10 minutes before filling it, but even longer is better. This makes for a sturdy crust when refrigerated.
Do no-bake cheesecakes taste good?
Whereas a baked cheesecake has eggs in the recipe, then the cheesecake is baked in a water bath, then it chills in the fridge. The textures of these two cheesecakes are totally different. The no-bake version is a lot softer and almost mousse-like. Both are absolutely delicious.
What happens if no-bake cheesecake doesn’t set?
After the chill time, if your cheesecake still isn’t firm enough, you can freeze it to help it set up. Be careful not to freeze it too long unwrapped though; it can cause the texture to become gritty.
How do you keep a no-bake cheesecake from melting?
In most cases, the best way that you can fix your recipe for a no-bake cheesecake (assuming that the recipe you are following is not faulty), is adding gelatin to the cheesecake. Gelatin is a natural thickener and it is used in many recipes to help thicken sauces, custards, and other foods.
How can I speed up my no-bake cheesecake?
How to Thicken No-Bake Cheesecake
- As mentioned, the most important step is to beat cold heavy cream into peaks. Fold the whipped cream into the cheesecake filling gently so you don’t deflate the air.
- Refrigerate the cheesecake for at least 6-8 hours, but overnight is better.
Does no-bake cheesecake contain gelatin?
A no-bake cheesecake is a light, creamy version of a cheesecake that requires no cooking at all. Cream cheese, heavy cream, gelatin, and vanilla come together for a filling that firms up in the refrigerator.
Which is better no-bake cheesecake or baked cheesecake?
The prime difference between no-bake cheese and baked cheesecake is that the baked one has a better texture. In particular, baked cheesecake has a creamy and smooth texture which is hard to achieve with a no-bake cheesecake recipe. The baked cheesecake tends to be fluffy and moist even if it’s baked.
Why is my cheesecake like mousse?
The baked cheesecakes have a fairly dense, sometimes “squeaky” texture as they have been set with a combination of eggs and heat. The cheesecakes that are set with gelatine may have a fairly smooth texture or a very mousse-like texture but will not be dense.
Can you put cheesecake in freezer to set faster?
For this reason, you need to let the cheesecake come down to room temperature before you put it in the freezer. Also, you can put the cheesecake in the freezer for a night or a minimum of four hours to ensure it solidifies perfectly. Place the cheesecake on the stand and place the plastic wrap over the cake’s top.
What happens if I don’t put gelatin in cheesecake?
Third, I also add gelatin to this cheesecake to help it firm up and make it easy to slice. Without it, the cheesecake is fairly soft and will gradually deflate.
How to make the best eggnog cake recipe?
Preheat oven to 350º Fahrenheit and spray a 9″ square inch pan with cooking spray.
Does it taste like eggnog?
In fact, a glass of eggnog tastes like the farthest thing from morning eggs or a warm dinner meal — a glass of eggnog actually literally tastes like melted ice cream in a glass that’s somehow hugging your throat as you drink it. There really is no equivalent to eggnog — it’s one-of-a-kind. Will There Be A Third Wave Of Coronavirus In The UK?
What is eggnog recipe?
“Whip cream until stiff. Add 1/4 cup sugar. In another bowl, whip egg whites and 1/2 cup sugar until stiff. In a third larger bowl, whip egg yolks until creamy with 1/2 cup sugar. Add liquor to whipped egg yolk mixture. Stir in milk. Mix all ingredients together. Chill before serving.
Is eggnog a custard?
Yes, eggnog can taste very much like custard. It depends on what the person fancies in their mug of eggnog. And it depends on what ingredients are omitted. If one wants to cut the volume of alcohol in their serve of eggnog, then it can easily pass as a custard. | https://www.wazeesupperclub.com/why-is-my-no-bake-cheesecake-gooey/ |
Browse by Subject "Theatre"
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Attention, absorption and habit: the Stanislavski System reexamined as a cognitive process using the "Theatre of Consciousness" model of Bernard Baars (2010-05-19)Konstantin Stanislavski stated that “Art should be on good terms with science.” This was far from an unqualified statement, as he stridently resisted the “scientific sophistries” that some actors used to replace true ...
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Bound (2014-05)Adaptation thrives in our entertainment culture and as consumers we encounter adapted works regularly; yet still we cannot seem to escape the fidelity criticism that birthed such phrases as the book was better. I am drawn ...
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(2014-05-30)Grounded in the historiography of the field of trauma studies and a theoretical understanding of the role of adaptation within contemporary culture, this dissertation examines how the process of adapting the familiar ...
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(2012-09-18)Herbert Blau is more widely known in theatre history as a theorist and less as a director who strove to identify his directorial style during a time when American values were evolving and American regional theatres were ...
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(2011-08-26)The relationship between William Shakespeare’s black Othello and white Desdemona has held a particular hold on the American imagination for two-hundred and fifty years. During its history on the New York stage, Shakespeare’s ...
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(Im)possibilities of building community and negotiating belonging in institutional theatre during the wars in Croatia and Bosnia: a comparative study of cases from Sarajevo, Zagreb, and Belgrade (2015-07-15)This dissertation examines how and to what end institutional theatre participates in the process of building community and negotiating belonging from 1991 to 1995 during two major conflicts of the Yugoslav Succession Wars: ...
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(2012-05-22)The Ludus de Antichristo, composed c. 1157-1160 at the imperial monastery of Tegernsee, Bavaria is one of the most complete dramatic texts to have survived the Middle Ages. I use the term “complete” because it can be ...
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(2015-07-17)This dissertation analyzes the performance methodology and reception of various flash mobs performed from 2003 to 2015. This twelve year history is separated into three periods. The first period consists of Bill Wasik’s ...
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(2014-05)With this image I wanted to connect the three plays that are discussed and compared in my research pertaining to new play development. The clock seen in the image is distorted with symbols representing each play respectively ...
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(2016-04)As a MFA candidate in Theatre Design, I recently had the privilege to sound design Not About Nightingales by Tennessee Williams. I was confident in all the music specified by the playwright except for one lyrical piece of ... | https://www.ideals.illinois.edu/browse?type=subject&value=Theatre |
Builder claimed 24 bricks cost $1k in loss
A MAN who took 24 bricks from a vacant block at the Capricorn Coast believed they were rubbish from nearby construction sites.
Craig Rex Rodgers, 58, pleaded guilty on April 17 in Rockhampton Magistrates Court to one count of stealing.
Police prosecutor acting Sergeant Ellysha Geddes said Rodgers was photographed and filmed removing cement building bricks from the site and loading them into his Ford Falcon ute.
She said Department of Main Roads were able to identify the owner of the ute for police.
Sgt Geddes said police located the bricks at Rodgers home when they went to speak to him about the incident.
She said Rodgers admitted to taking the bricks, saying two unknown males at the property told him he could remove them.
The court heard Rodgers admitted to taking 24 bricks, but did not agree with the restitution amount of $1000 provided by Chris Warren Homes without an itemised quote.
Defence lawyer Brian McGowran said his client, who was on a disability pension due to an incident that left him in a wheelchair until he learned to walk again, had been a bricklayer prior to his accident.
He said Rodgers had gone to the construction sites to talk with Chris Warren about a job, but was denied the work due to the seriousness of his back injury.
Mr McGowran said the vacant block of land were the bricks were taken from looked like a dumping ground for excess material from the construction.
"He wasn't taking it off pallets. He did it in broad daylight," he said.
Mr McGowran said Boral sells the bricks for $2.50 per brick and Bunnings sold them for $4 per brick.
"The estimate of $1000 seems to have been plucked out of thin air," he said.
Mr McGowran said Rodgers offered to buy the bricks off the company, but police said the matter had to go through the courts. Magistrate Philippa Beckinsale ordered Rodgers to pay restitution of $300 and fined him $500. A conviction was recorded. | https://m.themorningbulletin.com.au/news/builder-claimed-24-bricks-cost-1k-in-loss/3998878/ |
Classics is the generic term for the study of the Greek and Roman languages, history, society, culture, religion and archaeology. No other subject area spans such a wide range of different and interesting elements, from literature to philosophy, from Linear A and Linear B to Greek and Latin, from temple architecture and vase painting to burial mounds, from matricide to suicide, from Celts to Celtiberians, from Picts to Parthians; in fact, you name it and Classics will have it covered somewhere down the line.
Classical Civilisation is taught at GCSE and A Level and is a non-linguistic subject that requires no knowledge at all of Latin or Greek; just a love of the world of the Greeks and Romans.
No one can deny the influence of the Greeks and Romans on world civilisation even today; the ancient Greek idea of democracy continues to be regarded as the ideal form of government; the Roman senate lives on in American politics; theatre is still a very popular career and art form; and Latin continues to feature prominently in the language of lawyers and doctors and is the preferred language for use in mottoes and on coinage; it is also the root language for French, Italian, Portuguese, Romanian and Spanish, and will give you a real head start when learning them.
Latin and Classical Civilisation obviously go well with each other at A level [yes, you can study both], but they also complement English, History, Mathematics, MFL, Politics, Philosophy and Science.
University degrees in classical subjects are highly regarded and students with a good honours degree in Classics can expect to be a hot commodity and in great demand by employers. | https://www.mgs.kent.sch.uk/information/departments/subjects/classics/ |
Should everyone have access to health care in a language they can understand? This is not a new question. In 2001, Health Canada released a report entitled ‘Language Barriers in Access to Health Care,’ calling for further research, including an exploration of the feasibility and cost of providing interpretation throughout the health care system. Since then, while interpretation programs have been expanded in some jurisdictions in Ontario, we are no closer to guaranteeing that every patient receives health care in a language they can understand. In fact, the conversations, policies and funding related to interpretation programs in Ontario largely seem to position it as a ‘nice to have,’ opt-in bonus, rather than an essential component of provider/patient interactions.
In terms of policy in Ontario, the current grab bag of approaches suggests there are no clear, binding provincial guidelines regarding health care language accessibility. Studies suggest, however, that when providers and patients can’t communicate in the same language – sometimes termed ‘language discordance’ – this results in poorer health outcomes, and even increased risk of death. A 2002 study of 1,800 people with active tuberculosis born outside of Canada and living in Ontario found an association between language discordance and increased risk of death. A 2004 study of three Toronto hospitals found that language discordance led to increased length of hospital stays, and a 1990 study of 22,448 women in Ontario suggested that women whose primary language was not English were less likely to receive some forms of cancer screening.
We also know there are many jurisdictions in Ontario where patients who do not speak English will be unable to find a primary care provider who speaks their language. A recent study from the Centre for Research on Inner City Health (CRICH) found several municipalities in need of Portuguese, Italian, Punjabi, Chinese and Spanish-speaking primary care physicians, although in some cases the population in need was small. Currently, CRICH is conducting an analysis for the Toronto Central Local Health Integration Network (TC LHIN) looking at the degree to which patients are able access care from a primary care physician who speaks their language in different areas of Toronto. The study is ongoing, but preliminary results suggest there are large gaps in different areas of the city.
When interpretation is offered, we see a clear improvement in the health care experience. In 2014, CRICH conducted an evaluation of the TC LHIN’s telephone interpretation program, Language Service Toronto (LST). Established in 2012, LST currently offers 25 participating hospitals and 18 community health organizations in or near central Toronto on-demand access to telephone interpreters speaking 170 languages, 24 hours a day, 7 days a week. Interpreters are available to health care workers for appointments and follow-up. They are also available to administrative workers for appointment booking and other interactions. Our evaluation found that relatively seamless access to telephone interpretation both improved the health care experience and allowed organizations to reach people they were not able to previously.
The positive impact of the right type of interpretation at the right time is clear. So is the fact that language discordance between health care organizations and patients can lead to negative impacts on health outcomes, and potentially unnecessary additional use of health care service (ie. lengthier hospital stays). While LST Toronto is an excellent start, it is a voluntary program. Organizations opt in – hospital and community-based health organizations are not required to offer professional interpretation. In addition, we don’t know the degree to which independent primary care providers, who are often the first point of contact to the health care system, are using professional interpretation. Even within organizations subscribed to LST, use may be uneven. It is unclear, for example, whether all providers (physicians, nurses, social workers, case coordinators, etc.) and support staff (receptionists) are trained on and offered the use of LST within every organization.
Access to telephone interpretation is one component of what could be a coordinated, multi-pronged approach to ensuring language concordance between health care organizations and patients in Ontario. Our evaluation of LST found that telephone interpretation should be supplemented with other forms of interpretation including in-person, adaptations for people who are hard-of-hearing, and video conferencing (for detailed findings, please visit www.crich.ca/reports). A previous study suggests additional strategies including increased recruitment of medical students who speak needed languages and a publicly-available database of physicians who speak a range of languages. Finally, when obtaining a license in Ontario, international medical graduates are required to practice in ‘underserved areas.’ The definition of ‘underserved’ could be broadened to include areas that require physicians who speak non-official languages.
There is no doubt that a coordinated health care language accessibility strategy is needed in Ontario and, likely, at a national level. The stated objective of the Canada Health Act is to ‘facilitate reasonable access to health services without financial or other barriers.’ We would argue that language discordance is a barrier that needs to be addressed, not based on the goodwill of individual providers and organizations, but at all levels of the health care system, and for everyone.
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Within the cultural mosaic of Canada, with diverse population speaking different languages and practicing various cultures and traditions, the value of effective communication cannot be underestimated.
In medical practice, history taking is the cornerstone of patient care which forms the basis of proper examination and precision of diagnosis. So it is not difficult to predict the consequences of language discordance with increased length of hospital stay, inaccurate or missed diagnosis, ineffectiveness of health promotion campaigns are just a few to name.
Thanks for sharing this valuable research and I strongly support your point that provision of language services should not be an optional service but should be made available at all levels to provide efficient and equitable healthcare to all.
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Well stated. | https://healthydebate.ca/opinions/everyone-access-health-care-language-can-understand/ |
Bowei Xi - Purdue University
Students: Spring 2022, unless noted otherwise, sessions will be virtual on Zoom.
A Game Theoretic Approach for Adversarial Machine Learning -- When Big Data Meets Cyber Security
Feb 27, 2019Download: MP4 Video Size: 230.4MB
Watch on YouTube
AbstractNowadays more and more data are gathered for detecting and
preventing cyber attacks. Unique to the cyber security
applications, learning models face active adversaries that try to
deceive learning models and avoid being detected. Hence future
datasets and the training data no longer follow the same
distribution. The existence of such adversarial samples
motivates the development of robust and resilient adversarial
learning techniques. Game theory offers a suitable framework to
model the conflict between adversaries and defender. We develop a
game theoretic framework to model the sequential actions of the
adversaries and the defender, allowing players to maximize their
own utilities. For supervised learning tasks, our adversarial
support vector machine has a conservative decision boundary,
whereas our robust deep neural network plays a random strategy
inspired by the mixed equilibrium strategy. One the other hand,
in real practice, labeling the data instances often requires
costly and time-consuming human expertise and becomes a
significant bottleneck. We develop a novel grid based adversarial
clustering algorithm, to understand adversaries' behavior from a
large number of unlabeled instances. Our adversarial clustering
algorithm is able to identify the normal regions inside mixed
clusters, and to draw defensive walls around the center of the normal
objects utilizing game theoretic ideas. Our algorithm also
identifies sub-clusters of adversarial samples and the overlapping areas
within mixed clusters, and identify outliers which may be
potential anomalies. | https://www.cerias.purdue.edu/news_and_events/events/security_seminar/details/index/6tpbk57s26lsgdjt1sgr7sscrg |
Main Areas of Research
- One aspect emphasized by my studies was analysis of the various call types, primarily the advertisement calls; beginning with anurans native to Germany, the project was then extended to many species in other regions of Europe and beyond.
- Because anurans are ectothermic, many of their bodily functions are influenced by the ambient temperature, including the calls. Hence one objective was to determine the extent to which important characteristics of the mating calls and other call types are correlated with the temperature. Knowledge of these changes is indispensable in order to draw conclusions about the species-specificity of the advertisement calls. Furthermore, it is a crucial prerequisite for comparing the calls of the different species.
- Whereas the behaviour of other animal groups, for instance the birds, has long been the subject of intensive research, until very recently the behaviour of native anurans had attracted no appreciable interest. But the anurans have proved to be distinguished by diverse and specific behavioural traits that mainly determine their calling and reproductive behaviour. Hence another point of emphasis was to clarify these traits, e.g. by monitoring the daily and seasonal variations in the calls of male frogs and demonstrating the precise responses of the females to conspecific and simulated mating calls.
- Comparative morphological investigations revealed the structure of the larynx, the vocal cords and the laryngeal muscles, as well as other muscles involved in call production. Electron microscopic and experimental studies clarified the fine structure and the physiological properties of the laryngeal musculature.
- Physiological analyses provided information about the arrangement of the rest of the vocal system, the nerve centres involved in calling and the interaction of various neural structures during call production.
- Because the advertisement calls are extremely specific characteristics of a species, they constitute a valuable criterion for establishing the kinship relations among various anurans. Bioacoustic comparisons proved successful in tree frogs and water frogs, and resulted in a reorganization of anuran systematics, including the discovery of new species.
- At a later stage in the bioacoustic studies the anuran auditory organs were also included, as well as the sense organs in the labyrinth that mediate perception of gravity and movements. Comparative anatomical experiments provided information about the structure of the labyrinth in many anuran species, as well as the number and arrangement of the sensory cells at the sensory end points.
- The fine structure of the sensory cells themselves and the nature of stimulus transmission were an additional point of emphasis, as well as the development of the labyrinth and the differentiation of the sensory epitheliums during embryonic and larval development.
- Physiological studies in several native anuran species taught us about the neuronal auditory threshold curves and how they change under the influence of temperature. | http://dr-hans-schneider.de/schwerpunkte_e.html |
China agriculture and forestry carbon (01069) extension letter of intent, involving the acquisition - Business News - Qingdao Hanil Precision Industry Co., Ltd.
Chinese forestry low carbon (01069) released, reference is made to the company in October 31, 2016 may be related to Shenzhen in the acquisition of housing assets announcement of the container box.
As stated in the announcement, the company entered into a letter of intent with the Shenzhen Chinese container on potential acquisitions. According to the letter of intent, the letter of intent will automatically expire if there is no formal agreement on potential acquisitions within 3 months from the date of the letter of intent.
The Shenzhen case also promises not to negotiate or negotiate with any other third parties in any form for the sale of the target business from the date of the letter of intent to the date of signing the formal agreement.
On the date of the announcement, the parties to the letter are still discussing and discussing potential acquisitions. Therefore, in January 26, 2017 after the closing, the company and Shenzhen signed letters of intent to postpone the box expiration date postponed, accordingly, if the company and the Shenzhen in the box is not on or before April 30, 2017 to enter into a formal agreement of intent will automatically expire. | http://en.specialcontainer.com/news_view.aspx?typeid=5&id=443&fid=t2:5:2 |
What are common diseases of respiratory system?
Respiratory system diseases. Diseases and conditions of the respiratory system fall into two categories: Infections, such as influenza, bacterial pneumonia and enterovirus respiratory virus, and chronic diseases, such as asthma and chronic obstructive pulmonary disease (COPD).
What is the most common disease in the respiratory system?
The most common lung diseases include:
- Asthma.
- Collapse of part or all of the lung (pneumothorax or atelectasis)
- Swelling and inflammation in the main passages (bronchial tubes) that carry air to the lungs (bronchitis)
- COPD (chronic obstructive pulmonary disease)
- Lung cancer.
- Lung infection (pneumonia)
What are the disease of circulatory and respiratory system?
Emphysema (COPD) – Chronic Obstructive Pulmonary Disease, of which emphysema is one of, results in progressive destruction of the air sacs in the lungs and loss of respiratory membrane for oxygen exchange. The bane of long term smokers. Atelectasis – a collapsed lung.
How do you treat respiratory disease?
Pharmacological Treatment of Respiratory Disorders
- Bronchodilators. These are the most frequently used inhaled medications.
- Corticosteroids.
- Mast Cell Stabilizers and Anti-IgE Antibodies.
- Leukotriene Receptor Antagonists.
- Antihistamines and Epinephrine.
- Pulmonary Surfactants.
- Antimicrobials and Antivirals.
- Other Agents.
What are the 5 diseases of the respiratory system?
The Top 8 Respiratory Illnesses and Diseases
- Asthma.
- Chronic Obstructive Pulmonary Disease (COPD)
- Chronic Bronchitis.
- Emphysema.
- Lung Cancer.
- Cystic Fibrosis/Bronchiectasis.
- Pneumonia.
- Pleural Effusion.
How can I check my lungs at home?
How It Is Done
- Set the pointer on the gauge of the peak flow meter to 0 (zero) or the lowest number on the meter.
- Attach the mouthpiece to the peak flow meter.
- Stand up to allow yourself to take a deep breath.
- Take a deep breath in.
- Breathe out as hard and as fast as you can using a huff.
- Note the value on the gauge. | https://englewoodcardiacsurgery.com/diseases/diseases-of-the-respiratory-system.html |
The modern era in art, stretching roughly from the mid-18th to the mid-20th century, could be said to comprise a long series of bold statements, each aimed at carrying artistic practice into uncharted territories. As industrialization created a new middle class and an urbanized society, artists responded to the changing environment with new ways of seeing and rendering.
At Heather James Fine Art in Palm Desert, the exhibition Masters of Modernism and Impressionism surveys many of the movements that made the avant-garde so exciting. The robust show of approximately 25 works features examples from the Impressionist, Post-Impressionist, Fauvist, and Cubist eras, among others — an opportunity to view a span of historical avant-garde works side by side.
The short-lived but highly influential Fauvist movement built on the achievements of the Impressionists by foregoing naturalistic renditions in favor of intensely vivid, high-contrast colors. The Heather James exhibition includes a striking work by Kees van Dongen, a Dutch artist who became a core member of the Paris-based Fauves. Van Dongen loved the glamour of Parisian life and was known for his sometimes-racy portraits of fashionable society women. Recuerdo de Toledo (1908), with its wistful title and apparent setting on the outskirts of a Spanish town, is a moving example of the artist’s work with nudes, featuring subtle manipulations of green and yellow highlights against a rich blue background.
While the great Fernand Léger is considered a major figure in the Cubist movement, the actual trajectory of his career was multifaceted, absorbing influences from Fauvism, futurism, Surrealism, neo-classicism, and social realism. The breadth of his oeuvre shows he was a fiercely independent artist who consistently hewed to his own vision. La racine noire et fragment d’objets (The black root and fragments of objects), dated between 1943 and 1950, was painted close to the end of his life, yet it shows no decline in the rigor of his practice. An abstract work that references organic and man made objects, the painting marries the flat planes of Cubism with the free association of Surrealism.
Heather James Fine Art, which also shows contemporary artists, appears intent on making an annual tradition of bringing Old Masters exhibitions to the Coachella Valley; the gallery published a book documenting last year’s Picasso exhibition. As gallery curator Chip Tom sees it, the Old Masters are not only a popular draw in their own right, but also they shed useful light on the practices of artists working today.
As if to prove this point, Masters of Modernism and Impressionism opened in November along-side Evocatecture, an intriguing group show of contemporary artists who use architectural forms as a lens through which to view psycho-logical states, cultural mores, and political frameworks. The five featured artists include the German-born Stefan Kürten, whose densely layered paintings investigate the failed dreams of the bourgeois world, and Naomi Safran-Hon, an expatriate Israeli artist who explores the ways in which landscape constructs identity in her conflict-ridden home country. These contemporary struggles offer a fascinating complement to the formal innovations of previous eras.
Masters of Modernism and Impressionism continues through April 30 at;
Heather James Fine Art
45188 Portola Ave. | https://www.palmspringslife.com/surveying-the-masters/ |
Transcranial direct current stimulation (tDCS) is a widely-used tool to induce neuroplasticity and modulate cortical function by applying weak direct current over the scalp. In this review, we first introduce the underlying mechanism of action, the brief history from discovery to clinical scientific research, electrode positioning and montages, and parameter setup of tDCS. Then, we review tDCS application in clinical samples including people with drug addiction, major depression disorder, Alzheimer's disease, as well as in children. This review covers the typical characteristics and the underlying neural mechanisms of tDCS treatment in such studies. This is followed by a discussion of safety, especially when the current intensity is increased or the stimulation duration is prolonged. Given such concerns, we provide detailed suggestions regarding safety procedures for tDCS operation. Lastly, future research directions are discussed. They include foci on the development of multi-tech combination with tDCS such as with TMS and fMRI; long-term behavioral and morphological changes; possible applications in other research domains, and more animal research to deepen the understanding of the biological and physiological mechanisms of tDCS stimulation.
National Natural Science Foundation of China(31400959)
; 973 Program(2014CB846102)
; 111 Project(B07008)
; Open Research Fund of the National Key Laboratory of Cognitive Neuroscience and Learning(CNLZD1306)
; Fundamental Research Funds for Central Universities(SWU1509422
; Open Research Fund of the Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences(KLMH2015G01)
; Research Program Funds of the Collaborative Innovation Center of Assessment toward Basic Education Quality at Beijing Normal University(2016-06-014-BZK01
; 15XDSKD004)
; SCSM-2016A2-15003)
| |
Motor vehicle crashes remain the leading cause of death and acquired disability in children.1 The American Academy of Pediatrics (AAP)2 and the National Highway Traffic Safety Administration (NHTSA)3 recommend appropriate restraint of children in motor vehicles on the basis of child age and size to reduce injury risk. Children younger than 1 year or weighing <20 lb should ride rear-facing in child safety seats (CSSs). If a CSS accommodates a child rear-facing to higher weights, then for optimal protection, the child should remain rear-facing to the maximum weight of the car safety seat as long as the top of the head is below the seat back. Once children have outgrown their rear-facing CSS, they should be restrained in a forward-facing CSS until the upper weight limit (typically 4 years of age and 40 lb) for the child restraint. Children who have outgrown their CSS should be restrained in a belt-positioning booster seat (BPB) with 3-point seat belt until the seat belt fits properly (∼80 lb and 57 inches in height, typically at least 8 years of age).
Although seat belts are better than no restraint at all, appropriate child restraint has demonstrated effectiveness in reducing injury risk when compared with restraint by seat belts alone. Recent evidence demonstrates that CSSs reduce the risk of severe, life-threatening injury by 71% when compared with seat belt restraint alone for children between the ages of 12 and 47 months.4 BPBs reduce the risk of head and brain injuries, all internal organ injuries, spinal cord injuries, and extremity fractures by 59% when compared with seat belt restraint in 4- to 7-year-old children.5 In particular, BPBs virtually eliminated seat belt syndrome in these children; that is, injuries to the abdominal organs and lumbar spine and spinal cord.
In 2000, the NHTSA launched a new wave of outreach efforts by multiple groups focused on appropriate restraint for children over age 4.10 The primary foci of these efforts were improved access to BPBs, educational and media campaigns, and upgraded laws. At the time of this article’s writing, 22 states (Arkansas, California, Colorado, Delaware, Illinois, Louisiana, Maine, Maryland, Montana, Nebraska, Nevada, New Hampshire, New Jersey, Oregon, Pennsylvania, Rhode Island, South Carolina, Tennessee, Vermont, Virginia, Washington, and Wyoming) and the District of Columbia had passed upgraded child restraint laws that require the use of booster seats for children over age 4.11 Many other states are actively considering similar upgrades to their child restraint laws so that they are in line with best practice recommendations. In light of this recent outreach activity, the objective of this study was to examine trends in restraint type used among children under age 9 to evaluate the success of these efforts in improving appropriate child restraint usage in older children while maintaining realized benefits in appropriate restraint of the youngest children in 3 large regions of the United States.
Data were collected from December 1, 1998, to November 30, 2002. A description of the study methods has been published previously.12 The project consists of a large-scale, child-specific crash surveillance system: insurance claims from State Farm Insurance Corporation (Bloomington, IL) function as the source of subjects, with telephone survey and on-site crash investigations serving as the primary sources of data. The study protocol was reviewed and approved by the Institutional Review Boards of both the Children’s Hospital of Philadelphia and the University of Pennsylvania School of Medicine.
Vehicles that qualified for inclusion were State Farm insured and involved in a crash with at least 1 child occupant under age 16. Only vehicles from model year 1990 and later were studied to focus on current vehicle safety-design features. Qualifying crashes were limited to those that occurred in 15 states and the District of Columbia, representing 3 large regions of the United States (East: New York, New Jersey, Pennsylvania, Delaware, Maryland, Virginia, West Virginia, North Carolina, District of Columbia; Midwest: Ohio, Michigan, Indiana, Illinois; West: California, Nevada, Arizona). After policyholders consented to participate in the study, limited data were transferred electronically to researchers at the Children’s Hospital of Philadelphia and the University of Pennsylvania. Data in this initial transfer included contact information for the insured, the ages and genders of all child occupants, and a coded variable describing the level of medical treatment received by all child occupants as reported by the policyholder.
A stratified cluster sample was designed to select vehicles (the unit of sampling) for the conduct of a telephone interview with the driver. Vehicles that contained children who received medical treatment after the crash were oversampled so that the majority of injured children would be selected while maintaining the representativeness of the overall population. When a vehicle was sampled, all child occupants in that vehicle were included in the survey. Drivers of sampled vehicles were contacted by telephone and, if a passenger had received medical treatment, screened via an abbreviated survey to verify the presence of at least 1 child occupant with an injury. All vehicles with at least 1 child who screened positive for injury and a 10% random sample of vehicles in which all child occupants who were reported to receive medical treatment but screened negative for injury were selected for a full interview; a 2.5% sample of crashes for which no medical treatment was received were also selected. Claim representatives correctly identified 95% of eligible vehicles, and 74% of policyholders consented for participation in this study. Of these, 19% were sampled for interview and an estimated 81% of these were interviewed successfully. The full interview involved a 30-minute telephone survey with the driver of the vehicle and parent(s) of the involved children. Only adult drivers and parents were interviewed. The median length of time between the date of the crash and the completion of the interview was 6 days, with 95% of interviews completed within 47 days of the crash.
The eligible study population consisted of all 10 195 children who were younger than 9 years and riding in 8730 State Farm–insured vehicles that were 1990 or newer and whose driver reported a crash claim between December 1, 1998, and November 30, 2002. Complete data on crash date and child’s age were acquired for the study sample. The percentage of missing data on restraint type was no more than 1%.
Restraint status of children was determined from a series of questions in the telephone survey that probed descriptions of the restraint type and how it was used. Children were classified as unrestrained or restrained, with the restraint type further classified as CSS (either forward- or rear-facing), BPB, shield booster seat (ShB), and seat belt. Rear-facing CSS included infant seats designed for rear facing only (many of these seats have a handle to carry the seat) and rear-facing convertible seats (these seats are designed to be used either in a rear- or forward-facing orientation). Forward-facing CSSs included forward-facing convertible seats and forward-facing combination seats used with the internal harness (these seats are used with an internal harness until the child reaches the manufacturer’s maximum weight limit and then are converted to a BPB by removing the internal harness). BPBs included high-back and backless booster seats and those combination seats used without the internal harness. ShBs are seats that are installed in the vehicle with the lap portion of the adult seat belt and have a padded shield, as opposed to a harness, to keep the child in place. The definition of seat belt included lap only, lap/shoulder, and shoulder only.
Among the 161 children for whom paired information on restraint use was available from both the telephone survey and crash investigations, agreement was 88% between the driver report and the crash investigator (κ = .74, P < .0001).
The primary purpose of these analyses was to examine trends in restraint use by the total group and 6 age groups (0-2, 3, 4, 5, 6, and 7-8) from December 1, 1998, to November 30, 2002. The beginning of the study period is defined as its first 6 months (December 1, 1998, to May 31, 1999), and the end of the study period is defined as its last 6 months (June 1, 2002, to November 30, 2002). Because sampling was based on the likelihood of an injury, subjects who were least likely to be injured were underrepresented in the study sample in a manner potentially associated with the predictors of interest.13 To account for this potential bias and to adjust for the clustering of subjects by vehicle, we used analytical methods to account for sampling weights, sampling strata, and sampling units. The sample weights, defined as the inverse of the inclusion probability of the child in the sample, account for the differential sampling rates. Weighted data then reflect estimates for the entire population in an approximately unbiased manner. Because of the complex sample design, we used SAS-callable SUDAAN: Software for the Statistical Analysis of Correlated Data, Version 8.0 (Research Triangle Institute, Research Triangle Park, NC) for all data analyses. Descriptive statistics, consisting of frequencies for categorical variables, were determined. To describe the relationship of restraint use and time in a functional form, we modeled the log-odds of a given restraint type used (vs other) as a linear function of time via logistic regression, both using the total sample and restricted to each of the 6 age groups. The secular trends in restraint type use are then reported and plotted as the predicted probabilities resulting from this logistic regression, overall and for each age group. The dependent variables examined were CSS use, ShB use, BPB use, and seat belt use. Eight 6-month periods (from December 1, 1998, to November 30, 2002) were used as the independent variable to assess the time effect. Wald χ 2 statistical test was performed for the significance of the time effect.
Table 1 displays the distribution of restraint type by total and age group and 2 time periods (the first 6 months and the last 6 months of the study period). The distribution of restraint type varies by age group and time period, and very few young children were unrestrained.
Overall, at the end of the study period, 63% of children in this age group were restrained in a child restraint (CSS, ShB, or BPB); however, 62% of children aged 4 to 8 remain inappropriately restrained in adult seat belts. The probability of CSS use in the entire population increased significantly from 0.40 to 0.52, and the probability of BPB use also increased significantly from 0.03 to 0.11 (both P ≤ .001). The probability of ShB use decreased significantly from 0.06 to 0.02, and the probability of seat belt use decreased significantly from 0.51 to 0.34 (both P ≤ .001).
At the end of the study period, 98% of children in this age group were restrained in a child restraint. The probability of CSS use, ShB use, and seat belt use remained stable for this age group. The probability of BPB use showed a small but statistically significant increase (P = .028). BPB use increased significantly from 0.004 to 0.02 over the time period of study.
Figure 1 shows the trends in restraint type used by 3-year-olds. At the end of the study period, 95% of children in this age group were restrained in a child restraint. There was no significant trend in the probability of BPB use. CSS use increased significantly from 0.41 to 0.85 (P ≤ .001). In contrast, the probability of ShB use decreased from 0.30 to 0.03 (P ≤ .001). Concurrently, the probability of seat belt use decreased from 0.20 to 0.06 (P = .003).
Trends in restraint type used by children age 3 between 1998 and 2002.
Figure 2 shows the trends in restraint type used in this age group. At the end of the study period, 74% of children aged 4 were restrained in a child restraint. The probability of ShB use showed no significant trend, whereas the probability of CSS use showed a highly statistically significant increase from 0.19 to 0.48 (P ≤ .001). The probability of BPB use also increased from 0.10 to 0.22 (P = .03). Conversely, the probability of seat belt use showed a highly statistically significant decreasing trend from 0.60 to 0.23 (P ≤ .001).
Trends in restraint type used by children age 4 between 1998 and 2002.
Figure 3 (the CSS trend and BPB trend lines are virtually identical; for separating these 2 lines, 0.5% was added to the intercept for calculating the probability of BPB use) shows the trends in restraint type used by 5-year-olds. At the end of the study period, 48% of children in this age group were restrained in a child restraint. The probability of ShB use showed no significant trend, whereas the probability of CSS use increased from 0.03 to 0.25 (P = .002). The probability of BPB use also showed a highly statistically significant increase from 0.03 to 0.25 (P ≤ .001). Conversely, the probability of seat belt use showed a highly statistically significant decrease from 0.89 to 0.44 (P ≤ .001).
Trends in restraint type used by children age 5 between 1998 and 2002. The CSS trend and BPB trend lines are virtually identical. For separating these 2 lines, 0.5 was added to the intercept for calculating the probability of BPB use.
At the end of the study period, 29% of children in this age group were restrained in a child restraint. The probability of CSS use increased from 0.01 to 0.09 (P = .012). The probability of BPB use also showed a highly statistically significant increase from 0.01 to 0.25 (P ≤ .001). In contrast, the probability of ShB use showed a decreasing trend from 0.02 to 0.002 (P = .07). Concurrently, the probability of seat belt use decreased from 0.94 to 0.68 (P ≤ .001).
At the end of the study period, 5% of children in this age group were restrained in a child restraint. Trends in ShB use could not be analyzed because of the small number (n = 3) over the study period. The probability of CSS and BPB uses both showed a small increases in use from <0.01 to 0.03 and <0.01 to 0.05, respectively (P = .03 for CSS; P = .043 for BPB). In contrast, the probability of seat belt use showed a small but statistically significant decrease from 0.97 to 0.90 (P = .016).
Our research findings demonstrate dramatic shifts in the distribution of restraint types used by US children under age 9 over a short period of time between 1998 and 2002. Overall, there was a 29% increase (from 49% to 63%) in child restraint system use for children under age 9 with a concomitant 27% decrease in seat belt use. At the beginning of the study, approximately half (49%) of children in the study population were inappropriately restrained by seat belts, and approximately half of them (49%) were in child restraints. By the end of the study, the majority of children in the study population (63%) were in child restraints, and 36% of them used seat belts. Among children aged 4 to 8, however, more than half (62%) remained inappropriately restrained in seat belts.
In our crash-based study, for children under age 9, the prevalence of serious injury remained stable over the period of the study (P = .13): from a low of 1.3% (unweighted n = 132) for the first half year to 1.4% (unweighted n = 131) for the last half year. Despite the increased use of child restraint for these young children, the prevalence of serious injury remained essentially unchanged within this short period of time. Because the cause of injury for children in crashes is multifactorial,14 this stability might be the result of the change of other factors or the high seat belt usage for children aged 4 to 8 over the study period. This might further indicate the need for continued efforts and promotions for booster seat use for children in this age group.
Other studies have demonstrated increased use of child restraints by US children. In 2001, National Safe Kids17 reported that, among children aged 4 to 8 observed in their convenience sample, 37% were estimated to use a booster seat. Compared with 27% of booster seat use among children aged 4 to 8 observed in a convenience sample in 1999, this increase is substantial.18 According to an observational study conducted by the NHTSA18 on the use of child restraints in 2002, for children aged 1 to 3, the use of forward-facing child safety seats increased to 62% in 2002 from 39% in 2000. Our study supports and extends these results by describing trends in specific restraint types used for each age of child. Our methods highlighted that the greatest gains were demonstrated for children aged 3 and that although there was an increase noted overall in booster seat use, use of specific types of boosters varied. Despite gains in appropriate restraint, a great proportion of older children, especially the 7- and 8-year-olds, were still inappropriately using the vehicle seat belt as their restraint. This points to the need for continued and improved intervention efforts that promote appropriate restraint for children aged 4 to 8.
Qualitative studies19,20 identified lack of knowledge about the importance of appropriate restraint and the risks associated with premature use of seat belts as major barriers for parents to restrain their 4- to 8-year-old children appropriately. The most important barrier identified was the lack of concordance between state laws and best practice recommendations. In addition, as emphasized in their planning for a national strategy to increase booster seat use for children aged 4 to 8, the NHTSA10 proposed addressing challenges such as “the lack of information among parents or caregivers,” “young child’s desire to act grown up and not to have to sit in any type of child restraint,” and “inconsistency of state laws pertaining to protecting older children and booster seat use.” Sustained outreach efforts and upgrades to state child restraint laws are important in addressing these barriers or challenges.
The increasing trend in CSS use among children over age 4 deserves attention. Before 2000, few CSSs were rated by manufacturers to hold children who weigh >40 lb.23 The promotion of appropriate restraint for children under 9 had an impact on the design of the new CSSs. Several of the new CSS makes can hold children up to 50 lb,16,24 yet few of these seats are likely in our study. Our data showed a dramatic increase in the CSS use by 4- to 5-year-old children. The emphasis on appropriate child restraint use for older children had a significant impact on child passenger protection in the United States. Many parents delayed graduation of children to seat belts until older ages. As an added benefit, many more children stayed in CSSs through age 4, with a substantial minority continuing through age 5. Educational efforts aimed at promoting booster seat use by children over age 4 seemed to “raise the bar” for all children, reducing premature graduation out of CSSs and into seat belts. The risks and benefits of harness restraint by children who weigh >40 lb should be evaluated.
At the current time, federal legislation is pending to encourage states to upgrade their child restraint laws to mandate the use of booster seats for children over age 3 (Cathy Chase, Advocates for Highway and Auto Safety, personal communication; www.saferoads.org; 2003). Our results may assist state and federal policymakers by demonstrating the current level of interest on the part of parents to restrain their older children appropriately.
These results reflect very current trends in child restraint usage. Although considerable achievements have been realized over a short period of time, substantial inappropriate restraint still remains: 62% of children aged 4 to 8 remain inappropriately restrained in adult seat belts. Parents hear safety messages when they are relevant to their children. As a result, sustained efforts about appropriate restraint must continue to maintain and improve the gains achieved in appropriate child restraint use. The additional benefits realized by recent changes in child restraint laws remain to be evaluated.
We thank State Farm Insurance Companies for their financial support of this work through the Partners for Child Passenger Safety project. In addition, we thank the many dedicated claim representatives and personnel from State Farm, the Research Team on the Partners project and at TraumaLink who devoted countless hours to this study, and the parents who generously agreed to participate in the study. | https://pediatrics.aappublications.org/node/42386.full.print |
A balance of flexible and inflexible qualities make Haskell a fascinating programming language to learn and use.
First, the Haskell programming language is not named after Eddie Haskell, the sneaky double-dealing neighbor kid in the ancient TV sitcom, Leave It To Beaver.
Haskell is named after Haskell Brooks Curry, an American mathematician and logician. If you don't know, logicians create models to describe and define human reasoning, for example, problems in mathematics, computer science, and philosophy. Haskell’s main work was in combinatory logic, a notation designed to eliminate the need for variables in mathematical logic. Combinatory logic captures many key features of computation and, as a result, is useful in computer science. Haskell has three programming languages named after him: Haskell, Brooks, and Curry.
Haskell the language is built around functions, useful blocks of code that do specific tasks. They are called and used only when needed.
Another interesting feature of functional languages like Haskell: functions are treated as values like integers (numbers) and strings. You can add a function to another function the way you can add an integer to an integer, 1 + 1 or 35 + 53. Perhaps the best way to describe this quality is a spreadsheet: in a cell in the spreadsheet, you can add numbers as well as a combination of functions to work on numbers. For example, you might specify each number in cells 1-10 be added up as a sum. In Excel, at least, you also can use SUMIF to look for a pattern in cells 1-10 and, if the pattern is found, perform an action on any cells with the pattern.
What Makes Haskell Special?
Technically, Haskell is a general-purpose functional programming language with non-strict semantics and strong static typing. The primary control construct is the function. (Say that fast ten times!) Here's what it means:
- Every language has a strategy to evaluate when to process the input arguments used in a call to a function. The simplest strategy is to evaluate the input arguments passed then run the function with the arguments. Non-strict semantics means the input arguments are not evaluated unless the arguments passed into the function are used to evaluate what is in the body of the function.
- Programming languages have rules to assign properties — called a type — to the components of the language: variables, functions, expressions, and modules. A type is a general description of possible values the variable, function, expression, or module can store. Typing helps minimize bugs, for example, when a calculation uses a string ("house” or "cat”) instead of a number (2 or 3). Strong static typing evaluates the code before runtime, when the code is static and possibly as code is written.
- The order in which statements, instructions and functions are evaluated and executed determines the results of any piece of code. Control constructs define the order of evaluation. Constructs use an initial keyword to flag the type of control structure used. Initial keywords might be "if” or "do” or "loop” while final keywords might be "end if” or "enddo” or "end loop”. Instead of a final keyword, Haskell uses indentation level (tabs) or curly brackets, or a mix, to indicate the end of a control structure.
Perhaps what makes Haskell special is how coders have to think when they use the language. Functional programming languages work in very different ways than imperative languages where the coder manages many low-level details of what happens in their code and when. While it is true all languages have things in common, it’s also true languages are mostly functional or mostly imperative, the way people are mostly right handed or left handed. Except functional programming languages require a different way of thinking about software as you code.
Other features that make Haskell interesting:
- Strong data typing (evaluating properties of all inputs into a function) is combined with polymorphism; a function to sort numbers also can be used to sort strings of text. In some languages, you would have to code two or more functions, one for each data type.
- Lazy evaluation (one of my favorite coding terms!) allows the result of one function/task to be handed to another function/task on the same line of code. For example, the command can search a file for all instances of a string then pass the results to be printed to the computer screen. Functions that can take other functions as arguments or return them as results also are called higher order functions.
- No side effects. In other languages, code can affect the state of the computer and application, for example, writing to a file. Haskell strictly limits these side effects which, in turn, makes Haskell applications less prone to errors.
- Haskell uses monads, a structure that works like an assembly line where every stop on the line performs a different task. This allows Haskell to separate side effects as a distinct activity apart from any function, for example, logging any errors as a function performs tasks on its data inputs.
Building from small bits of code, each bit tightly contained and testable.
How is Haskell Used?
As a functional programming language, Haskell has benefits like shorter development time, cleaner code, and high reliability. The tight control of side effects also eliminates many unforeseen interactions within a code base. These features are especially of interest to companies who must build software with high fault tolerances, for example, defense industries, finance, telecommunications, and aerospace.
However, Haskell also is used in web startups where functional programming might work better than imperative programming. Apparently Facebook, Google, NVIDIA, and other companies use Haskell to build internal tools used in their software development and IT environments. Even a lawn mower manufacturer in Kansas uses Haskell to build and distribute their mowers. And the New York Times recently used Haskell to build an image processing tool for the 2013 New York Fashion week.
The main Haskell website also has links to university courses, as well as research groups.
Learn More
Try Haskell
Haskell
http://www.haskell.org/haskellwiki/Introduction
http://www.haskell.org/haskellwiki/Why_Haskell_matters
http://www.haskell.org/haskellwiki/Functional_programming
http://www.haskell.org/haskellwiki/Category:Theoretical_foundations
http://www.haskell.org/haskellwiki/Lambda_calculus
http://www.haskell.org/haskellwiki/Haskell_in_industry
http://www.haskell.org/haskellwiki/Haskell_in_research
http://www.haskell.org/haskellwiki/Haskell_in_education
Haskell Brooks Curry
http://en.wikipedia.org/wiki/Haskell_Curry
Combinatory Logic
http://en.wikipedia.org/wiki/Combinatory_logic
Monads
http://en.wikipedia.org/wiki/Monad_%28functional_programming%29
Functional Programming and Imperative Programming
https://en.wikipedia.org/wiki/Functional_programming
https://en.wikipedia.org/wiki/Declarative_programming
https://en.wikipedia.org/wiki/Imperative_language
Side Effects
https://en.wikipedia.org/wiki/Side_effect_%28computer_science%29
Also In The November 2013 Issue
My Adventures with Raspberry Pi
Open source hardware geared towards artists, hobbyists, designers, and students, is a viable and far less expensive alternative to build your own computers.
Beth Rosenberg Talks How Tech Kids Unlimited Helps Kids Who Learn Differently
With a wave of kids with special needs graduating high school, how can technology help them with resumes, college, jobs, and careers?
Stop Words
A clever technique to speed up database searches also is an interesting concept.
More Fun with Raspberry Pi
Here are some videos, and links to even more videos, to learn how to use your Raspberry Pi and have all kinds of fun with Pi projects.
My goal wasn’t to make a ton of money. It was to build good computers.
News Wire Stories for November 2013
Interesting stories about computer science, software programming, and technology for the month of October 2013. More stories can be found at the Software Programming and Computer Science News Wire link at the top of every page of this site.
Bubble Sorts
With a bubble sort, numbers sort themselves as they bubble to the left of a group of numbers. Here's a fun catchy video to explain.
The 7 Bridges of Königsberg
This month's math puzzle dates back to 1735 when it was first solved by Leonhard Euler, a Swiss mathematician and physicist.
Pair Programming
From the start of computing history, people have tried to optimize the software programming process. This includes having two coders work together to code software.
Learn More Links for November 2013
Links from the bottom of all the November 2013 articles, collected in one place for you to print, share, or bookmark.
Haskell
A balance of flexible and inflexible qualities make Haskell a fascinating programming language to learn and use.
Icon-itis
The release this fall of Apple's iOS7 operating system is a great opportunity to explore the history of computer interface design.
Functions
Managing inputs and outputs is a key problem programming languages face. Here's how a few languages use functions to manage and transform data. | https://www.kidscodecs.com/haskell/ |
The gameplay of Heavenly Sword resembles a martial arts title focused on melee combat while featuring opportunities for ranged attacks. The main character, Nariko, uses a weapon called the "Heavenly Sword" which changes into one of three forms depending on what attack stance the player uses as part of a unique fighting style. Speed Stance provides an even balance between damage and speed, where the sword takes the form of two separate blades. Range Stance allows fast, long-range, but weaker attacks, with the sword being two blades chained together. Power Stance is the most powerful, but slowest style, where attacks are made with the Sword in the shape of one large, two-handed blade.
For exploration and certain battles, the game also makes use of "quick time events" (QTE). During a QTE, a symbol for a certain button or for an action such as moving the analog stick to the right or left appears on screen and the player must match what is shown to successfully complete the scene.
In addition to Nariko, a secondary character, Kai, is controlled for some portions of the game. Many of Kai's stages take the form of sniping missions, using her crossbow to pick off enemies, in some cases to protect characters. While Kai cannot perform hand-to-hand combat, in stages that call for her to explore the level she is able to hop over objects and to free herself from an enemy's grasp by temporarily stunning them.
Projectiles can be maneuvered to their targets using the motion-sensing capabilities of the SIXAXIS controller through a feature known as Aftertouch. Such projectiles include guiding Kai's arrows after she has launched them, and for Nariko, guiding a cannon or rocket launcher, or picking up and throwing objects.
GUIDES
FAQ
Stuck and need some help? Check out some of the questions the community have asked relating to this game. If none of these answer your problem or you want to get some tips when it comes to a particular level, feel free to submit a question of your own. | https://nerdburglars.net/videogame/heavenly-sword/ |
A frequently asked question by founders at the end of a pitch meeting is, “What does your process look like?”. This is another way of asking:
a) When can I expect you to tell me whether they are interested in my company?
b) How much time should I expect to dedicate to the process?
While no two are the same, I will sketch out the typical investment-decision process at NXTP Ventures when we are leading an investment round. This should give founders a better understanding of what goes on behind closed doors.
The Initial Pitch Meeting
This meeting is all about getting to know the founders and understanding whether their idea fits with what we look for in a startup.
- Do the founders seem sharp, driven, and analytical?
- Are they addressing a large market opportunity?
- Are they meeting that opportunity with a well-thought-out business model?
Sometimes, we don’t have the answer to all those questions after a 45-minute meeting but we will have a hunch. And if it is positive we will openly express that we are eager to spend time improving our understanding of the opportunity. If we don’t have a firm point of view, we will likely communicate that we need time to discuss internally. Usually, we will revert in less than a week with feedback or information on next steps.
Initial Assessment of Opportunity
If the pitch sparks interest, we start the process with few work streams that run in parallel:
- Desk work: This consists of researching the market, product, and competitors.
- Founder reference checks: We reach out to our network to learn more about the founders. We speak to everybody from former colleagues and bosses, to mentors, friends, and acquaintances.
- One-page summary: We put together a one-page summary on the opportunity for discussion with the investment team.
- Expert network: Once the team agrees we should spend more time on the deal, we usually reach out to our expert network for a validation of the opportunity and continue with founder reference calls.
Our investment process builds incrementally towards the conviction there is a real opportunity to build a large business. This is an iterative process. We are gathering as many data points as possible so that, if and when we say yes, we do so with maximum conviction and enthusiasm.
Second Call with Founders
After our initial desk work and expert calls, we put together a list of questions pointing to where we see potential challenges for the business and areas we would like to learn about. Our main objective for second meetings is to learn more whether it meets what we look for in a startup and to answer a series of questions about the founding team:
- How are the founders responding to and thinking through the questions we ask them? We try to assess the depth of the founding team’s answers and see how analytical and data-driven the responses are.
- Have they already thought about these questions before and have they have performed any analysis on them?
- Does the company show a data-driven culture? While founders answer questions, it is great to see them open live, working documents and dashboards that the company uses internally. This gives us a glimpse into company culture and how they approach key challenges.
Oftentimes, these calls will end with a first data request such as a spreadsheet file with that contain the following information:
- Historic company KPIs
- Revenue per customer file
- Information on customer acquisition and retention
- Go-to-market strategy
Shipping these documents promptly is appreciated and, data in hand, we dig into the numbers to learn more about the business and to answer questions about the financial profile of the business, revenue growth & upsell, customer acquisition & retention, and gross margins, to name a few. All the while, founder reference calls continue and should be well above five at this point.
Meeting the Rest of the Team
As we process the data, we continue with expert calls and produce a new list of questions, more detailed and specific. At this point, we like to invite other members of the management team to the meetings to get an idea of the quality of the C-suite executives. We love seeing a deep bench of quality professionals as this is often indicative of the type of talent the organization is capable of attracting. We typically meet with the head of marketing or sales, the head of product, the CFO, CTO, or any other key members of the team.
Once these meetings conclude, we hope to have substantially concluded our business diligence. Pre-COVID, these meetings typically took place in person during two-day on-site visits. That is our strong preference. However, we are being forced to get comfortable with doing this remotely nowadays.
Getting to a Term Sheet
With the business diligence largely completed and if all reference calls check out, we proceed by sending the founders an email summarizing the terms of the proposed investment. Once we align on the macro terms, we get the lawyers involved to draft a formal term sheet. If there are any outstanding items that we still need to diligence, we will lay them out or include them as conditions precedents to the signing of the final financial agreements.
With a signed term sheet we kick off legal due diligence and, depending on the stage of the company, financial due diligence. These processes are run by qualified third parties that work closely with you to make sure the company is investment-ready and we have a full understanding of its legal and financial history.
Signing a term sheet is a huge commitment for us and it should be interpreted as such. It means we are committed and excited to partner with you and, barring any seriously troubling new information or a stark deterioration in business fundamentals, we will
Final Financing Agreements
As the final legal due diligence (and financial DD for later-stage companies) is conducted, we usually get the ball rolling on drafting the final financing documents. Ceteris paribus, this process will be more time-consuming for later-stage companies and more straightforward for early-stage startups.
For seed rounds, we usually opt for minimizing cost and time by running with the standard series seed investment agreement or opt for SAFEs/convertible debt agreements. We like to spend as much time as necessary going over the terms with founders to make sure they have a full understanding of what they are signing.
Throughout the process we have frank conversations with the founders about what it means to work with us, what we expect of them, and what they can expect of us. Setting clear expectations is key to a successful and enjoyable working partnership.
Length of Process
The final question to answer is: How long does this process take? Unfortunately there is no one answer as this depends on a variety of factors. From first meeting to term sheet could be as little as three weeks up to three months. Usually, once a term sheet is signed, final investment docs are countersigned approximately four weeks later, though this can sometimes be shorter or longer depending on the underlying structure and complexity of the legal and financial due diligence.
Does this sound like an involved process? Perhaps. But picking an investor for your company should not be a decision you take lightly. Due diligence processes are a two-way street. Founders should welcome the opportunity to get smarter on how investors view their business and, more importantly, get to know the partners they are going to work with for the next five-to-ten years. | https://nxtp.vc/2020/06/17/our-investment-process/ |
Your tasks:
- Be a part of the Project Team that is responsible for setting up an efficient, digital and successful German mortgage product
- Responsible for designing and implementing end-to-end new mortgage product operations & processes
- Manage policies, procedures to ensure compliance with various regulations and requirements
- Focal point for handling and providing guidance on a variety of escalated issues and complex resolutions, and questions from the team
- Ensure defined process & procedures are being followed in day-to-day mortgage operations
- Review & monitor overall operational & service performance with sufficient metrics, develop quality control processes to secure continuous improvement
- Build vendor relationship management strategy and work with departments & vendors to agree and drive the SLAs for service delivery to secure continuous improvement, respond to business needs & emerging changes over time
- Capture training needs, implement necessary trainings, create and maintain operational procedure & processes
- Translate operational requirements into our scope, user stories and work with the Team to formulate clear acceptance criteria and deliver our backlog
Your knowledge/experience:
- Several years of experience in operations preferably within a regulated banking environment
- Thorough understanding of aspects of mortgage lending, servicing and operational policies & practices
- Extensive knowledge of mortgage related regulations, policies & underwriting standards
- Experience in managing vendor performance monitoring and vendor relationship
- Strong collaboration and influence skills to effectively lead cross-functional alignment
- Proven German and English communication skills
We offer:
- An international team with colleagues from more than 25 different countries
- An excellent learning culture and opportunity to improve your professional competencies
- Different possibilities to flexibly organise your working hours, e.g. part time, flex time
- Possibility to work from home at any time – also after Covid-19
- Social benefits such as a company pension scheme, life insurance and capital formation contributions
- 30 days holiday as well as 24 and 31 December as non-working days
- Additional benefits such as employer contributions to BVG job ticket, fruit and salad days, free sports activities
- Easily accessible city centre offices, in the vicinity of many shops and restaurants to which you receive a discount
Bank of Scotland is committed to equal opportunities and actively promotes diversity and equality. We therefore welcome applications from candidates with disabilities. This position is available on a part time or full time basis.
We look forward to receiving your complete application (max. 3 MB) with a letter of motivation, CV, references, salary expectations and earliest possible start. | https://www.bankofscotland.de/karriere/stellenausschreibung-operations-manager-mortgages |
Download Small Business and Entrepreneurship Class 11 notes PDF and score well in the exam. These Notes are prepared by our expert teachers at cbsencertsolutions. Class 11 Small Business and Entrepreneurship Notes assist you with overviewing the chapter in minutes. At exam time, Revision note is one of the best tips suggested by educators during exam times.
These Small Business and Entrepreneurship Class 11 Important Questions are compulsory for the Class 11 Board Exam. Small Business and Entrepreneurship Class 11 notes are exceptionally useful to revise the entire syllabus during exam time. These notes cover all significant topics and Concepts given in the section.
Please refer to Small Business and Entrepreneurship Class 11 Business Studies notes and questions with solutions below. These revision notes and important examination questions have been prepared based on the latest Business Studies books for Class 11. You can go through the questions and solutions below which will help you to get better marks in your examinations.
Class 11 Business Studies Small Business and Entrepreneurship Notes and Questions
SMALL BUSINESS
Small scale industries contribute significantly to the development process and acts as a vital link in industrialization in terms of production, employment and exports for economic prosperity by widening the entrepreneurial base and use of local raw materials and indigenous skills.
Types of Small Business
Business enterprises are classified as Manufacturing Enterprises, Service Enterprises,Village Industries and Cottage Industries. Among these manufacturing and service enterprises
are again subdivided into Micro, Small and Medium Enterprises. The definition used by the Government of India to describe small industries is based on the investment in plant and machinery. The Micro, Small and Medium Enterprises Development (MSMED) Act 2006 has been passed by the Government of India to address the issues of small enterprises.
Village Industries – They are located in rural area which produces any goods, renders any service with or without the use of power and in which the fixed capital investment per head or artisan or worker is specified by the central government from time to time.
Cottage Industries – They are also known as Rural Industries or Traditional Industries. They are not defined by capital investment criteria but on the basis of characteristics, which are as
follows:
i. These are organised by individuals with private resources.
ii. Normally use family labour and locally available talent.
iii. Simple equipments are used.
iv. Small capital investment.
v. Produce simple products, normally in their own premises.
vi. Production of goods using indigenous technology.
Role of Small Business in Rural India
Small business organizations play an important role in the socio economic development of the country. Some of them are as follows:
a. Multiple sources of income for family.
b. Self employment opportunities in commerce, manufacturing and service segments.
c. Promotion of agro based rural industries.
d. Employment opportunities for artisans and the weaker sections of society.
e. Migration of rural people to urban areas in search of employment has been stopped.
f. Helped to solve the problem of poverty and unemployment.
g. Helped to reduce the income inequalities up to a certain extent.
h. Accelerated industrial growth of the country.
Problems of Small Business – Following are the major problems faced by small business in India:
1. Finance – One of the severe problems faced by SSIs is that of non-availability of adequate finance to carry out its operations.
2. Raw materials – Availability and procurement of raw material is another major problem faced by the SSIs. Their bargaining power is relatively low due to the small quantity of purchases.
3. Managerial skills – SSIs are generally promoted and operated by single person, who may not possess all the managerial skills required to run the business. They are also not in a
position to afford professional managers.
4. Labour – Productivity per employee is relatively low and employee turnover is generally high due to low remuneration. It also faces lack of specialization.
5. Marketing – In most of the cases, marketing is a weaker area of small organisations; therefore exploitation of middlemen is very more.
6. Quality – Many small businesses do not follow the desired standards of quality due to shortage of finance and resources.
7. Capacity utilization – Many of the SSIs are operating below full capacity due to lack of marketing skills or demand. It will cause to increase its operating cost and leads to sickness and closure of the business.
8. Technology – Most of the SSIs use outdated technology, resulting in low productivity and uneconomical production.
9. Sickness – Due to many internal and external problems, most of the SSIs are in the edge of sickness.
10.Global competitions – Most of the SSIs face competitions not only from medium and large industries, but also from Multinational Companies in the areas of quality, technology,
finance, managerial skills etc.
Government Assistance to Small Industries and Small Business Units
Government provides various support measurers and programs for the promotion of small and rural Industries, some of them are given below:
a. National Bank for Agriculture and Rural Development (NABARD)
· Set up in 1982 for integrated rural development.
· Provides finance to small industries, cottage and village industries and artisans.
· Offers counseling and consultancy services.
· Training and development programs for rural entrepreneurs.
b. Rural Small Business Development Centre (RSBDC)
· It is an initiative of world association for small and medium entrepreneurs.
· It is sponsored by NABARD.
· Giving management and technical assistance to micro and small entrepreneurs in rural areas.
· Organizing programs on rural entrepreneurship, skill upgradation workshop,training programs etc.
c. National Small Industries Corporation (NSIC)
· Set up in the year 1955 to promote and foster the growth of SSIs in India.
· Supply of indigenous and imported machines on hire purchase basis.
· Supply of raw materials – locally and imported.
· Support in export of products.
· Monitoring and advisory services.
· Providing latest technology.
· Awareness on technological upgradation.
· Development of software technology parks and technology transfer centres.
d. Rural and Women Entrepreneurship Development (RWED)
· To create a business environment to encourage rural and women entrepreneurs.
· To enhance the productivity of labors and institutions.
· To provide training for women entrepreneurs.
· To give advisory services in all respects.
e. Scheme of Fund for Regeneration of Traditional Industries (SFURTI)
· Established in 2005 to make traditional industries more productive and competitive.
· Implemented by Ministry of Agro and Rural Industries in collaboration with State Governments.
· To develop clusters of traditional industries in various part of the country.
· To make traditional industries more innovative and profitable.
· To create sustainable employment opportunities in traditional sector.
f. District Industries Centres (DICs)
· Established in 1978
· To support small entrepreneurs at district level.
· Provides all facilities and support to set up small and village industries.
· Identification of suitable schemes for entrepreneurs by Central and State Govts.
· Preparation of feasibility reports on each industry.
· Arrangement of credit facilities and equipments.
· Arrangement of raw materials.
· To impart training for artisans, entrepreneurs etc.
Entrepreneurship Development
The word entrepreneur is derived from the French verb entreprende, which means to undertake. Entrepreneurship is the process of setting up of one’s own business. The person
who sets up the business is entrepreneur and the outcome of the process (business unit) is called enterprise.
“Entrepreneur is a person who organizes the business, undertakes the risk and enjoys the profit” – Richard Cantillon_French Economist.
An entrepreneur is basically a businessman and he brings together the factors of production such as land, labour and capital and organizes it. An entrepreneur is more than a businessman,
if a businessman brings some innovation to his activities and eyes on value addition to his products or services, he is called an entrepreneur. In fact, all entrepreneurs are businessmen,
but all businessmen are not entrepreneurs.
“Entrepreneurship is the purposeful activity of an individual or a group of associated individuals, undertaken to initiate, maintain or organize profit oriented business unit for production or distribution of economic goods and services.”
Characteristics of Entrepreneurship
1. Systematic activity – Entrepreneurship is a systematic, step by step and purposeful activity. It requires skill, knowledge and competency which can be acquired, learnt and developed through education, training, observation and experience. Thus we can say the entrepreneurs are made but not born.
2. Lawful and purposeful activity – The aim of entrepreneurship is to run a lawful business.
3. Brings innovation and creativity to the business.
4. Organizes production – The entrepreneur brings the idea of business and the factors of production, thus he organizes the production activities.
5. Risk taking – The entrepreneur takes all the risks in the business as he brings all the factors of production including capital.
Startup India Scheme
Startup India Scheme is an important initiative by Govt. of India to promote a strong ecosystem for nurturing innovation and startup (new enterprises) in the country. As per the notification of the Ministry of Commerce and Industry, a startup means:
1. An entity incorporated or registered in India.
2. Not older than 5 years.
3. Annual turnover does not exceed Rs.25 crores in any preceding year.
4. Working towards innovation, development or comercialisation of products or services with the support technology or Intellectual Property Rights (IPR) and Patents.
Popular Startups in India – Paytm, Flipcart, Snapdeal, Swiggy, Bigbasket, Byju’s App, Ola
Cabs, Make My Trips, ShopClues, OYO Rooms, Zomato, Redbus, Uber Eats etc.
Aims and Objectives of Startup Scheme:
1. Trigger an entrepreneurial culture.
2. Create awareness about the charms of entrepreneurship among the youth.
3. Encourage more dynamic startups by motivating educated youth as a good career.
4. To support the startups in various stages such as pre-startup stage, nascent (beginning stage) and early post startup stage.
5. To promote under represented target groups such as women, back communities, scheduled castes, scheduled tribes etc.
Startup India – Action Point
1. Simplification and hand-holding – Formalities simplified and extended support to the startup ventures.
2. Startup India Hub – To create a single point contact for the entire startup system and to enable knowledge exchange and access to funding.
3. Legal support and fast tracking Patent Examination – To give protection for patents, trademark and designs of innovative startups through SIPP (Startups Intellectual Property Protections).
4. Easy Exist – In the event of failure and wind up of operations, procedures are adopted to reallocate capital and resources towards more productive avenues. Thus the entrepreneurs can easily exit from the business if required.
5. Incubator setup – The government envisages setting up of incubators across the country in PPP mode (Private Public Partnership).
6. Tax exemption – The profit of startup initiatives are exempted from Income Tax for a period of 3 years.
Ways to fund startup
1. Boot Strapping – Self financing by the promoters from their personal savings and resources.
2. Crowd Funding – Pooling resources by a group of people for a common goal especially through internet platforms.
3. Angel Investment – Angel investors are the individuals with surplus cash who have keen interest to invest in startups. They also offer mentoring or advice along with capital.
4. Venture Capital – Venture capitalists provide professionally managed funds to companies and startups that have huge potential. It is also called risk capital as it is invested in new ventures. Eg: Accel Partners, Blume Ventures etc.
5. Business Incubators and Accelerators – Incubators provide funds for startups in the early stage of its business, whereas accelerators help the startups to run or to take a giant leap in business. Eg: Angel Prime, Khosla Labs, Startup Village etc.
6. Microfinance and NBFCs – Microfinance is a category of financial services targeted at individuals and small business who lack access to conventional banking or who have not qualified for a bank loan. Eg: BSS Microfinance P Ltd. , Asirvad Microfinance Pvt. Ltd. etc.
NBFCs (Non Banking Financial Companies) are registered under Indian Companies Act and they perform only lending functions to public and they cannot accept deposits. Eg: Mahindra & Mahindra Financial Services Ltd., Muthoot Finance Ltd., Bajaj Finance Ltd. etc.
Intellectual Property Rights (IPR)
Intellectual Property is a category of property that includes intangible creations of human intellect. The most prominent types of intellectual properties are trade secrets, copyrights, patents, trademarks etc. All inventions begin with an idea. Once the idea becomes an actual product, that idea is treated as an intellectual property. The legal rights conferred on such
products (idea) are called IPR. Once it is allotted to a person by the Govt. authority, he/she can rent, give or sell it to others.
Intellectual property is divided into two categories: Industrial properties like trademarks, industrial designs etc. and copyrights which includes literary and artistic works such as novels, poems, plays, films music, photographs, drawings, paintings, sculptures, architectural designs etc.
Importance of IPR
1. Path-breaking inventions – It encourages new inventions in all segments. Eg: Cancer cure medicines.
2. Incentive – It incentivizes inventors, authors, creators etc. for their work.
3. Helps to prevent loss of income – It allows the inventor to sell the rights to third parties and thus he/she can generate income.
4. Recognition – It helps authors, creators etc. to get recognition for their work.
Famous Legislations and Agreements on IPR
1. TRIPS – Trade-Related Intellectual Property Systems Agreement (a part of WTO).
2. Trade Mark Act 1999.
3. Geographical Indications of Goods (Registration and Protection) Act 1999.
4. Designs Act 2000.
5. Protection of Plant Varieties and Farmers’ Rights Act 2001.
6. Patents Act 2005.
7. Copyright (Amendment) Act 2012.
Types of IPs
1. Copy Right – It is the right to “not copy” conferred upon the creators of literary, artistic, musical, sound recording, films etc.
2. Trademark – Any word, name, or symbol that gives an identity to goods or service made by an individual, company, organization etc. (To register the trademark you can visit www.ipindia.nic.in).
3. Geographical Indication – GI is an identification which identifies agricultural, natural or manufactured products originating from a definite geographical territory. Eg: Banaras Brocades, Kashmiri Pashmina Woolen Shawl, Nagpur Orange etc.
4. Patent – It is an exclusive right granted by the government to prevent others from making, using, offering for sale, selling or importing the invention. For an invention to be patentable, it must be new, non-obvious (not easily discoverable) and having an industrial application.
5. Design – It includes shape, pattern etc. that is applied to any article. Eg: Design of a car, house, bottle etc. The term of protection of a design is valid for 10 years, which can be renewed for further 5 years. After that it will come under public domain.
6. Plant Variety – It is a type of variety which is bred and developed by farmers. Eg: hybrid versions of potatoes, rice, pepper etc. This lead to the growth of seed industry.
7. Semiconductor Integrated Circuits Layout Design – It is used to perform electronic circuitry function. Eg; Computer Chip.
We have provided you Class 11 Small Business and Entrepreneurship Notes Pdf Download with some important Questions and answers and there is a chance that you might observe the same kind of questions here in your exam paper too. Thus, begin learning Small Business and Entrepreneurship Class 11 Important Questions and score great marks.
We trust that you like this Small Business and Entrepreneurship Class 11 notes Pdf and it will help you to make your preparation better. Stay in contact with the page for more related stuff. If you have any questions, you might inform us by writing comments in the below box. | https://cbsencertsolutions.com/small-business-and-entrepreneurship-class-11-business-studies-notes-and-questions/ |
Abstract: Dynamic microscale surface topographies are desired in smart optics, controlling surface wettability and preventing marine biofouling. Voltage-controlled reversible responses have demonstrated potential for reliable reproducibility and stability, fast response, and for actuating thin films fixed over large solid surfaces. To obtain reversible deformation with regular geometric patterns, however, electrical methods have had to be coupled with mechanical stretching/bending and other ways to induce anisotropy. There is a great need and potential for on-demand electrical generation of programmable complex responsive surface patterns. Here we demonstrate a responsive polymer coating over an underlying pattern of counter electrodes which can be activated selectively. We present a patternable electrode printing method to achieve localized and structured wrinkling deformation without mechanical pre-force deformation. We discover that below a minimal separation distance, electrodes below the polymer act as a single electrode. We establish parameters that govern the alignment of wrinkles and quantify the regularity and direction of the new patterns between electrodes separated by larger than this distance. We analyze and quantify the regularity of the formed wrinkling patterns by four disorder metrics: box-counting fractal dimension, tortuosity, angle distribution, and branch number. We demonstrate the application of such electrode/wrinkles-on-demand patterning with a working multi-state light reflection–diffusion–grating device. | https://www.materialstoday.com/amorphous/articles/s136970212030331x/ |
The views expressed in our content reflect individual perspectives and do not represent the official views of the Baha'i Faith.
Friends since their early youth—and most recently sisters-in-law—Haylee Alai and Neda Loh find spiritual comfort, peace, and tranquillity through the art of watercolor.
Their project, titled Water and Wildfire, features their watercolor paintings, all inspired by the sacred writings of the Baha’i Faith, images of the Baha’i shrines and holy places, and the sweet prayers, writings and quotes by the Central Figures of the Baha’i Faith—the Bab, Baha’u’llah, and Abdu’l-Baha.
Despite living on opposite sides of the country, Neda in Broome, a small town located in far North-West Australia, and Haylee in Sydney, they offer their images as budding watercolor painters through a shared account on Instagram under the handle @waterandwildfire. Haylee and Neda use this platform to share their stories and reflections for each post, offering followers insight, context, history, and a personal connection to their work.
In this interview, Neda and Haylee talk about what inspired them to explore the art of watercolor, how sharing gifts can go beyond those who receive them, and how the Baha’i teachings play directly into their shared vision for their art and their lives:
[Shadi] When and why did you start exploring the art of watercolor?
[Neda] I started painting with watercolor on my year of service at the Baha’i World Centre in Haifa, Israel, in 2010. I’ve always enjoyed any type of visual arts and found endless inspiration during my time in the Holy Land. However, having limited access to art stores forced me to get creative and use anything I could find to make art! I had taken one stick of watercolor paints with me and they became my sole medium for the 18 months I was volunteering there! I even used to grind up old make up, coffee and tea to find new colors and textures. I haven’t been able to experiment like that since!
[Haylee] My desire to paint is linked very heavily to why I use the Baha'i writings in my art. I started watercoloring because I enjoyed drawing and painting so much in the Ruhi study groups I was a part of, and realized that it was a great way to meditate and reflect on the beautiful writings and concepts of the Baha’i Faith. My real interest in watercolor actually developed in 2012 when I saw some of Neda's posts on Facebook that included images of her paintings. The way that she integrated watercolor, the Baha'i holy places and the Baha'i writings touched my heart immediately. As I developed my skill, I began creating watercolor pieces with quotations from the Baha'i writings as gifts for friends, and the act of giving these personalized gifts filled me with joy and I could tell it meant a lot to those who received them.
[Shadi] What inspired your collaboration?
[Neda] I remember the first time I saw one of Haylee’s watercolor paintings, I thought she had been painting for years and years! Her dedication to learning a skill and excelling in it is so inspiring and she’s played a big part in giving me motivation to paint and create artworks. Her creative striving reminds me of this quote by Abdu’l-Baha, where he encourages us to work hard and excel in our craft:
I rejoice to hear that thou takest pains with thine art, for in this wonderful new age, art is worship. The more thou strivest to perfect it, the closer wilt thou come to God. What bestowal could be greater than this, that one's art should be even as the act of worshipping the Lord? That is to say, when thy fingers grasp the paintbrush, it is as if thou wert at prayer in the Temple. - Abdu'l-Baha, from a Persian tablet.
Needless to say, having her as a sister-in-law has been a huge bonus! Haylee also organized a few painting sessions with other youth, which allowed us to spend time together in a creative context. It’s different to catching up over a meal, somehow there is less pressure to make small talk and easier to speak about more elevated concepts. I have tried to carry this where I live and serve and it’s been well-received by young people in particular!
[Haylee] I would say that we have been collaborating "in life" for many years now, and that has now recently evolved into sharing this Instagram account together. I feel that being able to serve the community and participate in Baha'i study courses and seminars together has united our vision and desire for similar things in life. When we both lived in Perth we had the opportunity to paint together and express our constant admiration for each other! I am lucky enough to have Neda as my sister-in-law now, and it just felt natural that we should start this project together.
[Shadi] What inspired you to use Baha'i prayers and writings in your work?
[Neda] I often wonder about why the Baha’i writings are referred to as the “creative word.” I think the soul-stirring nature of the word of God must spark the imagination of so many, and will be the source of countless inventions and artistic creations. As an artist, finding inspiration can be difficult, but the Baha'i teachings never fail in that sense. When in London, Abdu’l-Baha was reported to have said this regarding the influence the holy writings and the highest purpose of art:
When the Light of the Sun of Truth inspires the mind of a painter, he produces marvellous pictures. These gifts are fulfilling their highest purpose, when showing forth the praise of God. – Abdu’l-Baha, quoted by Lady Blomfield in The Chosen Highway, p. 167.
[Haylee] As I mentioned before, it was my participation in a series of Baha'i courses called the Ruhi Institute that prompted my use of the Baha'i writings in my art. I use the Baha'i writings because the concepts, poetry and power of the words are so profound, and I am always able to find a Baha’i quote that seems to suit every situation and make each gift filled with so much love.
[Shadi] What inspires your images?
[Neda] The natural world, the simplicity and beauty of the architecture at the Baha’i World Centre buildings and the Baha’i Houses of Worship—anything and any space that reminds me of our spiritual nature.
[Shadi] How do you choose your specific quotes and associated images?
[Haylee] As most of my paintings have always been given as gifts to friends, I would say that this has had a big impact on the inspiration for the images and quotes. When I know that there is an occasion where I can create a gift for someone, I spend a few days thinking to myself and being open to inspiration from the writings and from nature. At some point, the right quote and image that would match it become clear and then I can begin painting. Sometimes I might think of the Baha'i quote first, and then have to think more about what image would suit it, and sometimes I have an image in my mind and then it is a few days until the right quote becomes clear. To me, the process of thinking of a friend, and choosing a quotation and image that would mean something to them and touch their heart is a really sincere way of showing how much you care about someone and are thinking of their wellbeing.
For example, I created a piece for my friends wedding, which included a quote by Baha’u’llah:
Make me ready, in all circumstances, O my Lord, to serve Thee and to set myself towards the adored sanctuary of Thy Revelation and of Thy Beauty. If it be Thy pleasure, make me to grow as a tender herb in the meadows of Thy grace, that the gentle winds of Thy will may stir me up and bend me into conformity with Thy pleasure, in such wise that my movement and my stillness may be wholly directed by Thee. - Prayers and Meditations, p. 240.
[Shadi] What makes Water and Wildfire unique?
[Haylee] Honestly I don't feel like my work stands out, and I don't feel that it is unique at all! The Baha'i writings are there for everyone to use and be inspired by. The painting that I do is painting that anyone could learn over time (watching Instagram videos of your favorite artists can really help!). The way that I view Water and Wildfire is that it is a way of encouraging others to create art and share it with the world. Beauty takes so many forms, but I believe that an important part of creating beautiful art is the intentions and sincerity with which we do it. Taking time out of our day to engage in the arts is good for our soul and helps us to reflect on life and what is important! It is helpful to remind ourselves of what Baha’u’llah has told us about the role of the arts: “Arts, crafts and sciences uplift the world of being, and are conducive to its exaltation.” - Epistle to the Son of the Wolf, p. 26.
[Shadi] If readers want to learn more about your work and get in touch where should they go? | https://bahaiteachings.org/water-wildfire-two-sisters-paint-bahai-writings/ |
These well researched mindfulness-based approaches incorporate mindfulness training that can lead to clinically significant improvements in psychological functioning to individuals with a wide range of conditions.
Groups are conducted periodically and are subject to change.
BEING MINDFUL: is an introductory program combining education about mindfulness concepts with experiential mindfulness skills training and sensory awareness practices.
MINDFULNESS APPROACH TO ART THERAPY: Guided to the basic principles of Mindfulness, this is an art therapy group centred on living life in the way that you value most. By using art making materials in supportive group setting, members are encouraged to experience self-reflection and self-understanding.
MINDFULNESS BASED COGNITIVE THERAPY (MBCT): is an approach incorporating mindfulness practices with traditional change strategies of CBT and is a way of alleviating depression and addressing recurring episodes.
MINDFULNESS BASED STRESS REDUCTION (MBSR): is a systematic person-centred educational approach that uses intensive mindfulness meditation as the focus to alleviate stress, panic, depression, chronic pain and a range of medical conditions.
MINDFULNESS FOR DAILY LIFE: Explores a greater understanding of self, our actions, our urges, impulses and increases healthy responses to feelings.
MINDFULNESS FOR EMOTIONAL WELLBEING: Improve your understanding of emotions and unhelpful thinking styles through mindfulness practice. Skills, strategies and mindfulness practice will be offered to assist individuals to gain experience and understanding of their emotions, increase self-compassion, develop helpful techniques and identify unhelpful thinking styles to minimise them. | http://delmodev.aus.net/services/day-programs/mindfulness-based-groups |
The relationship between two variables can be represented graphically.
When the rate of change for both variables is constant, the relationship between these two variables is represented by a straight line.
When the rate of change is constantly varying, the relationship between the two variables is represented graphically by a curve.
Instantaneous rate of change is the rate of change at any particular point on the curve.
The following are some examples of instantaneous rate of change in real life.
1. Velocity
Velocity is used to describe the rate of change of position by an object in respect to time.
Velocity describes how objects are moving through space, over the ground or in the air i.e. their displacement.
It describes how fast the object is moving at anywhere along a given path.
Car manufacturers use instantaneous velocity to describe how powerful their engines are. For example, a manufacturer may say that their car can go from zero to 100mph in 8 seconds.
The velocity of the car at a given time (8 seconds) will be 100mph.
2. Acceleration
Acceleration describes the rate of change in velocity of an object with respect to time. It describes the change in velocity of a moving object at a given instant in time.
For example, acceleration can be used in sports such as baseball in determining how effective a training regimen is.
Team statisticians will consider the acceleration of the baseball as it is struck by a bat to determine improvements in power of players.
3. Population growth rate
Population growth rate is used to analyze the rate at which a population increases.
It can be used to predict the population of a specified region at a specific time.
Biologists use population growth rate to determine by how much a colony of bacteria will have grown in a given period of time and therefore identify conditions that encourage or discourage their growth.
4. Mortality rate
Mortality rate is used to describe the number of deaths that occur in a given population within a specified period.
Epidemiologists use mortality rate to predict the rate of deaths within a specified period.
Mortality rate helps public health authorities better gauge the performance of their health care system.
It is also used to determine the impact of threats to health such as nutritional deficiencies or infectious diseases.
5. Marginal revenue
Marginal revenue is used by businesses in cost-benefit analysis. It is the increase in revenue that a business would earn from adding a single unit to their output.
Businesses use marginal revenue to determine by how much they can increase their income if they were to add a single unit to their sales before their revenue will begin to slow down.
Companies use this model to maximize their profits. They can determine the optimum levels of production or stocking in products to produce maximum profits.
Firms also use this to determine whether they should halt the production of an item or the offering of a service.
6. Instantaneous rate of a reaction
The instantaneous rate of a reaction is the rate at which a chemical reaction is taking place at a given moment in time.
It is a measure of the change in concentration of the reactants and the products at any specific time.
Test strips used by healthcare workers apply this concept. Health workers use disposable test strips to determine the concentration of various substances in bodily fluids such as urine.
The test strips are dipped in the urine sample. The healthcare worker will wait for a specified amount of time to get a reading.
The results shown on the test strips are a reflection the concentration of reactants and products at a given moment in time.
7. Instantaneous voltage
Instantaneous voltage is the value of alternating current voltage in a cycle at a particular instant in time. It is the voltage between two given points at any given moment in time.
Electricians use instantaneous voltage to determine the desired voltage across different parts of a circuit. They use this to determine what capacitors and inductors to include in printed circuit board(PCB) design. | https://boffinsportal.com/7-instantaneous-rate-of-change-real-life-examples/ |
Accessible telecommunication technology is increasingly important for people who are working remotely and relying on real-time communication (RTC) for daily needs.
Web Real-time communications (WebRTC) is a series of protocols that support quick direct browser-to-browser transfer of call (and other data) without the need for a brittle plugin architecture. This transfer is nearly instantaneous with very small degrees of latency, and is considered a robust way of supporting peer-to-peer based communication. WebRTC technologies enable better video/audio calls, instant messaging text chat, file exchange, internet relay chat, screen sharing and more.
To make this happen the Internet Engineering Task Force (IETF) and W3C have collaborated since 2011 to standardize the necessary protocols and APIs, in particular the core WebRTC 1.0: Real-time Communication Between Browsers API specification. This spec defines a set of ECMAScript APIs that allow media to be sent to and received from another browser or device using an appropriate set of real-time protocols.
So what does this mean for the people with disabilities? How can people with disabilities effectively take advantage of the potential benefits that these real-time communication (RTC) protocols allow? What are the challenges for people with disabilities using RTC applications?
Challenges for People with disabilities using RTC applications
For real-time communication, video conferencing and instant messaging some of the challenges faced are; the need to route audio output to different sound cards or output devices, to support captioning and live transcription, and the transfer of alternate formats such as sign-language. The user may also need to be able to perform status polling (where on a call a user can query who is talking, or muted, on the call etc in a way that works with assistive technologies).
There are also different flavors of messaging preferred by blind users and Real-time text (RTT) needs for deaf and hard of hearing users.
Federal Communications Commission (FCC) and European requirements for Real-time text (RTT)
On December 15, 2016, the FCC adopted rules to facilitate a transition from text telephony (TTY) technology to Real-time text (RTT) technology.
TTY has been used by deaf and hard of hearing people to communicate over phone lines by text. TTY usage is declining. Now Real-time text allows the deaf and hard of hearing community to engage in more conversational style interactions that are critical for emergency services such as 911 and relay services such as 711. Real-time text has other advantages and is requirement in the European procurement standard EN 301 549 (PDF). It is therefore important that RTC applications support Real-time text.
RTC Accessibility User Requirements (RAUR) – First Public Working Draft published
Real-time communication applications have the potential to address these challenges but firstly designers and developers have to understand accessibility user needs. W3C/WAI is active in this space and in order to layout these challenges the Research Questions Task Force (RQTF) have published ‘RTC Accessibility User Requirements (RAUR)’ as a first public working draft.
The RQTF is a task force of the Accessible Platform Architectures (APA) working group at W3C. The APA works to ensure W3C specifications provide support for accessibility to people with disabilities.
RTC Accessibility User Requirements (RAUR) is designed to inform the reader of potential user needs for people with disabilities. It will also help initiate discussion and gather feedback on the best ways to address current gaps. The term RTC is used throughout to denote a range of technologies (including WebRTC) that when combined help to create accessible RTC applications.
From these user needs were drafted a set of user requirements that may be implemented at a system or platform level. Some may be authoring requirements.
Please note, in the context of this document we are outlining generic draft user requirements only. Those listed are not normative. This means they do not represent a series of things that you must do to conform to any current accessibility standard. This is a first public draft of what may go on to inform the development of other accessibility guidelines.
We welcome your feedback!
Your feedback will help inform our future work in both accessibility guidelines and technology specifications. We are therefore actively looking for your input on the development of these user needs and their related requirements. | https://www.w3.org/blog/2020/03/rtc-accessibility-user-requirements-call-for-review/ |
This application is a National Stage application of International Application No. PCT/IB2018/059413, filed Nov. 28, 2018, which claims priority to U.S. Patent Application No. 62/592,037, filed on Nov. 29, 2017.
The Sequence Listing, which is a part of the present disclosure, is submitted concurrently with the specification as a text file. The name of the text file containing the Sequence Listing is “1712288A_Seqlisting.txt”, which was created on Apr. 27, 2020 and is 1,624,362 bytes in size. The subject matter of the Sequence Listing is incorporated herein in its entirety by reference.
INCORPORATION BY REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY
FIELD OF THE INVENTION
The present invention relates in general to methods for conferring on plants agricultural level tolerance to herbicides. Particularly, the invention refers to plants having an increased tolerance to PPO-inhibiting herbicides. More specifically, the present invention relates to methods and plants obtained by mutagenesis and cross-breeding and transformation that have an increased tolerance to PPO-inhibiting herbicides.
BACKGROUND OF THE INVENTION
Herbicides that inhibit protoporphyrinogen oxidase (hereinafter referred to as Protox or PPO; EC:1.3.3.4), a key enzyme in the biosynthesis of protoporphyrin IX, have been used for selective weed control since the 1960s. PPO catalyzes the last common step in chlorophyll and heme biosynthesis which is the oxidation of protoporphyrinogen IX to protoporphyrin IX. (Matringe et al. 1989. Biochem. 1. 260: 231). PPO-inhibiting herbicides include many different structural classes of molecules (Duke et al. 1991. Weed Sci. 39: 465; Nandihalli et al. 1992. Pesticide Biochem. Physiol. 43: 193; Matringe et al. 1989. FEBS Lett. 245: 35; Yanase and Andoh. 1989. Pesticide Biochem. Physiol. 35: 70). These herbicidal compounds include the diphenylethers {e.g. lactofen, (+−)-2-ethoxy-1-methyl-2-oxoethyl 5-{2-chloro-4-(trifluoromethyl)phenoxy}-2-nitrobenzoate; acifluorfen, 5-{2-chloro-4-(trifluoromethyl)phenoxy}-2-nitrobenzoic acid; its methyl ester; or oxyfluorfen, 2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluorobenzene)}, oxidiazoles, (e.g. oxidiazon, 3-{2,4-dichloro-5-(1-methylethoxy)phenyl}-5-(1,1-dimethylethyl)-1,3,4-oxadiazol-2-(3H)-one), cyclic imides (e.g. S-23142, N-(4-chloro-2-fluoro-5-propargyloxyphenyl)-3,4,5,6-tetrahydrophthalimide; chlorophthalim, N-(4-chlorophenyl)-3,4,5,6-tetrahydrophthalimide), phenyl pyrazoles (e.g. TNPP-ethyl, ethyl 2-{1-(2,3,4-trichlorophenyl)-4-nitropyrazolyl-5-oxy}propionate; M&B 39279), pyridine derivatives (e.g. LS 82-556), and phenopylate and its O-phenylpyrrolidino- and piperidinocarbamate analogs. Many of these compounds competitively inhibit the normal reaction catalyzed by the enzyme, apparently acting as substrate analogs.
Application of PPO-inhibiting herbicides results in the accumulation of protoporphyrinogen IX in the chloroplast and mitochondria, which is believed to leak into the cytosol where it is oxidized by a peroxidase. When exposed to light, protoporphyrin IX causes formation of singlet oxygen in the cytosol and the formation of other reactive oxygen species, which can cause lipid peroxidation and membrane disruption leading to rapid cell death (Lee et al. 1993. Plant Physiol. 102: 881).
Escherichia coli
Bacillus subtilis
Chlamydomonas reinhardtii
E. coli
Not all PPO enzymes are sensitive to herbicides which inhibit plant PPO enzymes. Both the and PPO enzymes (Sasarmen et al. 1993. Can. J. Microbiol. 39: 1155; Dailey et al. 1994. J. Biol. Chem. 269: 813) are resistant to these herbicidal inhibitors. Mutants of the unicellular alga resistant to the phenylimide herbicide S-23142 have been reported (Kataoka et al. 1990. J. Pesticide Sci. 15: 449; Shibata et al. 1992. In Research in Photosynthesis, Vol. III, N. Murata, ed. Kluwer:Netherlands. pp. 567-70). At least one of these mutants appears to have an altered PPO activity that is resistant not only to the herbicidal inhibitor on which the mutant was selected, but also to other classes of protox inhibitors (Oshio et al. 1993. Z. Naturforsch. 48c: 339; Sato et al. 1994. In ACS Symposium on Porphyric Pesticides, S. Duke, ed. ACS Press: Washington, D.C.). A mutant tobacco cell line has also been reported that is resistant to the inhibitor S-21432 (Che et al. 1993. Z. Naturforsch. 48c: 350). Auxotrophic mutants have been used to confirm the herbicide resistance of cloned plant PPO-inhibiting herbicides.
Three main strategies are available for making plants tolerant to herbicides, i.e. (1) detoxifying the herbicide with an enzyme which transforms the herbicide, or its active metabolite, into non-toxic products, such as, for example, the enzymes for tolerance to bromoxynil or to basta (EP242236, EP337899); (2) mutating the target enzyme into a functional enzyme which is less sensitive to the herbicide, or to its active metabolite, such as, for example, the enzymes for tolerance to glyphosate (EP293356, Padgette S. R. et al., J. Biol. Chem., 266, 33, 1991); or (3) overexpressing the sensitive enzyme so as to produce quantities of the target enzyme in the plant which are sufficient in relation to the herbicide, in view of the kinetic constants of this enzyme, so as to have enough of the functional enzyme available despite the presence of its inhibitor. The third strategy was described for successfully obtaining plants which were tolerant to PPO inhibitors (see e.g. U.S. Pat. No. 5,767,373 or 5,939,602, and patent family members thereof.). In addition, US 2010/0100988 and WO 2007/024739 discloses nucleotide sequences encoding amino acid sequences having enzymatic activity such that the amino acid sequences are resistant to PPO inhibitor herbicidal chemicals, in particular 3-phenyluracil inhibitor specific PPO mutants.
Amaranthus
Amaranthus
Amaranthus
Amaranthus
Amaranthus
Amaranthus
Alopecurus myosuroides
WO 2012/080975 discloses plants the tolerance of which to a PPO-inhibiting herbicide named (1,5-dimethyl-6-thioxo-3-(2,2,7-trifluoro-3-oxo-4-(prop-2-ynyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-6-yl)-1,3,5-triazinane-2,4-dione) had been increased by transforming said plants with nucleic acids encoding mutated PPO mutated enzymes. In particular, WO 2012/080975 discloses that the introduction of nucleic acids which code for a mutated PPO of an type II PPO in which the Arginine at position 128 had been replaced by a leucine, alanine, or valine, and the phenylalanine at position 420 had been replaced by a methionine, cysteine, isoleucine, leucine, or threonine, confers increased tolerance/resistance to a benzoxazinone-derivative herbicide. WO 2013/189984 discloses plants the tolerance of which to PPO inhibitors had been increased by transforming said plants with nucleic acids encoding mutated PPO enzymes, in which the Leucine corresponding to position 397 of an type II PPO had been replaced, and the phenylalanine corresponding to position 420 of an type II PPO had been replaced. WO2015/022636 discloses plants the tolerance of which to PPO inhibitors had been increased by transforming said plants with nucleic acids encoding mutated PPO enzymes, in which the Arginine corresponding to position 128 of an type II PPO had been replaced, and the phenylalanine corresponding to position 420 of an type II PPO had been replaced, but the replacement occurred with amino acids, which are different from those disclosed in WO 2012/080975. WO2015/092706 describes PPO polypeptides from a plurality of organisms, which PPO polypeptides had been mutated to comprise the advantageous mutations employed for the type II PPO. WO2015/022640 discloses PPO polypeptides from and mutants thereof, which confer tolerance to a broad spectrum of PPO inhibiting herbicides.
The inventors of the present invention have now surprisingly found that those types of mutants confer increased tolerance/resistance to a new class of PPO inhibitors, hereinafter described as uracilpyridines or uracilpyridine herbicides. Thus, to date, the prior art has not described uracilpyridine tolerant plants containing a mutated PPO nucleic acid according to the present invention, which are tolerant/resistant to a broad selection of uracilpyridines.
Therefore, what is needed in the art are crop plants and crop plants having increased tolerance to uracilpyridines and containing at least one wildtype and/or mutated PPO nucleic acid according to the present invention. Also needed are methods for controlling weed growth in the vicinity of such crop plants or crop plants. These compositions and methods would allow for the use of spray over techniques when applying herbicides to areas containing crop plants or crop plants.
SUMMARY OF THE INVENTION
a) providing, at said site, a plant that comprises at least one nucleic acid comprising a nucleotide sequence encoding a protoporphyrinogen oxidase (PPO) polypeptide which is resistant or tolerant to a “PPO inhibiting herbicide”;
b) applying to said site an effective amount of said herbicide, wherein the PPO inhibiting herbicide is a uracilpyridine of formula (I)
The problem is solved by the present invention which refers to a method for controlling undesired vegetation at a plant cultivation site, the method comprising the steps of:
wherein the substituents have the following meanings:
1
2
1
6
3
6
Rhydrogen, NH, C-C-alkyl or C-C-alkynyl;
2
1
6
1
6
Rhydrogen, C-C-alkyl or C-C-haloalkyl;
3
1
6
Rhydrogen or C-C-alkyl;
4
RH or halogen;
5
2
2
3
2
Rhalogen, CN, NO, NH, CFor C(═S)NH;
6
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
RH, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy, C-C-alkylthio, (C-C-alkyl)amino, di(C-C-alkyl)amino, C-C-alkoxy-C-C-alkyl, C-C-alkoxycarbonyl;
7
1
3
1
3
RH, halogen, C-C-alkyl, C-C-alkoxy;
8
9
9
10
11
9
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
—N═CRR, wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
3
6
3
6
1
6
3
6
3
6
1
6
1
4
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, C-C-heterocyclyl-C-C-alkyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
11
9
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
R, Rindependently of one another are R, or together form a 3- to 7-membered carbocyclus,
ROR, SR, NRR, NROR, NRS(O)Ror NRS(O)NRR, wherein
n 1 to 3;
2
2
1
3
Q CH, O, S, SO, SO, NH or (C-C-alkyl)N;
W O or S;
3
X NH, NCH, O or S;
Y O or S;
1
6
1
6
1
6
1
6
Z phenyl, pyridyl, pyridazinyl, pyrimidinyl or pyrazinyl, each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
including their agriculturally acceptable salts or derivatives, provided the compounds of formula (I) have a carboxyl group
and wherein the effective amount of said herbicide does not kill or inhibit the growth of the herbicide-tolerant plant of a).
a. Motif 1: SQ[N/K/H]KRYI, wherein the Arg at position 5 within said motif is substituted by any other amino acid;
b. Motif 2: TLGTLFSS, wherein the Leu at position 2, and/or the Gly at position 3, and/or the Leu at position 5 within said motif is substituted by any other amino acid;
c. Motif 3: [F/Y]TTF[V/I]GG, wherein the Phe at position 4 within said motif is substituted by any other amino acid.
In one embodiment, the herbicide resistant or tolerant PPO polypeptide comprises one or more of the following motifs 1, 2, and/or 3:
a. the amino acid corresponding to Arg128 of SEQ ID NO: 1 is substituted by any other amino acid.
b. The amino acid corresponding to Gly211 of SEQ ID NO: 1 is substituted by any other amino acid
c. the amino acid corresponding to Leu397 of SEQ ID NO: 1 is substituted by any other amino acid.
d. the amino acid corresponding to Gly398 of SEQ ID NO: 1 is substituted by any other amino acid
e. the amino acid corresponding to Leu400 of SEQ ID NO: 1 is substituted by any other amino acid
f. the amino acid corresponding to Phe420 of SEQ ID NO: 1 is substituted by any other amino acid.
In another embodiment, the herbicide resistant or tolerant PPO polypeptide comprises a variant of the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, which variant comprises one or more of the following substitutions:
In still another embodiment, the herbicide resistant or tolerant PPO polypeptide comprises the amino acid sequence of SEQ ID NO: 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, or a variant thereof.
In still another embodiment, the herbicide resistant or tolerant PPO polypeptide comprises the amino acid sequence of SEQ ID NO: 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262,or 264.
a) generating a library of mutated PPO-encoding nucleic acids,
b) screening a population of the resulting mutated PPO-encoding nucleic acids by expressing each of said nucleic acids in a cell or plant and treating said cell or plant with a uracilpyridine,
c) comparing the uracilpyridine herbicide-tolerance levels provided by said population of mutated PPO encoding nucleic acids with the uracilpyridine-tolerance level provided by a control PPO-encoding nucleic acid,
d) selecting at least one mutated PPO-encoding nucleic acid that provides a significantly increased level of tolerance to a uracilpyridine as compared to that provided by the control PPO-encoding nucleic acid.
Another object refers to a method of identifying a nucleotide sequence encoding a mutated PPO which is resistant or tolerant to a uracilpyridine herbicide, the method comprising:
In a preferred embodiment, the mutated PPO-encoding nucleic acid selected in step d) provides at least 2-fold as much tolerance to a uracilpyridine as compared to that provided by the control PPO-encoding nucleic acid.
The resistance or tolerance can be determined by generating a transgenic plant comprising a nucleic acid sequence of the library of step a) and comparing said transgenic plant with a control plant.
a) identifying an effective amount of a uracilpyridine in a culture of plant cells or green algae.
b) treating said plant cells or green algae with a mutagenizing agent,
c) contacting said mutagenized cells population with an effective amount of uracilpyridine herbicide, identified in a),
d) selecting at least one cell surviving these test conditions,
e) PCR-amplification and sequencing of PPO genes from cells selected in d) and comparing such sequences to wild-type PPO gene sequences, respectively.
Another object refers to a method of identifying a plant or algae containing a nucleic acid encoding a mutated PPO which is resistant or tolerant to a uracilpyridine, the method comprising:
In a preferred embodiment, the mutagenizing agent is ethylmethanesulfonate.
In another embodiment, the invention refers to a plant cell transformed by and expressing a wild-type or a mutated PPO nucleic acid according to the present invention or a plant which has been mutated to obtain a plant expressing, preferably over-expressing a wild-type or a mutated PPO nucleic acid according to the present invention, wherein expression of said nucleic acid in the plant cell results in increased resistance or tolerance to a uracilpyridine as compared to a wild type variety of the plant cell.
In another embodiment, the invention refers to a plant that expresses a mutagenized or recombinant mutated PPO polypeptide, and wherein said mutated PPO confers upon the plant increased uracilpyridine tolerance as compared to the corresponding wild-type variety of the plant when expressed therein.
In another embodiment, the invention refers to a plant comprising a plant cell according to the present invention, wherein expression of the nucleic acid in the plant results in the plant's increased resistance to uracilpyridine herbicide as compared to a wild type variety of the plant.
The plants of the present invention can be transgenic or non-transgenic.
Preferably, the expression of the nucleic acid of the invention in the plant results in the plant's increased resistance to uracilpyridine herbicides as compared to a wild type variety of the plant.
a) growing said plant; and
b) applying a herbicide composition comprising a uracilpyridine herbicide to the plant and weeds, wherein the herbicide normally inhibits protoporphyrinogen oxidase, at a level of the herbicide that would inhibit the growth of a corresponding wild-type plant.
In another embodiment, the invention refers to a method for growing the plant according to the present invention while controlling weeds in the vicinity of said plant, said method comprising the steps of:
In another embodiment, the invention refers to a seed produced by a transgenic plant comprising a plant cell of the present invention, or to a seed produced by the non-transgenic plant that expresses a mutagenized PPO polypeptide, wherein the seed is true breeding for an increased resistance to a uracilpyridine herbicide as compared to a wild type variety of the seed.
In another embodiment, the invention refers to a method of producing a transgenic plant cell with an increased resistance to a uracilpyridine herbicide as compared to a wild type variety of the plant cell comprising, transforming the plant cell with an expression cassette comprising a wild-type or a mutated PPO nucleic acid.
In another embodiment, the invention refers to a method of producing a transgenic plant comprising, (a) transforming a plant cell with an expression cassette comprising a wild-type or a mutated PPO nucleic acid, and (b) generating a plant with an increased resistance to uracilpyridine herbicide from the plant cell.
Preferably, the expression cassette further comprises a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant.
In another embodiment, the invention relates to using the mutated PPO of the invention as selectable marker. The invention provides a method of identifying or selecting a transformed plant cell, plant tissue, plant or part thereof comprising a) providing a transformed plant cell, plant tissue, plant or part thereof, wherein said transformed plant cell, plant tissue, plant or part thereof comprises an isolated nucleic acid encoding a mutated PPO polypeptide of the invention as described hereinafter, wherein the polypeptide is used as a selection marker, and wherein said transformed plant cell, plant tissue, plant or part thereof may optionally comprise a further isolated nucleic acid of interest; b) contacting the transformed plant cell, plant tissue, plant or part thereof with at least one PPO-inhibiting inhibiting compound; c) determining whether the plant cell, plant tissue, plant or part thereof is affected by the inhibitor or inhibiting compound; and d) identifying or selecting the transformed plant cell, plant tissue, plant or part thereof.
The invention is also embodied in purified mutated PPO proteins that contain the mutations described herein, which are useful in molecular modeling studies to design further improvements to herbicide tolerance. Methods of protein purification are well known, and can be readily accomplished using commercially available products or specially designed methods, as set forth for example, in Protein Biotechnology, Walsh and Headon (Wiley, 1994).
In another embodiment, the invention relates to a combination useful for weed control, comprising (a) a polynucleotide encoding a mutated PPO polypeptide according to the present invention, which polynucleotide is capable of being expressed in a plant to thereby provide to that plant tolerance to a uracilpyridine herbicide; and (b) a uracilpyridine herbicide.
In another embodiment, the invention relates to a process for preparing a combination useful for weed control comprising (a) providing a polynucleotide encoding a mutated PPO polypeptide according to the present invention, which polynucleotide is capable of being expressed in a plant to thereby provide to that plant tolerance to a uracilpyridine herbicide; and (b) providing a uracilpyridine herbicide.
In a preferred embodiment, said step of providing a polynucleotide comprises providing a plant containing the polynucleotide.
In another preferred embodiment, said step of providing a polynucleotide comprises providing a seed containing the polynucleotide.
In another preferred embodiment, said process further comprises a step of applying the uracilpyridine herbicide to the seed.
In another embodiment, the invention relates to the use of a combination useful for weed control, comprising (a) a polynucleotide encoding a mutated PPO polypeptide according to the present invention, which polynucleotide is capable of being expressed in a plant to thereby provide to that plant tolerance to a uracilpyridine herbicide; and (b) a uracilpyridine herbicide, to control weeds at a plant cultivation site.
BRIEF DESCRIPTION OF THE DRAWINGS
(NB: full name of used uracilpyridines is given in the Example section)
FIG. 1
: Rice plants 7 days after treatment with 3.125 g AI/ha of Uracilpyridine 4. A) wild-type, B) R139L mutant, C) R139G mutant, D) G225R mutant, E) G420A mutant, F) F442L mutant, G) F442I mutant, H) F442V mutant, I) L419 mutant, J) L422F mutant
FIG. 2
: SRice plants 7 days after treatment with 6.25 g AI/ha of Uracilpyridine 2. A) wild-type, B) R139L mutant, C) R139G mutant, D) G225R mutant, E) G420A mutant, F) F442L mutant, G) F442I mutant, H) F442V mutant, 1) L419 mutant, J) L422F mutant.
FIG. 3
: Response of rice containing mutations in the OsPPO2 gene and wild-type rice to application of Uracilpyridine 4. Each bar represents the average of up to four plants. Phytotoxicity was assessed 7 days after herbicide application. ***P-value<0.001, **P-value<0.01, *P-value<0.05 as calculated by ANOVA using a QuasiPoisson distribution
FIG. 4
: Response of rice containing mutations in the OsPPO2 gene and wild-type rice to application of Uracilpyridine 2. Each bar represents the average of up to four plants. Phytotoxicity was assessed 7 days after herbicide application. ***P-value<0.001, **P-value<0.01, *P-value<0.05 as calculated by ANOVA using a QuasiPoisson distribution.
Arabidopsis
Arabidopsis
Figure: Transgenic T2 plants harboring PPO tolerance trait, sprayed post in the greenhouse with the following amounts (80, 30, 10, 5, 1 g/ha from right to left) of Uracilpyridine+1% (v/v) MSO. Pictures taken 14 Days After Treatment (DAT). A) is tested with Uracilpyridine 4, B) is tested with Uracilpyridine 5, C) is tested with Uracilpyridine 2, D) is tested with Uracilpyridine 3.
Arabidopsis
1=non-transformed plants
Arabidopsis
2=Transformants with PPO1 wildtype
Arabidopsis
3=Transformants with PPO1 double mutant S305L_Y426M
FIG. 6
Arabidopsis
: Transgenic T2 plants harboring various PPO tolerance trait, sprayed post in the greenhouse with the following amounts (200, 100, 50 g/ha) of Uracilpyridine+1% (v/v) MSO. Pictures taken 14 Days After Treatment (DAT). A) is tested with Uracilpyridine 2, B) is tested with Uracilpyridine 4, C) is tested with Uracilpyridine 1, D) is tested with Uracilpyridine 2.
Arabidopsis
1=non-transformed plants
2=transformed with AMATU_PPO2_TPL_hemG (event R) (SEQ ID NO: 246)
3=transformed with AMATU_PPO2_TPL_hemG (event Q) (SEQ ID NO: 246)
4=transformed with ALOMY_PPO2_TP_hemG (SEQ ID NO: 248)
5=transformed with ALOMY_PPO2_R137L_F438V
6=transformed with ALOMY_PPO2_TPL_AMATU_PPO2_R128M_F420I (SEQ ID NO: 258)
7=transformed with AMATU_PPO2_R128M_F420 I
8=transformed with AMATU_PPO2_R128A_F420M
FIG. 7
: Transgenic Corn plants harboring a PPO inhibitor tolerance trait, sprayed post at the V5 leaf stage with 100 g/ha of Uracilpyridine 2+1% (v/v) MSO. Evaluation performed 14 Days After Treatment (DAT) and is shown as injury (%) relative to non-transgenic treated plants. Application performed on corn in V5 leaf stage with 100 g/ha Uracilpyridine 2. Picture taken at 2 DAT
1=Corn transformed with AMATU_PPO2_R128A_F420L
2=Corn transformed with AMATU_PPO2_R128A_F420
3=non-transformed corn plants
FIG. 8
: Transgenic T3 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed post at the V3 leaf stage with 100 g/ha of Uracilpyridine 2+1% (v/v) MSO. Evaluation performed 14 Days After Treatment (DAT) and is shown as injury (%) relative to non-transgenic treated plants. Application performed on soy at V3 leaf stage with 100 g/ha uracilpyridine 2. Picture taken at 2 DAT
1=Soy transformed with AMATU_PPO2_R128A, F420I
2=Soy transformed with AMATU_PPO2_L397E, F420V
3=Soy transformed with AMATU_PPO2_L397E, F420M
4=Soy transformed with AMATU_PPO2_L397Q, F420M
5=non-transformed soy plants
FIG. 9
: Transgenic T4 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed with 25 g/ha (Panels A-B), 50 g/ha (Panels C-D), 100 g/ha (Panels E-F) of Uracilpyridine 10+1% (v/v) MSO. Evaluation and pictures taken 7 Days After Treatment. Application performed on soybean 14 days after sowing. Panels A, C, & E each show plants harboring a PPO trait containing R128A+F420L, R128A+F420I AMTU PPO2 variants as well as untransformed soybean (last column). Panels B, D, & F each show plants harboring a PPO trait containing L397Q+F420V and L397E+F420M AMTU PPO2 variants.
FIG. 10
: Transgenic T4 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed with 25 g/ha (Panels A-B), 50 g/ha (Panels C-D), 100 g/ha (Panels E-F) of Uracilpyridine 20+1% (v/v) MSO. Evaluation and pictures taken 7 Days After Treatment. Application performed on soybean 14 days after sowing. Panels A, C, & E each show plants harboring a PPO trait containing R128A+F420L, R128A+F420I AMTU PPO2 variants as well as untransformed soybean (last column). Panels B, D, & F each show plants harboring a PPO trait containing L397Q+F420V and L397E+F420M AMTU PPO2 variants.
FIG. 11
: Transgenic T4 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed with 25 g/ha (Panels A-B), 50 g/ha (Panels C-D), 100 g/ha (Panels E-F) of Uracilpyridine 21+1% (v/v) MSO. Evaluation and pictures taken 7 Days After Treatment. Application performed on soybean 14 days after sowing. Panels A, C, & E each show plants harboring a PPO trait containing R128A+F420L, R128A+F420I AMTU PPO2 variants as well as untransformed soybean (last column). Panels B, D, & F each show plants harboring a PPO trait containing L397Q+F420V and L397E+F420M AMTU PPO2 variants.
FIG. 12
: Transgenic T4 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed with 25 g/ha (Panels A-B), 50 g/ha (Panels C-D), 100 g/ha (Panels E-F) of Uracilpyridine 22+1% (v/v) MSO. Evaluation and pictures taken 7 Days After Treatment. Application performed on soybean 14 days after sowing. Panels A, C, & E each show plants harboring a PPO trait containing R128A+F420L, R128A+F420I AMTU PPO2 variants as well as untransformed soybean (last column). Panels B, D, & F each show plants harboring a PPO trait containing L397Q+F420V and L397E+F420M AMTU PPO2 variants.
DETAILED DESCRIPTION
The articles “a” and “an” are used herein to refer to one or more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one or more elements.
As used herein, the word “comprising,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.
The inventors of the present invention have found, that the tolerance or resistance of a plant to a uracilpyridine herbicide could be remarkably increased by overexpressing a nucleic acid encoding PPO polypeptides described hereinafter.
a) providing, at said site, a plant that comprises at least one nucleic acid comprising a nucleotide sequence encoding a protoporphyrinogen oxidase (mutated PPO) which is resistant or tolerant to a PPO-inhibiting herbicide as defined hereinafter,
b) applying to said site an effective amount of said herbicide,
The present invention refers to a method for controlling undesired vegetation at a plant cultivation site, the method comprising the steps of:
wherein the PPO inhibiting herbicide is a uracilpyridine of formula (I)
wherein the substituents have the following meanings:
1
2
1
6
3
6
Rhydrogen, NH, C-C-alkyl or C-C-alkynyl;
2
1
6
1
6
Rhydrogen, C-C-alkyl or C-C-haloalkyl;
3
1
6
Rhydrogen or C-C-alkyl;
4
RH or halogen;
5
2
2
3
2
Rhalogen, CN, NO, NH, CFor C(═S)NH;
6
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
RH, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy, C-C-alkylthio, (C-C-alkyl)amino, di(C-C-alkyl)amino, C-C-alkoxy-C-C-alkyl, C-C-alkoxycarbonyl;
7
1
3
1
3
RH, halogen, C-C-alkyl, C-C-alkoxy;
8
9
9
10
11
9
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
—N═CRR, wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
3
6
3
6
1
6
3
6
3
6
1
6
1
4
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, C-C-heterocyclyl-C-C-alkyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
11
9
12
14
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and which carbocyclus is optionally substituted with one to four substituents selected from R;
R, Rindependently of one another are R, or together form a 3- to 7-membered carbocyclus,
ROR, SR, NRR, NROR, NRS(O)Ror NRS(O)NRR, wherein
n 1 to 3;
2
2
1
3
Q CH, O, S, SO, SO, NH or (C-C-alkyl)N;
W O or S;
3
X NH, NCH, O or S;
Y O or S;
1
6
1
6
1
6
1
6
Z phenyl, pyridyl, pyridazinyl, pyrimidinyl or pyrazinyl, each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
including their agriculturally acceptable salts or derivatives, provided the compounds of formula (I) have a carboxyl group.
Sinapis, Lepidium, Galium, Stellaria, Matricaria, Anthemis, Galinsoga, Chenopodium, Urtica, Senecio, Amaranthus, Portulaca, Xanthium, Convolvulus, Ipomoea, Polygonum, Sesbania, Ambrosia, Cirsium, Carduus, Sonchus, Solanum, Rorippa, Rotala, Lindernia, Lamium, Veronica, Abutilon, Emex, Datura, Viola, Galeopsis, Papaver, Centaurea, Trifolium, Ranunculus
Taraxacum
Echinochloa, Setaria, Panicum, Digitaria, Phleum, Poa, Festuca, Eleusine, Brachiaria, Lolium, Bromus, Avena, Cyperus, Sorghum, Agropyron, Cynodon, Monochoria, Fimbristyslis, Sagittaria, Eleocharis, Scirpus, Paspalum, Ischaemum, Sphenoclea, Dactyloctenium, Agrostis, Alopecurus
Apera
The term “control of undesired vegetation” is to be understood as meaning the killing of weeds and/or otherwise retarding or inhibiting the normal growth of the weeds. Weeds, in the broadest sense, are understood as meaning all those plants which grow in locations where they are undesired, e.g. (crop) plant cultivation sites. The weeds of the present invention include, for example, dicotyledonous and monocotyledonous weeds. Dicotyledonous weeds include, but are not limited to, weeds of the genera: , and . Monocotyledonous weeds include, but are not limited to, weeds of the genera: , and . In addition, the weeds of the present invention can include, for example, crop plants that are growing in an undesired location. For example, a volunteer maize plant that is in a field that predominantly comprises soybean plants can be considered a weed, if the maize plant is undesired in the field of soybean plants.
The term “plant” is used in its broadest sense as it pertains to organic material and is intended to encompass eukaryotic organisms that are members of the Kingdom Plantae, examples of which include but are not limited to vascular plants, vegetables, grains, flowers, trees, herbs, bushes, grasses, vines, ferns, mosses, fungi and algae, etc, as well as clones, offsets, and parts of plants used for asexual propagation (e.g. cuttings, pipings, shoots, rhizomes, underground stems, clumps, crowns, bulbs, corms, tubers, rhizomes, plants/tissues produced in tissue culture, etc.). The term “plant” further encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, leaves, roots (including tubers), flowers, florets, fruits, pedicles, peduncles, stamen, anther, stigma, style, ovary, petal, sepal, carpel, root tip, root cap, root hair, leaf hair, seed hair, pollen grain, microspore, cotyledon, hypocotyl, epicotyl, xylem, phloem, parenchyma, endosperm, a companion cell, a guard cell, and any other known organs, tissues, and cells of a plant, and tissues and organs, wherein each of the aforementioned comprise the gene/nucleic acid of interest. The term “plant” also encompasses plant cells, suspension cultures, callus tissue, embryos, meristematic regions, gametophytes, sporophytes, pollen and microspores, again wherein each of the aforementioned comprises the gene/nucleic acid of interest.
Acer
Actinidia
Abelmoschus
Agave sisalana, Agropyron
Agrostis stolonifera, Allium
Amaranthus
Ammophila arenaria, Ananas comosus, Annona
Apium graveolens, Arachis
Artocarpus
Asparagus officinalis, Avena
Avena sativa, Avena fatua, Avena byzantina, Avena fatua
sativa, Avena hybrida
Averrhoa carambola, Bambusa
Benincasa hispida, Bertholletia excelsea, Beta vulgaris, Brassica
Brassica napus, Brassica rapa
Cadaba farinosa, Camellia sinensis, Canna indica, Cannabis sativa, Capsicum
Carex elata, Carica papaya, Carissa macrocarpa, Carya
Carthamus tinctorius, Castanea
Ceiba pentandra, Cichorium endivia, Cinnamomum
Citrullus lanatus, Citrus
Cocos
Coffea
Colocasia esculenta, Cola
Corchorus
Coriandrum sativum, Corylus
Crataegus
Crocus sativus, Cucurbita
Cucumis
Cynara
Daucus carota, Desmodium
Dimocarpus longan, Dioscorea
Diospyros
Echinochloa
Elaeis
Elaeis guineensis, Elaeis oleifera
Eleusine coracana, Eragrostis tef, Erianthus
Eriobotrya japonica, Eucalyptus
Eugenia uniflora, Fagopyrum
Fagus
Festuca arundinacea, Ficus carica, Fortunella
Fragaria
Ginkgo biloba, Glycine
Glycine max, Soja hispida
Soja
Gossypium hirsutum, Helianthus
Helianthus annuus
Hemerocallis fulva, Hibiscus
Hordeum
Hordeum vulgare
Ipomoea batatas, Juglans
Lactuca sativa, Lathyrus
Lens culinaris, Linum usitatissimum, Litchi chinensis, Lotus
Luffa acutangula, Lupinus
Luzula sylvatica, Lycopersicon
Lycopersicon esculentum, Lycopersicon lycopersicum, Lycopersicon pyriforme
Macrotyloma
Malus
Malpighia emarginata, Mammea americana, Mangifera indica, Manihot
Manilkara zapota, Medicago sativa, Melilotus
Mentha
Miscanthus sinensis, Momordica
Morus nigra, Musa
Nicotiana
Olea
Opuntia
Ornithopus
Oryza
Oryza sativa, Oryza latifolia
Panicum miliaceum, Panicum virgatum, Passiflora edulis, Pastinaca sativa, Pennisetum
Persea
Petroselinum crispum, Phalaris arundinacea, Phaseolus
Phleum pratense, Phoenix
Phragmites australis, Physalis
Pinus
Pistacia vera, Pisum
Poa
Populus
Prosopis
Prunus
Psidium
Punica granatum, Pyrus communis, Quercus
Raphanus sativus, Rheum rhabarbarum, Ribes
Ricinus communis, Rubus
Saccharum
Salix
Sambucus
Secale cereale, Sesamum
Sinapis
Solanum
Solanum tuberosum, Solanum integrifolium
Solanum lycopersicum
Sorghum bicolor, Spinacia
Syzygium
Tagetes
Tamarindus
Theobroma cacao, Trifolium
Tripsacum dactyloides, Triticosecale rimpaui, Triticum
Triticum aestivum, Triticum durum, Triticum turgidum, Triticum hybernum, Triticum macha, Triticum sativum, Triticum monococcum
Triticum vulgare
Tropaeolum minus, Tropaeolum majus, Vaccinium
Vicia
Vigna
Viola odorata, Vitis
Zea mays, Zizania palustris
Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including fodder or forage legumes, ornamental plants, food crops, trees or shrubs selected from the list comprising spp., spp., spp., spp., spp., spp., spp., spp, spp., spp. (e.g. var. ), sp., spp. (e.g. ssp. [canola, oilseed rape, turnip rape]), spp., spp., spp., spp., spp., spp., spp., spp., sp., spp., spp., spp., spp., spp., spp., spp., spp., spp., (e.g. ), sp., sp., spp., spp., spp., spp., spp. (e.g. or max), spp. (e.g. ), spp., spp. (e.g. ), spp., spp., spp., spp., spp. (e.g. ), spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp. (e.g. ), sp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., spp., sp., spp., spp., sp., spp. (e.g. or ), spp., spp., spp., indica, spp., spp. (e.g. or ), spp., spp., spp., spp., , Ziziphus spp., amaranth, artichoke, asparagus, broccoli, Brussels sprouts, cabbage, canola, carrot, cauliflower, celery, collard greens, flax, kale, lentil, oilseed rape, okra, onion, potato, rice, soybean, strawberry, sugar beet, sugar cane, sunflower, tomato, squash, tea and algae, amongst others. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include inter ala soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato or tobacco. Further preferably, the plant is a monocotyledonous plant, such as sugarcane. Further preferably, the plant is a cereal, such as rice, maize, wheat, barley, millet, rye, sorghum or oats.
In a preferred embodiment, the plant has been previously produced by a process comprising recombinantly preparing a plant by introducing and over-expressing a wild-type or mutated PPO transgene according to the present invention, as described in greater detail hereinfter.
In another preferred embodiment, the plant has been previously produced by a process comprising in situ mutagenizing plant cells, to obtain plant cells which express a mutated PPO.
As disclosed herein, the nucleic acids of the invention find use in enhancing the herbicide tolerance of plants that comprise in their genomes a gene encoding a herbicide-tolerant wild-type or mutated PPO protein. Such a gene may be an endogenous gene or a transgene, as described hereinafter.
a. Motif 1: SQ[N/K/H]KRYI, wherein the Arg at position 5 within said motif is substituted by any other amino acid;
b. Motif 2: TLGTLFSS, wherein the Leu at position 2 and/or the Gly at position 3, and/or the Leu at position 5 within said motif is substituted by any other amino acid;
c. Motif 3: [F/Y]TTF[V/I]GG, wherein the Phe at position 4 within said motif is substituted by any other amino acid.
Therefore, in another embodiment the present invention refers to a method of increasing or enhancing the uracilpyridine herbicide tolerance or resistance of a plant, the method comprising overexpressing a nucleic acid encoding a PPO polypeptide which comprises one or more of the following motifs 1, 2, and/or 3:
a. the amino acid corresponding to Arg128 of SEQ ID NO: 1 is substituted by any other amino acid.
b. The amino acid corresponding to Gly211 of SEQ ID NO: 1 is substituted by any other amino acid
c. the amino acid corresponding to Leu397 of SEQ ID NO: 1 is substituted by any other amino acid.
d. the amino acid corresponding to Gly398 of SEQ ID NO: 1 is substituted by any other amino acid
e. the amino acid corresponding to Leu400 of SEQ ID NO: 1 is substituted by any other amino acid
f. the amino acid corresponding to Phe420 of SEQ ID NO: 1 is substituted by any other amino acid.
In another embodiment the present invention refers to a method of increasing or enhancing the uracilpyridine herbicide tolerance or resistance of a plant, the method comprising overexpressing a nucleic acid encoding a herbicide resistant or tolerant PPO polypeptide which comprises a variant of the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, which variant comprises one or more of the following substitutions:
In another embodiment the present invention refers to a method of increasing or enhancing the uracilpyridine herbicide tolerance or resistance of a plant, the method comprising overexpressing a nucleic acid encoding a herbicide resistant or tolerant PPO polypeptide which comprises the amino acid sequence of SEQ ID NO: 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, or a variant thereof.
In another embodiment the present invention refers to a method of increasing or enhancing the uracilpyridine herbicide tolerance or resistance of a plant, the method comprising overexpressing a nucleic acid encoding a herbicide resistant or tolerant PPO polypeptide which comprises the amino acid sequence of SEQ ID NO: 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 264.
Bacillus thuringiensis
Additionally, in certain embodiments, the nucleic acids of the present invention can be stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired phenotype. For example, the nucleic acids of the present invention may be stacked with any other polynucleotides encoding polypeptides having pesticidal and/or insecticidal activity, such as, for example, the toxin proteins (described in U.S. Pat. Nos. 5,366,892; 5,747,450; 5,737,514; 5,723,756; 5,593,881; and Geiser et al (1986) Gene 48: 109),
Bacillus thuringiensis
By way of example, polynucleotides that may be stacked with the nucleic acids of the present invention include nucleic acids encoding polypeptides conferring resistance to pests/pathogens such as viruses, nematodes, insects or fungi, and the like. Exemplary polynucleotides that may be stacked with nucleic acids of the invention include polynucleotides encoding: polypeptides having pesticidal and/or insecticidal activity, such as other toxic proteins (described in U.S. Pat. Nos. 5,366,892; 5,747,450; 5,737,514; 5,723,756; 5,593,881; and Geiser et al., (1986) Gene 48:109), lectins (Van Damme et al. (1994) Plant Mol. Biol. 24:825, pentin (described in U.S. Pat. No. 5,981,722), and the like; traits desirable for disease or herbicide resistance (e.g., fumonisin detoxification genes (U.S. Pat. No. 5,792,931); avirulence and disease resistance genes (Jones et al. (1994) Science 266:789; Martin et al., (1993) Science 262:1432; Mindrinos et al. (1994) Cell 78:1089); acetolactate synthase (ALS) mutants that lead to herbicide resistance such as the S4 and/or Hra mutations; glyphosate resistance (e.g., 5-enol-pyrovyl-shikimate-3-phosphate-synthase (EPSPS) gene, described in U.S. Pat. Nos. 4,940,935 and 5,188,642; or the glyphosate N-acetyltransferase (GAT) gene, described in Castle et al. (2004) Science, 304:1151-1154; and in U.S. Patent App. Pub. Nos. 20070004912, 20050246798, and 20050060767)); glufosinate resistance (e.g, phosphinothricin acetyl transferase genes PAT and BAR, described in U.S. Pat. Nos. 5,561,236 and 5,276,268); resistance to herbicides including sulfonyl urea, DHT (2, 4D), and PPO herbicides (e.g., glyphosate acetyl transferase, aryloxy alkanoate dioxygenase, acetolactate synthase, and protoporphyrinogen oxidase); a cytochrome P450 or variant thereof that confers herbicide resistance or tolerance to, inter alia, HPPD herbicides (U.S. patent application Ser. No. 12/156,247; U.S. Pat. Nos. 6,380,465; 6,121,512; 5,349,127; 6,649,814; and 6,300,544; and PCT Patent App. Pub. No. WO2007000077); and traits desirable for processing or process products such as high oil (e.g., U.S. Pat. No. 6,232,529); modified oils (e.g., fatty acid desaturase genes (U.S. Pat. No. 5,952,544; WO 94/11516)); modified starches (e.g., ADPG pyrophosphorylases (AGPase), starch synthases (SS), starch branching enzymes (SBE), and starch debranching enzymes (SDBE)); and polymers or bioplastics (e.g., U.S. Pat. No. 5,602,321; beta-ketothiolase, polyhydroxybutyrate synthase, and acetoacetyl-CoA reductase (Schubert et al. (1988) J. Bacteriol. 170:5837-5847) facilitate expression of polyhydroxyalkanoates (PHAs)); the disclosures of which are herein incorporated by reference.
In a preferred embodiment, the plant comprises at least one additional heterologous nucleic acid comprising a nucleotide sequence encoding a herbicide tolerance enzyme selected, for example, from the group consisting of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), Glyphosate acetyl transferase (GAT), cytochrome P450 monooxygenase, phosphinothricin acetyltransferase (PAT), Acetohydroxyacid synthase (AHAS; EC 4.1.3.18, also known as acetolactate synthase or ALS), hydroxyphenyl pyruvate dioxygenase (HPPD), Phytoene desaturase (PD) and dicamba degrading enzymes as disclosed in WO 02/068607, or phenoxyaceticacid- and phenoxypropionicacid-derivative degrading enzymes as disclosed in WO 2008141154 or WO 2005107437. The combinations generated can also include multiple copies of any one of the polynucleotides of interest.
Generally, the term “herbicide” is used herein to mean an active ingredient that kills, controls or otherwise adversely modifies the growth of plants. The preferred amount or concentration of the herbicide is an “effective amount” or “effective concentration.” By “effective amount” and “effective concentration” is intended an amount and concentration, respectively, that is sufficient to kill or inhibit the growth of a similar, wild-type, plant, plant tissue, plant cell, or host cell, but that said amount does not kill or inhibit as severely the growth of the herbicide-resistant plants, plant tissues, plant cells, and host cells of the present invention. Typically, the effective amount of a herbicide is an amount that is routinely used in agricultural production systems to kill weeds of interest. Such an amount is known to those of ordinary skill in the art. Herbicidal activity is exhibited by herbicides useful for the present invention when they are applied directly to the plant or to the locus of the plant at any stage of growth or before planting or emergence. The effect observed depends upon the plant species to be controlled, the stage of growth of the plant, the application parameters of dilution and spray drop size, the particle size of solid components, the environmental conditions at the time of use, the specific compound employed, the specific adjuvants and carriers employed, the soil type, and the like, as well as the amount of chemical applied. These and other factors can be adjusted as is known in the art to promote non-selective or selective herbicidal action. Generally, it is preferred to apply the herbicide postemergence to relatively immature undesirable vegetation to achieve the maximum control of weeds.
By a “herbicide-tolerant” or “herbicide-resistant” plant, it is intended that a plant that is tolerant or resistant to at least one herbicide at a level that would normally kill, or inhibit the growth of, a normal or wild-type plant. By “herbicide-tolerant wildtype or mutated PPO protein” or “herbicide-resistant wildtype or mutated PPO protein”, it is intended that such a PPO protein displays higher PPO activity, relative to the PPO activity of a wild-type PPO protein, when in the presence of at least one herbicide that is known to interfere with PPO activity and at a concentration or level of the herbicide that is known to inhibit the PPO activity of the wild-type mutated PPO protein. Furthermore, the PPO activity of such a herbicide-tolerant or herbicide-resistant mutated PPO protein may be referred to herein as “herbicide-tolerant” or “herbicide-resistant” PPO activity.
In a particularly preferred embodiment, the herbicides useful for the present invention refer uracilpyridines of formula (I)
wherein the substituents have the following meanings:
1
2
1
6
3
6
Rhydrogen, NH, C-C-alkyl or C-C-alkynyl;
2
1
6
1
6
Rhydrogen, C-C-alkyl or C-C-haloalkyl;
3
1
6
Rhydrogen or C-C-alkyl;
4
RH or halogen;
5
2
2
3
2
Rhalogen, CN, NO, NH, CFor C(═S)NH;
6
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
RH, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy, C-C-alkylthio, (C-C-alkyl)amino, di(C-C-alkyl)amino, C-C-alkoxy-C-C-alkyl, C-C-alkoxycarbonyl;
7
1
3
1
3
RH, halogen, C-C-alkyl, C-C-alkoxy;
8
9
9
10
11
9
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
—N═CRR, wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
3
6
3
6
1
6
3
6
3
6
1
6
1
4
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, C-C-heterocyclyl-C-C-alkyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
11
9
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
R, Rindependently of one another are R, or together form a 3- to 7-membered carbocyclus,
ROR, SR, NRR, NROR, NRS(O)Ror NRS(O)NRR, wherein
n 1 to 3;
2
2
1
3
Q CH, O, S, SO, SO, NH or (C-C-alkyl)N;
W O or S;
3
X NH, NCH, O or S;
Y O or S;
1
6
1
6
1
6
1
6
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
Z phenyl, pyridyl, pyridazinyl, pyrimidinyl or pyrazinyl,
including their agriculturally acceptable salts or derivatives, provided the compounds of formula (I) have a carboxyl group.
If the uracilpyridines of formula (I), the herbicidal compounds B and/or the safeners C as described herein are capable of forming geometrical isomers, for example E/Z isomers, it is possible to use both, the pure isomers and mixtures thereof, in the compositions according to the invention.
If the uracilpyridines of formula (I), the herbicidal compounds B and/or the safeners C as described herein have one or more centres of chirality and, as a consequence, are present as enantiomers or diastereomers, it is possible to use both, the pure enantiomers and diastereomers and their mixtures, in the compositions according to the invention.
Within the substituents of the uracilpyridines of formula (I), instead of hydrogene also the corresponding isotope deuterium can be used.
If the uracilpyridines of formula (I), the herbicidal compounds B and/or the safeners C as described herein have ionizable functional groups, they can also be employed in the form of their agriculturally acceptable salts. Suitable are, in general, the salts of those cations and the acid addition salts of those acids whose cations and anions, respectively, have no adverse effect on the activity of the active compounds.
1
4
1
4
1
4
1
4
1
4
1
4
1
4
1
4
Preferred cations are the ions of the alkali metals, preferably of lithium, sodium and potassium, of the alkaline earth metals, preferably of calcium and magnesium, and of the transition metals, preferably of manganese, copper, zinc and iron, further ammonium and substituted ammonium in which one to four hydrogen atoms are replaced by C-C-alkyl, hydroxy-C-C-alkyl, C-C-alkoxy-C-C-alkyl, hydroxy-C-C-alkoxy-C-C-alkyl, phenyl or benzyl, preferably ammonium, methylammonium, isopropylammonium, dimethylammonium, diethylammonium, diisopropylammonium, trimethylammonium, triethylammonium, tris(isopropyl)ammonium, heptylammonium, dodecylammonium, tetradecylammonium, tetramethylammonium, tetraethylammonium, tetrabutylammonium, 2-hydroxyethylammonium (olamine salt), 2-(2-hydroxyeth-1-oxy)eth-1-ylammonium (diglycolamine salt), di(2-hydroxyeth-1-yl)ammonium (diolamine salt), tris(2-hydroxyethyl)ammonium (trolamine salt), tris(2-hydroxypropyl)ammonium, benzyltrimethylammonium, benzyltriethylammonium, N,N,N-trimethylethanolammonium (choline salt), furthermore phosphonium ions, sulfonium ions, preferably tri(C-C-alkyl)sulfonium, such as trimethylsulfonium, and sulfoxonium ions, preferably tri(C-C-alkyl)sulfoxonium, and finally the salts of polybasic amines such as N,N-bis-(3-aminopropyl)methylamine and diethylenetriamine.
1
4
1
4
1
4
1
4
1
4
1
4
1
4
1
4
Also preferred cations are the ions of the alkali metals, preferably of lithium, sodium and potassium, of the alkaline earth metals, preferably of calcium and magnesium, and of the transition metals, preferably of manganese, copper, zinc and iron, further ammonium and substituted ammonium in which one to four hydrogen atoms are replaced by C-C-alkyl, hydroxy-C-C-alkyl, C-C-alkoxy-C-C-alkyl, hydroxy-C-C-alkoxy-C-C-alkyl, phenyl or benzyl, preferably ammonium, methylammonium, isopropylammonium, dimethylammonium, diisopropylammonium, trimethylammonium, heptylammonium, dodecylammonium, tetradecylammonium, tetramethylammonium, tetraethylammonium, tetrabutylammonium, 2-hydroxyethylammonium (olamine salt), 2-(2-hydroxyeth-1-oxy)eth-1-ylammonium (diglycolamine salt), di(2-hydroxyeth-1-yl)ammonium (diolamine salt), tris(2-hydroxyethyl)ammonium (trolamine salt), tris(2-hydroxypropyl)ammonium, benzyltrimethylammonium, benzyltriethylammonium, N,N,N-trimethylethanolammonium (choline salt), furthermore phosphonium ions, sulfonium ions, preferably tri(C-C-alkyl)sulfonium, such as trimethylsulfonium, and sulfoxonium ions, preferably tri(C-C-alkyl)sulfoxonium, and finally the salts of polybasic amines such as N,N-bis-(3-aminopropyl)methylamine and diethylenetriamine.
1
4
Anions of useful acid addition salts are primarily chloride, bromide, fluoride, iodide, hydrogensulfate, methylsulfate, sulfate, dihydrogenphosphate, hydrogenphosphate, nitrate, bicarbonate, carbonate, hexafluorosilicate, hexafluorophosphate, benzoate and also the anions of C-C-alkanoic acids, preferably formate, acetate, propionate and butyrate.
1
6
1
10
1
10
1
6
1
4
1
4
1
4
1
10
Uracilpyridines of formula (I), herbicidal compounds B and/or safeners C as described herein having a carboxyl group can be employed in the form of the acid, in the form of an agriculturally suitable salt as mentioned above or else in the form of an agriculturally acceptable derivative, for example as amides, such as mono- and di-C-C-alkylamides or arylamides, as esters, for example as allyl esters, propargyl esters, C-C-alkyl esters, alkoxyalkyl esters, tefuryl ((tetrahydrofuran-2-yl)methyl) esters and also as thioesters, for example as C-C-alkylthio esters. Preferred mono- and di-C-C-alkylamides are the methyl and the dimethylamides. Preferred arylamides are, for example, the anilides and the 2-chloroaniides. Preferred alkyl esters are, for example, the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, mexyl (1-methylhexyl), meptyl (1-methylheptyl), heptyl, octyl or isooctyl (2-ethylhexyl) esters. Preferred C-C-alkoxy-C-C-alkyl esters are the straight-chain or branched C-C-alkoxy ethyl esters, for example the 2-methoxyethyl, 2-ethoxyethyl, 2-butoxyethyl (butotyl), 2-butoxypropyl or 3-butoxypropyl ester. An example of a straight-chain or branched C-C-alkylthio ester is the ethylthio ester.
1
14
a
e
n
m
The organic moieties mentioned in the definition of the variables Rto Rand Rto R, are—like the term halogen—collective terms for individual enumerations of the individual group members. The term halogen denotes in each case fluorine, chlorine, bromine or iodine. All hydrocarbon chains, e.g. all alkyl, alkenyl, alkynyl, alkoxy chains, can be straight-chain or branched, the prefix C-Cdenoting in each case the possible number of carbon atoms in the group.
1
3
1
3
1
3
1
3
1
3
3
2
5
3
2
C-C-alkyl and also the C-C-alkyl moieties of di(C-C-alkyl)amino, C-C-alkoxy-C-C-alkyl: for example CH, CH, n-propyl and CH(CH);
1
4
1
4
1
4
3
2
5
3
2
3
2
5
2
3
2
3
3
C-C-alkyl and also the C-C-alkyl moieties of phenyl-C-C-alkyl: for example CH, CH, n-propyl, CH(CH), n-butyl, CH(CH)—CH, CH—CH(CH)and C(CH);
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
4
C-C-alkyl and also the C-C-alkyl moieties of C-C-cyanoalkyl, C-C-alkyoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl-C-C-alkyl: C-C-alkyl as mentioned above, and also, for example, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, 1-ethylpropyl, n-hexyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1-ethyl-1-methylpropyl or 1-ethyl-2-methylpropyl, preferably methyl, ethyl, n-propyl, 1-methylethyl, n-butyl, 1,1-dimethylethyl, n-pentyl or n-hexyl;
1
3
1
3
C-C-haloalkyl: C-C-alkyl as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chlorofluoromethyl, dichlorofluoromethyl, chlorodifluoromethyl, bromomethyl, iodomethyl, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2-iodoethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 2-chloro-2-fluoroethyl, 2-chloro-2,2-difluoroethyl, 2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, pentafluoroethyl, 2-fluoropropyl, 3-fluoropropyl, 2,2-difluoropropyl, 2,3-difluoropropyl, 2-chloropropyl, 3-chloropropyl, 2,3-dichloropropyl, 2-bromopropyl, 3-bromopropyl, 3,3,3-trifluoropropyl, 3,3,3-trichloropropyl, 2,2,3,3,3-pentafluoropropyl, heptafluoropropyl, 1-(fluoromethyl)-2-fluoroethyl, 1-(chloromethyl)-2-chloroethyl, 1-(bromomethyl)-2-bromoethyl;
1
4
1
4
C-C-haloalkyl: C-C-alkyl as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, for example, chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chlorofluoromethyl, dichlorofluoromethyl, chlorodifluoromethyl, bromomethyl, iodomethyl, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2-iodoethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 2-chloro-2-fluoroethyl, 2-chloro-2,2-difluoroethyl, 2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, pentafluoroethyl, 2-fluoropropyl, 3-fluoropropyl, 2,2-difluoropropyl, 2,3-difluoropropyl, 2-chloropropyl, 3-chloropropyl, 2,3-dichloropropyl, 2-bromopropyl, 3-bromopropyl, 3,3,3-trifluoropropyl, 3,3,3-trichloropropyl, 2,2,3,3,3-pentafluoropropyl, heptafluoropropyl, 1-(fluoromethyl)-2-fluoroethyl, 1-(chloromethyl)-2-chloroethyl, 1-(bromomethyl)-2-bromoethyl, 4-fluorobutyl, 4-chlorobutyl, 4-bromobutyl, nonafluorobutyl, 1,1,2,2,-tetrafluoroethyl and 1-trifluoromethyl-1,2,2,2-tetrafluoroethyl;
1
6
1
4
C-C-haloalkyl: C-C-haloalkyl as mentioned above, and also, for example, 5-fluoropentyl, 5-chloropentyl, 5-bromopentyl, 5-iodopentyl, undecafluoropentyl, 6-fluorohexyl, 6-chlorohexyl, 6-bromohexyl, 6-iodohexyl and dodecafluorohexyl;
3
6
3
6
3
6
1
6
3
6
1
6
1
6
3
6
1
6
C-C-alkenyl and also the C-C-alkenyl moieties of C-C-alkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl: for example 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 3-methyl-1-butenyl, 1-methyl-2-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl, 3-methyl-3-butenyl, 1,1-dimethyl-2-propenyl, 1,2-dimethyl-1-propenyl, 1,2-dimethyl-2-propenyl, 1-ethyl-1-propenyl, 1-ethyl-2-propenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 3-methyl-1-pentenyl, 4-methyl-1-pentenyl, 1-methyl-2-pentenyl, 2-methyl-2-pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl, 1-methyl-3-pentenyl, 2-methyl-3-pentenyl, 3-methyl-3-pentenyl, 4-methyl-3-pentenyl, 1-methyl-4-pentenyl, 2-methyl-4-pentenyl, 3-methyl-4-pentenyl, 4-methyl-4-pentenyl, 1,1-dimethyl-2-butenyl, 1,1-dimethyl-3-butenyl, 1,2-dimethyl-1-butenyl, 1,2-dimethyl-2-butenyl, 1,2-dimethyl-3-butenyl, 1,3-dimethyl-1-butenyl, 1,3-dimethyl-2-butenyl, 1,3-dimethyl-3-butenyl, 2,2-dimethyl-3-butenyl, 2,3-dimethyl-1-butenyl, 2,3-dimethyl-2-butenyl, 2,3-dimethyl-3-butenyl, 3,3-dimethyl-1-butenyl, 3,3-dimethyl-2-butenyl, 1-ethyl-1-butenyl, 1-ethyl-2-butenyl, 1-ethyl-3-butenyl, 2-ethyl-1-butenyl, 2-ethyl-2-butenyl, 2-ethyl-3-butenyl, 1,1,2-trimethyl-2-propenyl, 1-ethyl-1-methyl-2-propenyl, 1-ethyl-2-methyl-1-propenyl and 1-ethyl-2-methyl-2-propenyl;
3
6
3
6
3
6
1
6
3
6
C-C-haloalkenyl and also the C-C-haloalkenyl moieties of C-C-haloalkenyloxy-C-C-alkyl: a C-C-alkenyl radical as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, for example 2-chloroprop-2-en-1-yl, 3-chloroprop-2-en-1-yl, 2,3-dichloroprop-2-en-1-yl, 3,3-dichloroprop-2-en-1-yl, 2,3,3-trichloro-2-en-1-yl, 2,3-dichlorobut-2-en-1-yl, 2-bromoprop-2-en-1-yl, 3-bromoprop-2-en-1-yl, 2,3-dibromoprop-2-en-1-yl, 3,3-dibromoprop-2-en-1-yl, 2,3,3-tribromo-2-en-1-yl or 2,3-dibromobut-2-en-1-yl;
3
6
3
6
3
6
1
6
C-C-alkynyl and also the C-C-alkynyl moieties of C-C-alkynyloxycarbonyl-C-C-alkyl: for example 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-methyl-2-butynyl, 1-methyl-3-butynyl, 2-methyl-3-butynyl, 3-methyl-1-butynyl, 1,1-dimethyl-2-propynyl, 1-ethyl-2-propynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 1-methyl-2-pentynyl, 1-methyl-3-pentynyl, 1-methyl-4-pentynyl, 2-methyl-3-pentynyl, 2-methyl-4-pentynyl, 3-methyl-1-pentynyl, 3-methyl-4-pentynyl, 4-methyl-1-pentynyl, 4-methyl-2-pentynyl, 1,1-dimethyl-2-butynyl, 1,1-dimethyl-3-butynyl, 1,2-dimethyl-3-butynyl, 2,2-dimethyl-3-butynyl, 3,3-dimethyl-1-butynyl, 1-ethyl-2-butynyl, 1-ethyl-3-butynyl, 2-ethyl-3-butynyl and 1-ethyl-1-methyl-2-propynyl;
3
6
3
6
C-C-haloalkynyl: a C-C-alkynyl radical as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, for example 1,1-difluoroprop-2-yn-1-yl, 3-chloroprop-2-yn-1-yl, 3-bromoprop-2-yn-1-yl, 3-iodoprop-2-yn-1-yl, 4-fluorobut-2-yn-1-yl, 4-chlorobut-2-yn-1-yl, 1,1-difluorobut-2-yn-1-yl, 4-iodobut-3-yn-1-yl, 5-fluoropent-3-yn-1-yl, 5-iodopent-4-yn-1-yl, 6-fluorohex-4-yn-1-yl or 6-iodohex-5-yn-1-yl;
1
3
1
3
1
3
1
3
1
3
C-C-alkoxy and also the C-C-alkoxy moieties of C-C-alkoxy-C-C-alkyl, C-C-alkoxycarbonyl: for example methoxy, ethoxy, propoxy;
1
4
1
4
1
4
C-C-alkoxy and also the C-C-alkoxy moieties of C-C-alkoxycarbonyl: for example methoxy, ethoxy, propoxy, 1-methylethoxy butoxy, 1-methylpropoxy, 2-methylpropoxy and 1,1-dimethylethoxy;
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
1
6
1
6
1
6
1
4
C-C-alkoxy and also the C-C-alkoxy moieties of C-C-alkyoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl: C-C-alkoxy as mentioned above, and also, for example, pentoxy, 1-methylbutoxy, 2-methylbutoxy, 3-methoxylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, hexoxy, 1-methylpentoxy, 2-methylpentoxy, 3-methylpentoxy, 4-methylpentoxy, 1,1-dimethylbutoxy, 1,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2,2-dimethylbutoxy, 2,3-dimethylbutoxy, 3,3-dimethylbutoxy, 1-ethylbutoxy, 2-ethylbutoxy, 1,1,2-trimethylpropoxy, 1,2,2-trimethylpropoxy, 1-ethyl-1-methylpropoxy and 1-ethyl-2-methylpropoxy.
1
3
1
3
C-C-haloalkoxy: a C-C-alkoxy radical as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, i.e., for example, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chlorodifluoromethoxy, bromodifluoromethoxy, 2-fluoroethoxy, 2-chloroethoxy, 2-bromomethoxy, 2-iodoethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy, 2-chloro-2-fluoroethoxy, 2-chloro-2,2-difluoroethoxy, 2,2-dichloro-2-fluoroethoxy, 2,2,2-trichloroethoxy, pentafluoroethoxy, 2-fluoropropoxy, 3-fluoropropoxy, 2-chloropropoxy, 3-chloropropoxy, 2-bromopropoxy, 3-bromopropoxy, 2,2-difluoropropoxy, 2,3-difluoropropoxy, 2,3-dichloropropoxy, 3,3,3-trifluoropropoxy, 3,3,3-trichloropropoxy, 2,2,3,3,3-pentafluoropropoxy, heptafluoropropoxy, 1-(fluoromethyl)-2-fluoroethoxy, 1-(chloromethyl)-2-chloroethoxy, 1-(bromo-methyl)-2-bromoethoxy;
1
4
1
4
C-C-haloalkoxy: a C-C-alkoxy radical as mentioned above which is partially or fully substituted by fluorine, chlorine, bromine and/or iodine, i.e., for example, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chlorodifluoromethoxy, bromodifluoromethoxy, 2-fluoroethoxy, 2-chloroethoxy, 2-bromomethoxy, 2-iodoethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy, 2-chloro-2-fluoroethoxy, 2-chloro-2,2-difluoroethoxy, 2,2-dichloro-2-fluoroethoxy, 2,2,2-trichloroethoxy, pentafluoroethoxy, 2-fluoropropoxy, 3-fluoropropoxy, 2-chloropropoxy, 3-chloropropoxy, 2-bromopropoxy, 3-bromopropoxy, 2,2-difluoropropoxy, 2,3-difluoropropoxy, 2,3-dichloropropoxy, 3,3,3-trifluoropropoxy, 3,3,3-trichloropropoxy, 2,2,3,3,3-pentafluoropropoxy, heptafluoropropoxy, 1-(fluoromethyl)-2-fluoroethoxy, 1-(chloromethyl)-2-chloroethoxy, 1-(bromo-methyl)-2-bromoethoxy, 4-fluorobutoxy, 4-chlorobutoxy, 4-bromobutoxy and nonafluorobutoxy;
1
6
1
6
1
6
1
6
1
6
1
6
1
4
C-C-haloalkoxy and also the C-C-haloalkoxy moieties of C-C-haloalkoxy-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl: a C-C-haloalkoxy as mentioned above, and also, for example, 5-fluoropentoxy, 5-chloropentoxy, 5-bromopentoxy, 5-iodopentoxy, undecafluoropentoxy, 6-fluorohexoxy, 6-chlorohexoxy, 6-bromohexoxy, 6-iodohexoxy and dodecafluorohexoxy;
1
3
C-C-alkylthio: for example methylthio, ethylthio, propylthio, 1-methylethylthio;
1
4
C-C-alkylthio: for example methylthio, ethylthio, propylthio, 1-methylethylthio, butylthio, 1-methylpropylthio, 2-methylpropylthio and 1,1-dimethylethylthio;
1
6
1
6
1
6
1
6
1
4
C-C-alkylthio and also the C-C-alkylthio moieties of C-C-alkylthio-C-C-alkyl: C-C-alkylthio as mentioned above, and also, for example, pentylthio, 1-methylbutylthio, 2-methylbutylthio, 3-methylbutylthio, 2,2-dimethylpropylthio, 1-ethylpropylthio, hexylthio, 1,1-dimethylpropylthio, 1,2-dimethylpropylthio, 1-methylpentylthio, 2-methylpentylthio, 3-methylpentylthio, 4-methylpentylthio, 1,1-dimethylbutylthio, 1,2-dimethylbutylthio, 1,3-dimethylbutylthio, 2,2-dimethylbutylthio, 2,3-dimethylbutylthio, 3,3-dimethylbutylthio, 1-ethylbutylthio, 2-ethylbutylthio, 1,1,2-trimethylpropylthio, 1,2,2-trimethylpropylthio, 1-ethyl-1-methylpropylthio and 1-ethyl-2-methylpropylthio;
1
6
1
6
1
6
1
6
1
6
C-C-alkylsulfinyl (C-C-alkyl-S(═O)—) and also the C-C-alkylsulfinyl moieties of C-C-alkylsulfinyl-C-C-alkyl: for example methylsulfinyl, ethylsulfinyl, propylsulfinyl, 1-methylethylsulfinyl, butylsulfinyl, 1-methylpropylsulfinyl, 2-methylpropylsulfinyl, 1,1-di-methylethylsulfinyl, pentylsulfinyl, 1-methylbutylsulfinyl, 2-methylbutylsulfinyl, 3-methylbutylsulfinyl, 2,2-dimethylpropylsulfinyl, 1-ethylpropylsulfinyl, 1,1-dimethylpropylsulfinyl, 1,2-dimethylpropylsulfinyl, hexylsulfinyl, 1-methylpentylsulfinyl, 2-methylpentylsulfinyl, 3-methylpentylsulfinyl, 4-methylpentyl-sulfinyl, 1,1-dimethylbutylsulfinyl, 1,2-dimethylbutylsulfinyl, 1,3-dimethylbutyl-sulfinyl, 2,2-dimethylbutylsulfinyl, 2,3-dimethylbutylsulfinyl, 3,3-dimethylbutyl-sulfinyl, 1-ethylbutylsulfinyl, 2-ethylbutylsulfinyl, 1,1,2-trimethylpropylsulfinyl, 1,2,2-trimethylpropylsulfinyl, 1-ethyl-1-methylpropylsulfinyl and 1-ethyl-2-methylpropylsulfinyl;
1
6
1
6
2
1
6
1
6
1
6
C-C-alkylsulfonyl (C-C-alkyl-S(O)—) and also the C-C-alkylsulfonyl moieties of C-C-alkylsulfonyl-C-C-alkyl: for example methylsulfonyl, ethylsulfonyl, propylsulfonyl, 1-methylethylsulfonyl, butylsulfonyl, 1-methylpropylsulfonyl, 2-methyl-propylsulfonyl, 1,1-dimethylethylsulfonyl, pentylsulfonyl, 1-methylbutylsulfonyl, 2-methylbutylsulfonyl, 3-methylbutylsulfonyl, 1,1-dimethylpropylsulfonyl, 1,2-dimethylpropylsulfonyl, 2,2-dimethylpropylsulfonyl, 1-ethylpropylsulfonyl, hexylsulfonyl, 1-methylpentylsulfonyl, 2-methylpentylsulfonyl, 3-methylpentylsulfonyl, 4-methylpentylsulfonyl, 1,1-dimethylbutylsulfonyl, 1,2-dimethylbutylsulfonyl, 1,3-dimethylbutylsulfonyl, 2,2-dimethylbutylsulfonyl, 2,3-dimethylbutylsulfonyl, 3,3-dimethylbutylsulfonyl, 1-ethylbutylsulfonyl, 2-ethylbutylsulfonyl, 1,1,2-trimethylpropylsulfonyl, 1,2,2-trimethylpropylsulfonyl, 1-ethyl-1-methylpropylsulfonyl and 1-ethyl-2-methylpropylsulfonyl;
1
3
(C-C-alkyl)amino: for example methylamino, ethylamino, propylamino, 1-methylethyl-amino;
1
4
(C-C-alkyl)amino: for example methylamino, ethylamino, propylamino, 1-methylethyl-amino, butylamino, 1-methylpropylamino, 2-methylpropylamino or 1,1-dimethylethylamino;
1
6
1
4
(C-C-alkyl)amino: (C-C-alkylamino) as mentioned above, and also, for example, pentylamino, 1-methylbutylamino, 2-methylbutylamino, 3-methylbutylamino, 2,2-dimethylpropylamino, 1-ethylpropylamino, hexylamino, 1,1-dimethylpropylamino, 1,2-dimethylpropylamino, 1-methylpentylamino, 2-methylpentylamino, 3-methylpentylamino, 4-methylpentylamino, 1,1-dimethylbutylamino, 1,2-dimethylbutylamino, 1,3-dimethylbutylamino, 2,2-dimethylbutylamino, 2,3-dimethylbutyl-amino 3,3-dimethylbutylamino, 1-ethylbutylamino, 2-ethylbutylamino, 1,1,2-trimethylpropylamino, 1,2,2-trimethyl-propylamino, 1-ethyl-1-methylpropylamino or 1-ethyl-2-methylpropylamino;
1
6
1
4
di(C-C-alkyl)amino: di(C-C-alkyl)amino as mentioned above, and also, for example, N-methyl-N-pentylamino, N-methyl-N-(1-methylbutyl)amino, N-methyl-N-(2-methylbutyl)amino, N-methyl-N-(3-methylbutyl)amino, N-methyl-N-(2,2-dimethylpropyl)amino, N-methyl-N-(1-ethylpropyl)amino, N-methyl-N-hexylamino, N-methyl-N-(1,1-dimethylpropyl)amino, N-methyl-N-(1,2-dimethylpropyl)amino, N-methyl-N-(1-methylpentyl)amino, N-methyl-N-(2-methylpentyl)amino, N-methyl-N-(3-methylpentyl)amino, N-methyl-N-(4-methylpentyl)amino, N-methyl-N-(1,1-dimethylbutyl)amino, N-methyl-N-(1,2-dimethylbutyl)amino, N-methyl-N-(1,3-dimethylbutyl)amino, N-methyl-N-(2,2-dimethylbutyl)amino, N-methyl-N-(2,3-dimethylbutyl)amino, N-methyl-N-(3,3-dimethylbutyl)amino, N-methyl-N-(1-ethylbutyl)amino, N-methyl-N-(2-ethylbutyl)amino, N-methyl-N-(1,1,2-trimethylpropyl)amino, N-methyl-N-(1,2,2-trimethylpropyl)amino, N-methyl-N-(1-ethyl-1-methylpropyl)amino, N-methyl-N-(1-ethyl-2-methylpropyl)amino, N-ethyl-N-pentylamino, N-ethyl-N-(1-methylbutyl)amino, N-ethyl-N-(2-methylbutyl)amino, N-ethyl-N-(3-methylbutyl)amino, N-ethyl-N-(2,2-dimethylpropyl)amino, N-ethyl-N-(1-ethylpropyl)amino, N-ethyl-N-hexylamino, N-ethyl-N-(1,1-dimethylpropyl)amino, N-ethyl-N-(1,2-dimethylpropyl)amino, N-ethyl-N-(1-methylpentyl)amino, N-ethyl-N-(2-methyl-pentyl)amino, N-ethyl-N-(3-methylpentyl)amino, N-ethyl-N-(4-methylpentyl)amino, N-ethyl-N-(1,1-dimethylbutyl)amino, N-ethyl-N-(1,2-dimethylbutyl)amino, N-ethyl-N-(1,3-dimethylbutyl)amino, N-ethyl-N-(2,2-dimethylbutyl)amino, N-ethyl-N-(2,3-dimethylbutyl)amino, N-ethyl-N-(3,3-dimethylbutyl)amino, N-ethyl-N-(1-ethylbutyl)amino, N-ethyl-N-(2-ethylbutyl)amino, N-ethyl-N-(1,1,2-trimethylpropyl)amino, N-ethyl-N-(1,2,2-trimethylpropyl)amino, N-ethyl-N-(1-ethyl-1-methylpropyl)amino, N-ethyl-N-(1-ethyl-2-methylpropyl)amino, N-propyl-N-pentylamino, N-butyl-N-pentylamino, N,N-dipentylamino, N-propyl-N-hexylamino, N-butyl-N-hexylamino, N-pentyl-N-hexylamino or N,N-dihexylamino;
3
6
3
6
1
6
C-C-cycloalkyl and also the cycloalkyl moieties of C-C-cycloalkyl-C-C-alkyl: monocyclic saturated hydrocarbons having 3 to 6 ring members, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl;
3
6
3
6
1
6
C-C-heterocyclyl and also the heterocyclyl moieties of C-C-heterocyclyl-C-C-alkyl: aliphatic heterocycle having 3 to 6 ring members which, in addition to carbon atoms, contains 1 to 4 nitrogen atoms, or 1 to 3 nitrogen atoms and an oxygen or sulphur atom, or an oxygen or a sulphur atom, for example
Examples of such meanings are:
5- or 6 membered heteroaryl: aromatic heteroaryl having 5 or 6 ring members which, in addition to carbon atoms, contains 1 to 4 nitrogen atoms, or 1 to 3 nitrogen atoms and an oxygen or sulphur atom, or an oxygen or a sulphur atom, for example 5-membered aromatic rings like furyl (for example 2-furyl, 3-furyl), thienyl (for example 2-thienyl, 3-thienyl), pyrrolyl (for example pyrrol-2-yl, pyrrol-3-yl), pyrazolyl (for example pyrazol-3-yl, pyrazol-4-yl), isoxazolyl (for example isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl), isothiazolyl (for example isothiazol-3-yl, isothiazol-4-yl, isothiazol-5-yl), imidazolyl (for example imidazole-2-yl, imidazole-4-yl), oxazolyl (for example oxazol-2-yl, oxazol-4-yl, oxazol-5-yl), thiazolyl (for example thiazol-2-yl, thiazol-4-yl, thiazol-5-yl), oxadiazolyl (for example 1,2,3-oxadiazol-4-yl, 1,2,3-oxadiazol-5-yl, 1,2,4-oxadiazol-3-yl, 1,2,4-oxadiazol-5-yl, 1,3,4-oxadiazol-2-yl), thiadiazolyl (for example 1,2,3-thiadiazol-4-yl, 1,2,3-thiadiazol-5-yl, 1,2,4-thiadiazol-3-yl, 1,2,4-thiadiazol-5-yl, 1,3,4-thiadiazolyl-2-yl), triazolyl (for example 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl); 1-tetrazolyl; 6-membered aromatic rings like pyridyl (for example pyridine-2-yl, pyridine-3-yl, pyridine-4-yl), pyrazinyl (for example pyridazin-3-yl, pyridazin-4-yl), pyrimidinyl (for example pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl), pyrazin-2-yl, triazinyl (for example 1,3,5-triazin-2-yl, 1,2,4-triazin-3-yl, 1,2,4-triazin-5-yl, 1,2,4-triazin-6-yl);
12
3- to 7-membered carbocyclus: a three- to seven-membered monocyclic, saturated, partial unsaturated or aromatic cycle having three to seven ring members which comprises apart from carbon atoms optionally one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—.
three- or four-membered heterocycles like 2-oxetanyl, 3-oxetanyl, 2-thietanyl, 3-thietanyl, 1-azetidinyl, 2-azetidinyl, 1-azetinyl, 2-azetinyl; five-membered saturated heterocycles like 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothienyl, 3-tetrahydrothienyl, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 3-isoxazolidinyl, 4-isoxazolidinyl, 5-isoxazolidinyl, 2-isothiazolidinyl, 3-isothiazolidinyl, 4-isothiazolidinyl, 5-isothiazolidinyl, 1-pyrazolidinyl, 3-pyrazolidinyl, 4-pyrazolidinyl, 5-pyrazolidinyl, 2-oxazolidinyl, 4-oxazolidinyl, 5-oxazolidinyl, 2-thiazolidinyl, 4-thiazolidinyl, 5-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl, 3-oxazolidinyl, 1,2,4-oxadiazolidin-3-yl, 1,2,4-oxadiazolidin-5-yl, 3-thiazolidinyl, 1,2,4-thiadiazolidin-3-yl, 1,2,4-thiadiazolidin-5-yl, 1,2,4-triazolidin-3-yl, 1,2,4-oxadiazolidin-2-yl, 1,2,4-oxadiazolidin-4-yl, 1,3,4-oxadiazolidin-2-yl, 1,2,4-thiadiazolidin-2-yl, 1,2,4-thiadiazolidin-4-yl, 1,3,4-thiadiazolidin-2-yl, 1,2,4-triazolidin-1-yl, 1,3,4-triazolidin-2-yl; six-membered saturated heterocycles like 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 1,3-dioxan-5-yl, 1,4-dioxanyl, 1,3-dithian-5-yl, 1,3-dithianyl, 1,3-oxathian-5-yl, 1,4-oxathianyl, 2-tetrahydropyranyl, 3-tetrahydopyranyl, 4-tetrahydropyranyl, 2-tetrahydrothiopyranyl, 3-tetrahydrothiopyranyl, 4-tetrahydrothiopyranyl, 1-hexahydropyridazinyl, 3-hexahydropyridazinyl, 4-hexahydropyridazinyl, 1-hexahydropyrimidinyl, 2-hexahydropyrimidinyl, 4-hexahydropyrimidinyl, 5-hexahydropyrimidinyl, 1-piperazinyl, 2-piperazinyl, 1,3,5-hexahydrotriazin-1-yl, 1,3,5-hexahydrotriazin-2-yl, 1,2,4-hexahydrotriazin-1-yl, 1,2,4-hexahydrotriazin-3-yl, tetrahydro-1,3-oxazin-1-yl, tetrahydro-1,3-oxazin-2-yl, tetrahydro-1,3-oxazin-6-yl, 1-morpholinyl, 2-morpholinyl, 3-morpholinyl;
The preferred embodiments of the invention mentioned herein below have to be understood as being preferred either independently from each other or in combination with one another.
According to a preferred embodiment of the invention preference is also given to those uracilpyridines of formula (I), wherein the variables, either independently of one another or in combination with one another, have the following meanings:
1
2
1
6
2
1
4
preferably is NHor C-C-alkyl;
2
3
particularly preferred is NHor CH;
1
6
also preferably is C-C-alkyl;
1
4
particularly preferred is C-C-alkyl;
3
especially preferred is CH.
Ris hydrogen, NHor C-C-alkyl;
Preferred are the uracilpyridines of formula (I) wherein
2
1
6
1
6
1
4
1
4
preferably is C-C-alkyl or C-C-haloalkyl;
1
4
more preferred is C-C-haloalkyl;
1
2
particularly preferred is C-C-haloalkyl;
3
especially preferred is CF.
Ris C-C-alkyl or C-C-haloalkyl;
Also preferred are the uracilpyridines of formula (I) wherein
3
1
6
also preferably is C-C-alkyl,
1
4
particularly preferred is C-C-alkyl,
3
especially preferred is CH.
Ris H;
Also preferred are the uracilpyridines of formula (I) wherein
4
particularly preferred is H or F;
especially preferred is H;
also particularly preferred is H or Cl;
especially preferred is Cl;
also particularly preferred is F or Cl;
especially preferred is F.
Ris H, F or Cl;
Also preferred are the uracilpyridines of formula (I) wherein
5
preferably F, Cl, Br or CN;
particularly preferred is F, Cl or CN;
especially preferred is Cl or CN;
more preferred is Cl;
also more preferred is CN;
also especially preferred is F or Cl;
more preferred is F.
Ris halogen or CN;
Also preferred are the uracilpyridines of formula (I) wherein
6
1
3
1
3
1
3
1
3
1
3
1
3
1
3
1
3
particularly preferred is H, C-C-alkyl, C-C-haloalkyl or C-C-alkoxy;
1
3
1
3
especially preferred is H, C-C-alkyl or C-C-alkoxy;
3
3
more preferred is H, CHor OCH.
Ris H, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy or C-C-alkylthio;
Also preferred are the uracilpyridines of formula (I) wherein
7
1
3
3
particularly preferred is H, F or CH;
especially preferred is H.
Ris H, halogen or C-C-alkyl;
Also preferred are the uracilpyridines of formula (I) wherein
8
9
9
10
11
9
10
9
10
11
2
2
9
10
11
9
10
9
10
11
2
2
particularly preferred is OR, NRR, NRS(O)Ror NRS(O)NRR;
9
9
10
9
10
11
2
2
especially preferred OR, NRS(O)Ror NRS(O)NRR;
9
9
10
2
especially preferred is ORor NRS(O)R.
Ris OR, SR, NRR, NRS(O)Ror NRS(O)NRR;
Also preferred are the uracilpyridines of formula (I) wherein
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
3
6
3
6
1
6
3
6
1
4
wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl; C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
14
12
by one to four substituents selected from Ror a 3- to 7-membered carbocyclus, which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted
N═CRR
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
preferably is hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl or C-C-cycloalkyl-C-C-alkyl;
1
6
3
6
3
6
1
6
particularly preferred is hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl or C-C-haloalkyl;
1
6
3
6
3
6
also particularly preferred is hydrogen, C-C-alkyl, C-C-alkenyl or C-C-alkynyl;
1
6
3
6
especially preferred is hydrogen, C-C-alkyl, or C-C-alkynyl;
3
2
5
2
2
2
more preferred is hydrogen, CH, CH, CHCH═CHor CHC═CH;
3
2
5
2
most preferred is hydrogen, CH, CHor CHC═CH.
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxy-carbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
Also preferred are the uracilpyridines of formula (I) wherein
10
1
6
3
6
1
6
particularly preferred is H or C-C-alkyl;
more preferred is H;
1
6
also more preferred is C-C-alkyl.
Ris H, C-C-alkyl or C-C-cycloalkyl;
Also preferred are the uracilpyridines of formula (I) wherein
11
1
6
1
6
1
6
1
6
particularly preferred is H or C-C-alkyl;
more preferred is H;
1
6
also more preferred is C-C-alkyl.
Ris H, C-C-alkyl or C-C-alkoxycarbonyl-C-C-alkyl;
Also preferred are the uracilpyridines of formula (I) wherein
12
1
4
3
particularly preferred is phenyl or CH;
also particularly preferred is phenyl;
1
4
also particularly preferred is C-C-alkyl.
Ris phenyl or C-C-alkyl;
Also preferred are the uracilpyridines of formula (I) wherein
13
1
4
3
particularly preferred is phenyl or CH;
also particularly preferred is phenyl;
1
4
also particularly preferred is C-C-alkyl.
Ris phenyl or C-C-alkyl;
Also preferred are the uracilpyridines of formula (I) wherein
14
1
6
3
particularly preferred is F, Cl or CH;
also particularly preferred is halogen;
1
especially preferred is F or C;
1
6
also particularly preferred is C-C-alkyl;
3
especially preferred is CH.
Ris halogen or C-C-alkyl;
Also preferred are the uracilpyridines of formula (I) wherein
particularly preferred is 2;
also particularly preferred is 1.
n is 1 or 2;
Also preferred are the uracilpyridines of formula (I) wherein
2
1
3
preferably is O or S;
particularly preferred is O.
Q is O, S, SO, SO, NH or (C-C-alkyl)N;
Also preferred are the uracilpyridines of formula (I) wherein
also preferably is S.
W is O,
Also preferred are the uracilpyridines of formula (I) wherein
also preferably is S.
X is O,
Also preferred are the uracilpyridines of formula (I) wherein
also preferably is S.
Y is O,
Also preferred are the uracilpyridines of formula (I) wherein
1
6
1
6
1
6
1
6
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy;
Z is phenyl or pyridyl,
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy;
preferably is phenyl,
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy.
also preferably is pyridyl,
Also preferred are the uracilpyridines of formula (I) wherein
1
6
1
6
1
6
1
6
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy;
Z is phenyl or pyridyl,
1
6
1
6
1
6
1
6
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
preferably is phenyl or pyridyl,
1
6
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen or C-C-alkyl;
particularly preferred is phenyl or pyridyl,
3
each of which is optionally substituted by 1 to 4 substituents selected from the group consisting of F, Cl or CH;
especially preferred is phenyl or pyridyl,
each of which is unsubstituted.
more preferred is phenyl or pyridyl,
Also preferred are the uracilpyridines of formula (I) wherein
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy;
Z is phenyl,
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
preferably is phenyl,
1
6
which is optionally substituted by 1 to 4 substituents selected from the group consisting of halogen or C-C-alkyl;
particularly preferred is phenyl,
1
3
which is optionally substituted by 1 to 4 substituents selected from the group consisting of F, Cor CH;
especially preferred is phenyl
more preferred is unsubstituted phenyl.
Also preferred are the uracilpyridines of formula (I) wherein
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 3 substituents selected from the group consisting of halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy and C-C-haloalkoxy;
Z is pyridyl,
1
6
1
6
1
6
1
6
which is optionally substituted by 1 to 3 substituents selected from the group consisting of halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
preferably is pyridyl,
1
6
which is optionally substituted by 1 to 3 substituents selected from the group consisting of halogen or C-C-alkyl;
particularly preferred is pyridyl,
1
3
which is optionally substituted by 1 to 3 substituents selected from the group consisting of F, Cor CH;
especially preferred is pyridyl,
more preferred is unsubstituted pyridyl.
Also preferred are the uracilpyridines of formula (I) wherein
1
29
Z is selected from the group consisting of Zto Z
Also preferred are the uracilpyridines of formula (I) wherein
wherein
* denotes the point of attachment of Z to X;
** denotes the point of attachment of Z to Q; and
a
b
c
d
e
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
1
6
1
6
1
6
1
6
preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, C, or CH;
more preferred H.
R, R, R, Rand Rindependently of one another are
1
2
3
4
5
6
7
8
9
1
11
12
13
21
1
2
4
5
6
7
8
9
10
11
21
particularly preferred is selected from the group consisting of Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above;
1
4
5
6
7
21
more particularly preferred is selected from the group consisting of Z, Z, Z, Z, Zand Zas defined above;
1
4
5
6
7
especially preferred is selected from the group consisting of Z, Z, Z, Zand Zas defined above;
1
7
more preferred is selected from the group consisting of Zand Zas defined above.
Z is selected from the group consisting of Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above;
Also preferred are the uracilpyridines of formula (I) wherein
1
2
3
4
5
6
7
8
9
10
11
12
13
21
a
b
c
d
e
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy; preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, C, or CH;
more preferred H;
R, R, R, Rand Rindependently of one another are
1
2
4
5
6
7
8
9
10
11
21
a
b
c
d
e
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy; preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, C, or CH;
more preferred H;
particularly preferred is selected from the group consisting of Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are
1
4
5
6
7
21
more particularly preferred is selected from the group consisting of Z, Z, Z, Z, Z, and Zas defined above, wherein
a
b
c
d
e
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy; preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, C, or CH;
more preferred H;
R, R, R, Rand Rindependently of one another are
1
4
5
6
7
especially preferred is selected from the group consisting of Z, Z, Z, Zand Zas defined above, wherein
a
b
c
d
e
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy; preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, Cl, or CH;
more preferred H;
R, R, R, Rand Rindependently of one another are
1
7
more preferred is selected from the group consisting of Zand Zas defined above, wherein
a
b
c
d
e
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy; preferably H, halogen, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-haloalkoxy;
1
6
particularly preferred H, halogen or C-C-alkyl;
3
especially preferred H, F, C, or CH;
more preferred H.
R, R, R, Rand Rindependently of one another are
Z is selected from the group consisting of Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above; wherein
Also preferred are the uracilpyridines of formula (I) wherein
1
1
6
Ris C-C-alkyl,
2
1
4
Ris C-C-haloalkyl,
3
Ris H, and
Y is O.
Also preferred are the uracilpyridines of formula (I) wherein
4
5
Ris F, Cl, Br or CN.
Ris H or F, and
Also preferred are the uracilpyridines of formula (I) wherein
4
Ris H or F, and
5
Ris F, Cl or CN.
Also preferred are the uracilpyridines of formula (I) wherein
6
1
3
1
3
Ris H, C-C-alkyl or C-C-alkoxy, and
7
Ris H.
Also preferred are the uracilpyridines of formula (I) wherein
8
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl or C-C-cycloalkyl-C-C-alkyl; and
10
11
1
6
R, Rare C-C-alkyl.
Ris OR, NRS(O)Ror NRS(O)NRR, wherein
Also preferred are the uracilpyridines of formula (I) wherein
n is 1.
Also preferred are the uracilpyridines of formula (I) wherein
Q, W and X are O.
Also preferred are the uracilpyridines of formula (I) wherein
1
2
1
6
Ris hydrogen, NHor C-C-alkyl;
2
1
6
1
6
Ris C-C-alkyl or C-C-haloalkyl;
3
Ris H;
4
Ris H or halogen;
5
Ris halogen or CN;
6
1
3
1
3
1
3
1
3
1
3
Ris H, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy or C-C-alkylthio;
7
Ris H;
8
9
9
10
11
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
—N═CRR
3
6
3
6
1
6
3
6
1
4
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)amino-carbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
1
6
Ris C-C-alkyl;
11
1
6
Ris H or C-C-alkyl;
12
3
Ris phenyl or CH;
13
3
Ris phenyl or CH;
14
1
6
Ris halogen or C-C-alkyl;
Ris OR, SR, NRR, NRS(O)Ror NRS(O)NRR; wherein
n is 1 or 2;
2
1
3
Q is O, S, SO, SO, NH or (C-C-alkyl)N;
W is O;
X is O;
Y is O;
1
2
3
4
5
6
7
8
9
10
11
12
13
21
a
b
c
d
e
1
6
1
6
1
6
1
6
R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
Z Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above, wherein
Also preferred are the uracilpyridines of formula (I) wherein
1
2
1
4
Ris NHor C-C-alkyl;
2
1
4
1
4
Ris C-C-alkyl or C-C-haloalkyl;
3
Ris H;
4
Ris H or halogen;
5
Ris halogen or CN;
6
1
3
1
3
1
3
Ris H, C-C-alkyl, C-C-haloalkyl or C-C-alkoxy;
7
Ris H;
8
9
10
11
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl or C-C-cycloalkyl-C-C-alkyl;
10
1
6
Ris C-C-alkyl;
11
1
6
Ris H or C-C-alkyl;
ROR, NRR, NRS(O)Ror NRS(O)NRR; wherein
n is 1;
2
1
3
Q is O, S, SO, SO, NH or (C-C-alkyl)N;
W is O;
X is O;
Y is O;
1
2
4
5
6
7
8
9
10
11
21
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Z, Z, Z, Z, Z, Z, Z, Z, Z, Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
particularly preferred are the uracilpyridines of formula (I) wherein
1
2
3
Ris NHor CH;
2
1
4
Ris C-C-haloalkyl;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl, Br or CN;
6
1
3
1
3
Ris H, C-C-alkyl or C-C-alkoxy;
7
Ris H;
8
9
9
10
2
9
1
6
3
6
3
6
1
6
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl or C-C-haloalkyl, and
10
1
6
Ris C-C-alkyl;
Ris ORor NRS(O)R, wherein
n is 1;
Q is O or S;
W is O;
X is O;
Y is O;
1
4
5
6
7
21
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Z, Z, Z, Z, Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
especially preferred are the uracilpyridines of formula (I) wherein
1
2
3
Ris NHor CH;
2
1
4
Ris C-C-haloalkyl;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl or CN;
6
1
3
1
3
Ris H, C-C-alkyl or C-C-alkoxy;
7
Ris H;
8
9
9
10
2
9
1
6
3
6
3
6
1
6
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl or C-C-haloalkyl, and
10
1
6
Ris C-C-alkyl;
Ris ORor NRS(O)R, wherein
n is 1;
Q is O or S;
W is O;
X is O;
Y is O;
1
4
5
6
7
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Z, Z, Z, Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy;
also especially preferred are the uracilpyridines of formula (I) wherein
1
3
Ris CH;
2
3
Ris CF;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl, Br or CN;
6
3
3
Ris H, CHor OCH;
7
Ris H;
8
9
9
10
2
9
1
6
3
6
3
6
Ris hydrogen, C-C-alkyl, C-C-alkenyl, or C-C-alkynyl, and
10
1
6
Ris C-C-alkyl;
Ris ORor NRS(O)R; wherein
n is 1;
Q is O;
W is O;
X is O;
Y is O;
1
7
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy.
more preferred are the uracilpyridines of formula (I) wherein
1
3
Ris CH;
2
3
Ris CF;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl or CN;
6
3
3
Ris H, CHor OCH;
7
Ris H;
8
9
9
10
2
9
1
6
3
6
Ris hydrogen, C-C-alkyl, or C-C-alkynyl, and
10
1
6
Ris C-C-alkyl;
Ris ORor NRS(O)R; wherein
n is 1;
Q is O;
W is O;
X is O;
Y is O;
1
7
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy.
also more preferred are the uracilpyridines of formula (I) wherein
1
3
Ris CH;
2
3
Ris CF;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl, Br or CN;
6
3
3
Ris H, CHor OCH;
7
Ris H;
8
9
9
10
11
9
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
—N═CRR, wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
3
6
3
6
1
6
3
6
3
6
1
6
1
4
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of
12
—N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, C-C-heterocyclyl-C-C-alkyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
11
9
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
R, Rindependently of one another are R, or together form a 3- to 7-membered carbocyclus,
ROR, SR, NRR, NROR, NRS(O)Ror NRS(O)NRR, wherein
n is 1;
Q is O;
W is O;
X is O;
Y is O;
1
7
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy.
Also preferred are the uracilpyridines of formula (I) wherein
1
3
Ris CH;
2
3
Ris CF;
3
Ris H;
4
Ris H, F or Cl;
5
Ris F, Cl or CN;
6
3
3
Ris H, CHor OCH;
7
Ris H;
8
9
9
10
11
9
9
9
10
9
10
11
2
2
9
1
6
3
6
3
6
1
6
3
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
3
6
1
6
3
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
1
6
12
13
12
13
1
4
—N═CRR, wherein Rand Rindependently of one another are H, C-C-alkyl or phenyl;
3
6
3
6
1
6
3
6
3
6
1
6
1
4
14
wherein each cycloalkyl, heterocyclyl, phenyl or heteroaryl ring can be substituted by one to four substituents selected from Ror a 3- to 7-membered carbocyclus,
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of
12
—N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
14
2
1
4
1
4
1
4
1
4
wherein Ris halogen, NO, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy or C-C-alkoxycarbonyl;
C-C-cycloalkyl, C-C-cycloalkyl-C-C-alkyl, C-C-heterocyclyl, C-C-heterocyclyl-C-C-alkyl, phenyl, phenyl-C-C-alkyl or a 5- or 6 membered heteroaryl,
Ris hydrogen, C-C-alkyl, C-C-alkenyl, C-C-alkynyl, C-C-haloalkyl, C-C-haloalkenyl, C-C-haloalkynyl, C-C-cyanoalkyl, C-C-alkoxy-C-C-alkyl, C-C-alkoxy-C-C-alkoxy-C-C-alkyl, di(C-C-alkoxy)C-C-alkyl, C-C-haloalkoxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkyl, C-C-haloalkenyloxy-C-C-alkyl, C-C-alkenyloxy-C-C-alkoxy-C-C-alkyl, C-C-alkylthio-C-C-alkyl, C-C-alkylsulfinyl-C-C-alkyl, C-C-alkylsulfonyl-C-C-alkyl, C-C-alkylcarbonyl-C-C-alkyl, C-C-alkoxycarbonyl-C-C-alkyl, C-C-haloalkoxycarbonyl-C-C-alkyl, C-C-alkenyloxycarbonyl-C-C-alkyl, C-C-alkynyloxycarbonyl-C-C-alkyl, amino, (C-C-alkyl)amino, di(C-C-alkyl)amino, (C-C-alkylcarbonyl)amino, amino-C-C-alkyl, (C-C-alkyl)amino-C-C-alkyl, di(C-C-alkyl)amino-C-C-alkyl, aminocarbonyl-C-C-alkyl, (C-C-alkyl)aminocarbonyl-C-C-alkyl, di(C-C-alkyl)aminocarbonyl-C-C-alkyl,
10
11
9
12
which carbocyclus optionally has in addition to carbon atoms one or two ring members selected from the group consisting of —N(R)—, —N═N—, —C(═O)—, —O— and —S—, and
14
which carbocyclus is optionally substituted with one to four substituents selected from R;
R, Rindependently of one another are R, or together form a 3- to 7-membered carbocyclus,
ROR, SR, NRR, NROR, NRS(O)Ror NRS(O)NRR, wherein
n is 1;
Q is O;
W is O;
X is O;
Y is O;
1
7
a
b
c
d
e
1
6
1
6
1
6
1
6
Z is selected from the group consisting of Zand Zas defined above, wherein R, R, R, Rand Rindependently of one another are H, halogen, CN, C-C-alkyl, C-C-haloalkyl, C-C-alkoxy, C-C-haloalkoxy.
Also preferred are the uracilpyridines of formula (I) wherein
1
2
3
7
a
b
c
d
3
3
Particular preference is given to uracilpyrimidines of formula (I.a) (corresponds to formula (I) wherein Ris CH, Ris CF, Ris H, Ris H, n is 1, Q, W, X and Y are O, and Z is Z-1 as defined, wherein R, R, Rand Rare H:
4
5
6
8
wherein the variables R, R, Rand Rhave the meanings, in particular the preferred meanings, as defined above.
4
5
6
8
Special preference is given to the compounds of the formulae (I.a.1) to (I.a.672), preferably (I.a.1) to (I.a.504), of Table A, where the definitions of the variables R, R, Rand Rare of particular importance for the compounds according to the invention not only in combination with one another but in each case also on their own:
TABLE A
No.
R<sup>4</sup>
R<sup>5</sup>
R<sup>6</sup>
R<sup>8</sup>
I.a.1
H
F
H
OH
I.a.2
H
F
H
OCH<sub>3</sub>
I.a.3
H
F
H
OC<sub>2</sub>H<sub>5</sub>
I.a.4
H
F
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.5
H
F
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.6
H
F
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.7
H
F
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.8
H
F
H
OCH<sub>2</sub>C≡CH
I.a.9
H
F
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.10
H
F
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.11
H
F
H
OC<sub>6</sub>H<sub>5</sub>
I.a.12
H
F
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.13
H
F
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.14
H
F
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.15
H
F
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.16
H
F
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.17
H
F
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.18
H
F
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.19
H
F
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.20
H
F
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.21
H
F
H
OCH<sub>2</sub>-cyclopropyl
I.a.22
H
F
H
OCH<sub>2</sub>-cyclobutyl
I.a.23
H
F
H
SCH<sub>3</sub>
I.a.24
H
F
H
SC<sub>2</sub>H<sub>5</sub>
I.a.25
H
F
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.26
H
F
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.27
H
F
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.28
H
F
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.29
H
F
CH<sub>3</sub>
OH
I.a.30
H
F
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.31
H
F
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.32
H
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.33
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.34
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.35
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.36
H
F
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.37
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.38
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.39
H
F
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.40
H
F
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.41
H
F
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.42
H
F
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.43
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.44
H
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.45
H
F
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.46
H
F
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.47
H
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.48
H
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.49
H
F
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.50
H
F
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.51
H
F
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.52
H
F
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.53
H
F
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.54
H
F
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.55
H
F
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.56
H
F
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.57
H
F
OCH<sub>3</sub>
OH
I.a.58
H
F
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.59
H
F
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.60
H
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.61
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.62
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.63
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.64
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.65
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.66
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.67
H
F
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.68
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.69
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.70
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.71
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.72
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.73
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.74
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.75
H
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.76
H
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.77
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.78
H
F
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.79
H
F
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.80
H
F
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.81
H
F
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.82
H
F
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.83
H
F
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.84
H
F
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.85
H
Cl
H
OH
I.a.86
H
Cl
H
OCH<sub>3</sub>
I.a.87
H
Cl
H
OC<sub>2</sub>H<sub>5</sub>
I.a.88
H
Cl
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.89
H
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.90
H
Cl
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.91
H
Cl
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.92
H
Cl
H
OCH<sub>2</sub>C≡CH
I.a.93
H
Cl
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.94
H
Cl
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.95
H
Cl
H
OC<sub>6</sub>H<sub>5</sub>
I.a.96
H
Cl
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.97
H
Cl
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.98
H
Cl
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.99
H
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.100
H
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.101
H
Cl
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.102
H
Cl
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.103
H
Cl
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.104
H
Cl
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.105
H
Cl
H
OCH<sub>2</sub>-cyclopropyl
I.a.106
H
Cl
H
OCH<sub>2</sub>-cyclobutyl
I.a.107
H
Cl
H
SCH<sub>3</sub>
I.a.108
H
Cl
H
SC<sub>2</sub>H<sub>5</sub>
I.a.109
H
Cl
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.110
H
Cl
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.111
H
Cl
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.112
H
Cl
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.113
H
Cl
CH<sub>3</sub>
OH
I.a.114
H
Cl
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.115
H
Cl
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.116
H
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.117
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.118
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.119
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.120
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.121
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.122
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.123
H
Cl
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.124
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.125
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.126
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.127
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.128
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.129
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.130
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.131
H
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.132
H
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.133
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.134
H
Cl
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.135
H
Cl
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.136
H
Cl
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.137
H
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.138
H
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.139
H
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.140
H
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.141
H
Cl
OCH<sub>3</sub>
OH
I.a.142
H
Cl
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.143
H
Cl
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.144
H
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.145
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.146
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.147
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.148
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.149
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.150
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.151
H
Cl
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.152
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.153
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.154
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.155
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.156
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.157
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.158
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.159
H
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.160
H
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.161
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.162
H
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.163
H
Cl
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.164
H
Cl
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.165
H
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.166
H
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.167
H
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.168
H
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.169
H
CN
H
OH
I.a.170
H
CN
H
OCH<sub>3</sub>
I.a.171
H
CN
H
OC<sub>2</sub>H<sub>5</sub>
I.a.172
H
CN
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.173
H
CN
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.174
H
CN
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.175
H
CN
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.176
H
CN
H
OCH<sub>2</sub>C≡CH
I.a.177
H
CN
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.178
H
CN
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.179
H
CN
H
OC<sub>6</sub>H<sub>5</sub>
I.a.180
H
CN
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.181
H
CN
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.182
H
CN
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.183
H
CN
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.184
H
CN
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.185
H
CN
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.186
H
CN
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.187
H
CN
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.188
H
CN
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.189
H
CN
H
OCH<sub>2</sub>-cyclopropyl
I.a.190
H
CN
H
OCH<sub>2</sub>-cyclobutyl
I.a.191
H
CN
H
SCH<sub>3</sub>
I.a.192
H
CN
H
SC<sub>2</sub>H<sub>5</sub>
I.a.193
H
CN
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.194
H
CN
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.195
H
CN
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.196
H
CN
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.197
H
CN
CH<sub>3</sub>
OH
I.a.198
H
CN
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.199
H
CN
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.200
H
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.201
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.202
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.203
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.204
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.205
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.206
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.207
H
CN
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.208
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.209
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.210
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.211
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.212
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.213
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.214
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.215
H
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.216
H
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.217
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.218
H
CN
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.219
H
CN
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.220
H
CN
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.221
H
CN
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.222
H
CN
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.223
H
CN
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.224
H
CN
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.225
H
CN
OCH<sub>3</sub>
OH
I.a.226
H
CN
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.227
H
CN
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.228
H
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.229
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.230
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.231
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.232
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.233
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.234
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.235
H
CN
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.236
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.237
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.238
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.239
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.240
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.241
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.242
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.243
H
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.244
H
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.245
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.246
H
CN
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.247
H
CN
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.248
H
CN
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.249
H
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.250
H
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.251
H
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.252
H
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.253
F
F
H
OH
I.a.254
F
F
H
OCH<sub>3</sub>
I.a.255
F
F
H
OC<sub>2</sub>H<sub>5</sub>
I.a.256
F
F
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.257
F
F
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.258
F
F
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.259
F
F
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.260
F
F
H
OCH<sub>2</sub>C≡CH
I.a.261
F
F
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.262
F
F
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.263
F
F
H
OC<sub>6</sub>H<sub>5</sub>
I.a.264
F
F
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.265
F
F
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.266
F
F
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.267
F
F
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.268
F
F
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.269
F
F
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.270
F
F
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.271
F
F
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.272
F
F
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.273
F
F
H
OCH<sub>2</sub>-cyclopropyl
I.a.274
F
F
H
OCH<sub>2</sub>-cyclobutyl
I.a.275
F
F
H
SCH<sub>3</sub>
I.a.276
F
F
H
SC<sub>2</sub>H<sub>5</sub>
I.a.277
F
F
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.278
F
F
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.279
F
F
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.280
F
F
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.281
F
F
CH<sub>3</sub>
OH
I.a.282
F
F
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.283
F
F
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.284
F
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.285
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.286
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.287
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.288
F
F
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.289
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.290
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.291
F
F
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.292
F
F
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.293
F
F
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.294
F
F
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.295
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.296
F
F
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.297
F
F
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.298
F
F
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.299
F
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.300
F
F
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.301
F
F
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.302
F
F
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.303
F
F
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.304
F
F
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.305
F
F
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.306
F
F
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.307
F
F
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.308
F
F
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.309
F
F
OCH<sub>3</sub>
OH
I.a.310
F
F
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.311
F
F
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.312
F
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.313
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.314
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.315
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.316
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.317
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.318
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.319
F
F
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.320
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.321
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.322
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.323
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.324
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.325
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.326
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.327
F
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.328
F
F
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.329
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.330
F
F
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.331
F
F
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.332
F
F
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.333
F
F
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.334
F
F
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.335
F
F
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.336
F
F
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.337
F
Cl
H
OH
I.a.338
F
Cl
H
OCH<sub>3</sub>
I.a.339
F
Cl
H
OC<sub>2</sub>H<sub>5</sub>
I.a.340
F
Cl
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.341
F
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.342
F
Cl
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.343
F
Cl
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.344
F
Cl
H
OCH<sub>2</sub>C≡CH
I.a.345
F
Cl
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.346
F
Cl
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.347
F
Cl
H
OC<sub>6</sub>H<sub>5</sub>
I.a.348
F
Cl
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.349
F
Cl
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.350
F
Cl
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.351
F
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.352
F
Cl
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.353
F
Cl
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.354
F
Cl
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.355
F
Cl
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.356
F
Cl
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.357
F
Cl
H
OCH<sub>2</sub>-cyclopropyl
I.a.358
F
Cl
H
OCH<sub>2</sub>-cyclobutyl
I.a.359
F
Cl
H
SCH<sub>3</sub>
I.a.360
F
Cl
H
SC<sub>2</sub>H<sub>5</sub>
I.a.361
F
Cl
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.362
F
Cl
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.363
F
Cl
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.364
F
Cl
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.365
F
Cl
CH<sub>3</sub>
OH
I.a.366
F
Cl
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.367
F
Cl
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.368
F
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.369
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.370
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.371
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.372
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.373
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.374
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.375
F
Cl
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.376
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.377
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.378
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.379
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.380
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.381
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.382
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.383
F
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.384
F
Cl
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.385
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.386
F
Cl
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.387
F
Cl
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.388
F
Cl
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.389
F
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.390
F
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.391
F
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.392
F
Cl
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.393
F
Cl
OCH<sub>3</sub>
OH
I.a.394
F
Cl
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.395
F
Cl
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.396
F
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.397
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.398
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.399
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.400
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.401
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.402
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.403
F
Cl
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.404
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.405
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.406
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.407
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.408
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.409
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.410
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.411
F
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.412
F
Cl
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.413
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.414
F
Cl
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.415
F
Cl
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.416
F
Cl
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.417
F
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.418
F
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.419
F
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.420
F
Cl
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.421
F
CN
H
OH
I.a.422
F
CN
H
OCH<sub>3</sub>
I.a.423
F
CN
H
OC<sub>2</sub>H<sub>5</sub>
I.a.424
F
CN
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.425
F
CN
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.426
F
CN
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.427
F
CN
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.428
F
CN
H
OCH<sub>2</sub>C≡CH
I.a.429
F
CN
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.430
F
CN
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.431
F
CN
H
OC<sub>6</sub>H<sub>5</sub>
I.a.432
F
CN
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.433
F
CN
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.434
F
CN
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.435
F
CN
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.436
F
CN
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.437
F
CN
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.438
F
CN
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.439
F
CN
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.440
F
CN
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.441
F
CN
H
OCH<sub>2</sub>-cyclopropyl
I.a.442
F
CN
H
OCH<sub>2</sub>-cyclobutyl
I.a.443
F
CN
H
SCH<sub>3</sub>
I.a.444
F
CN
H
SC<sub>2</sub>H<sub>5</sub>
I.a.445
F
CN
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.446
F
CN
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.447
F
CN
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.448
F
CN
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.449
F
CN
CH<sub>3</sub>
OH
I.a.450
F
CN
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.451
F
CN
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.452
F
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.453
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.454
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.455
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.456
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.457
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.458
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.459
F
CN
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.460
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.461
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.462
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.463
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.464
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.465
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.466
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.467
F
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.468
F
CN
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.469
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.470
F
CN
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.471
F
CN
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.472
F
CN
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.473
F
CN
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.474
F
CN
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.475
F
CN
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.476
F
CN
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.477
F
CN
OCH<sub>3</sub>
OH
I.a.478
F
CN
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.479
F
CN
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.480
F
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.481
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.482
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.483
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.484
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.485
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.486
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.487
F
CN
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.488
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.489
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.490
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.491
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.492
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.493
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.494
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.495
F
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.496
F
CN
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.497
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.498
F
CN
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.499
F
CN
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.500
F
CN
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.501
F
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.502
F
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.503
F
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.504
F
CN
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.505
H
Br
H
OH
I.a.506
H
Br
H
OCH<sub>3</sub>
I.a.507
H
Br
H
OC<sub>2</sub>H<sub>5</sub>
I.a.508
H
Br
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.509
H
Br
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.510
H
Br
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.511
H
Br
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.512
H
Br
H
OCH<sub>2</sub>C≡CH
I.a.513
H
Br
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.514
H
Br
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.515
H
Br
H
OC<sub>6</sub>H<sub>5</sub>
I.a.516
H
Br
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.517
H
Br
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.518
H
Br
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.519
H
Br
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.520
H
Br
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.521
H
Br
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.522
H
Br
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.523
H
Br
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.524
H
Br
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.525
H
Br
H
OCH<sub>2</sub>-cyclopropyl
I.a.526
H
Br
H
OCH<sub>2</sub>-cyclobutyl
I.a.527
H
Br
H
SCH<sub>3</sub>
I.a.528
H
Br
H
SC<sub>2</sub>H<sub>5</sub>
I.a.529
H
Br
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.530
H
Br
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.531
H
Br
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.532
H
Br
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.533
H
Br
CH<sub>3</sub>
OH
I.a.534
H
Br
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.535
H
Br
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.536
H
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.537
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.538
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.539
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.540
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.541
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.542
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.543
H
Br
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.544
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.545
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.546
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.547
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.548
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.549
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.550
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.551
H
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.552
H
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.553
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.554
H
Br
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.555
H
Br
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.556
H
Br
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.557
H
Br
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.558
H
Br
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.559
H
Br
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.560
H
Br
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.561
H
Br
OCH<sub>3</sub>
OH
I.a.562
H
Br
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.563
H
Br
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.564
H
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.565
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.566
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.567
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.568
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.569
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.570
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.571
H
Br
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.572
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.573
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.574
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.575
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.576
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.577
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.578
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.579
H
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.580
H
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.581
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.582
H
Br
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.583
H
Br
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.584
H
Br
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.585
H
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.586
H
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.587
H
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.588
H
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.589
F
Br
H
OH
I.a.590
F
Br
H
OCH<sub>3</sub>
I.a.591
F
Br
H
OC<sub>2</sub>H<sub>5</sub>
I.a.592
F
Br
H
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.593
F
Br
H
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.594
F
Br
H
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.595
F
Br
H
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.596
F
Br
H
OCH<sub>2</sub>C≡CH
I.a.597
F
Br
H
OCH<sub>2</sub>CF<sub>3</sub>
I.a.598
F
Br
H
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.599
F
Br
H
OC<sub>6</sub>H<sub>5</sub>
I.a.600
F
Br
H
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.601
F
Br
H
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.602
F
Br
H
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.603
F
Br
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.604
F
Br
H
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.605
F
Br
H
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.606
F
Br
H
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.607
F
Br
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.608
F
Br
H
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.609
F
Br
H
OCH<sub>2</sub>-cyclopropyl
I.a.610
F
Br
H
OCH<sub>2</sub>-cyclobutyl
I.a.611
F
Br
H
SCH<sub>3</sub>
I.a.612
F
Br
H
SC<sub>2</sub>H<sub>5</sub>
I.a.613
F
Br
H
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.614
F
Br
H
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.615
F
Br
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.616
F
Br
H
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.617
F
Br
CH<sub>3</sub>
OH
I.a.618
F
Br
CH<sub>3</sub>
OCH<sub>3</sub>
I.a.619
F
Br
CH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.620
F
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.621
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.622
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.623
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.624
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.625
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.626
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.627
F
Br
CH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.628
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.629
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.630
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.631
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.632
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.633
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.634
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.635
F
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.636
F
Br
CH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.637
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.638
F
Br
CH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.639
F
Br
CH<sub>3</sub>
SCH<sub>3</sub>
I.a.640
F
Br
CH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.641
F
Br
CH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.642
F
Br
CH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.643
F
Br
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.644
F
Br
CH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
I.a.645
F
Br
OCH<sub>3</sub>
OH
I.a.646
F
Br
OCH<sub>3</sub>
OCH<sub>3</sub>
I.a.647
F
Br
OCH<sub>3</sub>
OC<sub>2</sub>H<sub>5</sub>
I.a.648
F
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)<sub>2</sub>
I.a.649
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>CH<sub>3</sub>
I.a.650
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.651
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH═CH<sub>2</sub>
I.a.652
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>C≡CH
I.a.653
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CF<sub>3</sub>
I.a.654
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CHF<sub>2</sub>
I.a.655
F
Br
OCH<sub>3</sub>
OC<sub>6</sub>H<sub>5</sub>
I.a.656
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(C<sub>6</sub>H<sub>5</sub>)
I.a.657
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>3</sub>
I.a.658
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.659
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>3</sub>
I.a.660
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>CH<sub>2</sub>OCH<sub>2</sub>CH<sub>3</sub>
I.a.661
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>3</sub>
I.a.662
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.663
F
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>3</sub>
I.a.664
F
Br
OCH<sub>3</sub>
OCH(CH<sub>3</sub>)(CO)OCH<sub>2</sub>CH<sub>3</sub>
I.a.665
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclopropyl
I.a.666
F
Br
OCH<sub>3</sub>
OCH<sub>2</sub>-cyclobutyl
I.a.667
F
Br
OCH<sub>3</sub>
SCH<sub>3</sub>
I.a.668
F
Br
OCH<sub>3</sub>
SC<sub>2</sub>H<sub>5</sub>
I.a.669
F
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>CH<sub>3</sub>
I.a.670
F
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>CH(CH<sub>3</sub>)<sub>2</sub>
I.a.671
F
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)<sub>2</sub>
I.a.672
F
Br
OCH<sub>3</sub>
NHSO<sub>2</sub>N(CH<sub>3</sub>)[CH(CH<sub>3</sub>)<sub>2</sub>]
Also preferred are the uracilpyridines of formula (I.b), preferably the uracilpyridines of formulae (I.b.1) to (I.b.672), particularly preferred the uracilpyridines of formulae (I.b.1) to (I.b.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Q is S:
a
b
d
e
Also preferred are the uracilpyridines of formula (I.c), preferably the uracilpyridines of formulae (I.c.1) to (I.c.672), particularly preferred the uracilpyridines of formulae (I.c.1) to (I.c.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-2, wherein R, R, Rand Rare H:
a
b
d
e
Also preferred are the uracilpyridines of formula (I.d), preferably the uracilpyridines of formulae (I.d.1) to (I.d.672), particularly preferred the uracilpyridines of formulae (I.d.1) to (I.d.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-3, wherein R, R, Rand Rare H:
b
c
d
Also preferred are the uracilpyridines of formula (I.e), preferably the uracilpyridines of formulae (I.e.1) to (I.e.672), particularly preferred the uracilpyridines of formulae (I.e.1) to (I.e.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-4, wherein R, Rand Rare H:
a
c
d
Also preferred are the uracilpyridines of formula (I.f), preferably the uracilpyridines of formulae (I.f.1) to (I.f.672), particularly preferred the uracilpyridines of formulae (I.f.1) to (I.f.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-5, wherein R. Rand Rare H:
a
b
d
Also preferred are the uracilpyridines of formula (I.g), preferably the uracilpyridines of formulae (I.g.1) to (I.g.672),particularly preferred the uracilpyridines of formulae (I.g.1) to (I.g.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-6, wherein R, Rand Rare H:
a
b
c
Also preferred are the uracilpyridines of formula (I.h), preferably the uracilpyridines of formulae (I.h.1) to (I.h.672), particularly preferred the uracilpyridines of formulae (I.h.1) to (I.h.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-7, wherein R, Rand Rare H:
a
b
c
Also preferred are the uracilpyridines of formula (I.i), preferably the uracilpyridines of formulae (I.i.1) to (I.i.672), particularly preferred the uracilpyridines of formulae (I.i.1) to (I.i.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-7, wherein R, Rand Rare H, and Q is S:
b
c
e
Also preferred are the uracilpyridines of formula (I.k), preferably the uracilpyridines of formulae (I.k.1) to (I.k.672), particularly preferred the uracilpyridines of formulae (I.k.1) to (I.k.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-8, wherein R, Rand Rare H:
a
c
e
Also preferred are the uracilpyridines of formula (I.l), preferably the uracilpyridines of formulae (I.l.1) to (I.l.672), particularly preferred the uracilpyridines of formulae (I.l.1) to (I.l.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-9, wherein R, Rand Rare H:
a
b
e
Also preferred are the uracilpyridines of formula (I.m), preferably the uracilpyridines of formulae (I.m.1) to (I.m.672), particularly preferred the uracilpyridines of formulae (I.m.1) to (I.m.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-10, wherein R, Rand Rare H:
a
b
c
Also preferred are the uracilpyridines of formula (I.n), preferably the uracilpyridines of formulae (I.n.1) to (I.n.672), particularly preferred the uracilpyridines of formulae (I.n.1) to (I.n.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-11, wherein R, Rand Rare H:
b
d
e
Also preferred are the uracilpyridines of formula (I.o), preferably the uracilpyridines of formulae (I.o.1) to (I.o.672), particularly preferred the uracilpyridines of formulae (I.o.1) to (I.o.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-12, wherein R, Rand Rare H:
a
d
e
Also preferred are the uracilpyridines of formula (I.p), preferably the uracilpyridines of formulae (I.p.1) to (I.p.672), particularly preferred the uracilpyridines of formulae (I.p.1) to (I.p.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-13, wherein R, Rand Rare H:
a
c
Also preferred are the uracilpyridines of formula (I.q), preferably the uracilpyridines of formulae (I.q.1) to (I.q.672), particularly preferred the uracilpyridines of formulae (I.q.1) to (I.q.504), which differ from the corresponding uracilpyridines of formulae (I.a.1) to (I.a.672) only in that Z is Z-21, wherein Rand Rare H:
To widen the spectrum of action and to achieve synergistic effects, the uracilpyridines of formula (I) may be mixed with a large number of representatives of other herbicidal or growth-regulating active ingredient groups and then applied concomitantly. Suitable components for mixtures are, for example,
herbicides from the classes of the acetamides, amides, aryloxyphenoxypropionates, benzamides, benzofuran, benzoic acids, benzothiadiazinones, bipyridylium, carbamates, chloroacetamides, chlorocarboxylic acids, cyclohexanediones, dinitroanilines, dinitrophenol, diphenyl ether, glycines, imidazolinones, isoxazoles, isoxazolidinones, nitriles, N-phenylphthalimides, oxadiazoles, oxazolidinediones, oxyacetamides, phenoxycarboxylic acids, phenylcarbamates, phenylpyrazoles, phenylpyrazolines, phenylpyridazines, phosphinic acids, phosphoroamidates, phosphorodithioates, phthalamates, pyrazoles, pyridazinones, pyridines, pyridinecarboxylic acids, pyridinecarboxamides, pyrimidinediones, pyrimidinyl(thio)benzoates, quinolinecarboxylic acids, semicarbazones, sulfonylaminocarbonyltriazolinones, sulfonylureas, tetrazolinones, thiadiazoles, thiocarbamates, triazines, triazinones, triazoles, triazolinones, triazolocarboxamides, triazolopyrimidines, triketones, uracils, ureas.
It may furthermore be beneficial to apply the uracilpyridines of formula (I) alone or in combination with other herbicides, or else in the form of a mixture with other crop protection agents, for example together with agents for controlling pests or phytopathogenic fungi or bacteria. Also of interest is the miscibility with mineral salt solutions, which are employed for treating nutritional and trace element deficiencies. Other additives such as non-phytotoxic oils and oil concentrates may also be added.
In one embodiment of the present invention the compositions according to the present invention comprise at least one uracilpyridine of formula (I) (compound A) and at least one further active compound selected from herbicides B, preferably herbicides B of class b1) to b15), and safeners C (compound C).
In another embodiment of the present invention the compositions according to the present invention comprise at least one uracilpyridine of formula (I) and at least one further active compound B (herbicide B).
b1) lipid biosynthesis inhibitors;
b2) acetolactate synthase inhibitors (ALS inhibitors);
b3) photosynthesis inhibitors;
b4) protoporphyrinogen-IX oxidase inhibitors,
b5) bleacher herbicides;
b6) enolpyruvyl shikimate 3-phosphate synthase inhibitors (EPSP inhibitors);
b7) glutamine synthetase inhibitors;
b8) 7,8-dihydropteroate synthase inhibitors (DHP inhibitors);
b9) mitosis inhibitors;
b10) inhibitors of the synthesis of very long chain fatty acids (VLCFA inhibitors);
b11) cellulose biosynthesis inhibitors;
b12) decoupler herbicides;
b13) auxinic herbicides;
b14) auxin transport inhibitors; and
b15) other herbicides selected from the group consisting of bromobutide, chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, dalapon, dazomet, difenzoquat, difenzoquat-metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, indaziflam, maleic hydrazide, mefluidide, metam, methiozolin (CAS 403640-27-7), methyl azide, methyl bromide, methyl-dymron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine, triaziflam, tridiphane and 6-chloro-3-(2-cyclopropyl-6-methylphenoxy)-4-pyridazinol (CAS 499223-49-3) and its salts and esters;
The further herbicidal compound B (component B) is preferably selected from the herbicides of class b1) to b15):
including their agriculturally acceptable salts or derivatives.
Preference is given to those compositions according to the present invention comprising at least one herbicide B selected from herbicides of class b2, b3, b4, b5, b6, b7, b9, b10 and b13.
Specific preference is given to those compositions according to the present invention which comprise at least one herbicide B selected from the herbicides of class b4, b6, b7, b9, b10 and b13.
Particular preference is given to those compositions according to the present invention which comprise at least one herbicide B selected from the herbicides of class b4, b6, b10 and b13.
Examples of herbicides B which can be used in combination with the uracilpyridines of formula (I) of formula (I) according to the present invention are:
b1) from the group of the lipid biosynthesis inhibitors:
ACC-herbicides such as alloxydim, alloxydim-sodium, butroxydim, clethodim, clodinafop, clodinafop-propargyl, cycloxydim, cyhalofop, cyhalofop-butyl, diclofop, diclofop-methyl, fenoxaprop, fenoxaprop-ethyl, fenoxaprop-P, fenoxaprop-P-ethyl, fluazifop, fluazifop-butyl, fluazifop-P, fluazifop-P-butyl, haloxyfop, haloxyfop-methyl, haloxyfop-P, haloxyfop-P-methyl, metamifop, pinoxaden, profoxydim, propaquizafop, quizalofop, quizalofop-ethyl, quizalofop-tefuryl, quizalofop-P, quizalofop-P-ethyl, quizalofop-P-tefuryl, sethoxydim, tepraloxydim, tralkoxydim,
4-(4′-Chloro-4-cyclopropyl-2′-fluoro[1,1′-biphenyl]-3-yl)-5-hydroxy-2,2,6,6-tetramethyl-2H-pyran-3 (6H)-one (CAS 1312337-72-6); 4-(2′,4′-Dichloro-4-cyclopropyl[1,1′-biphenyl]-3-yl)-5-hydroxy-2,2,6,6-tetramethyl-2H-pyran-3(6H)-one (CAS 1312337-45-3); 4-(4′-Chloro-4-ethyl-2′-fluoro[1,1′-biphenyl]-3-yl)-5-hydroxy-2,2,6,6-tetramethyl-2H-pyran-3(6H)-one (CAS 1033757-93-5); 4-(2′,4′-Dichloro-4-ethyl[1,1′-biphenyl]-3-yl)-2,2,6,6-tetramethyl-2H-pyran-3,5(4H,6H)-dione (CAS 1312340-84-3); 5-(Acetyloxy)-4-(4′-chloro-4-cyclopropyl-2′-fluoro[1,1′-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1312337-48-6); 5-(Acetyloxy)-4-(2′,4′-dichloro-4-cyclopropyl-[1,1′-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one; 5-(Acetyloxy)-4-(4′-chloro-4-ethyl-2′-fluoro[1,1′-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1312340-82-1); 5-(Acetyloxy)-4-(2′,4′-dichloro-4-ethyl[1,1′-biphenyl]-3-yl)-3,6-dihydro-2,2,6,6-tetramethyl-2H-pyran-3-one (CAS 1033760-55-2); 4-(4′-Chloro-4-cyclopropyl-2′-fluoro[1,1′-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester (CAS 1312337-51-1); 4-(2′,4′-Dichloro-4-cyclopropyl-[1,1′-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester; 4-(4′-Chloro-4-ethyl-2′-fluoro[1,1′-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester (CAS 1312340-83-2); 4-(2′,4′-Dichloro-4-ethyl[1,1′-biphenyl]-3-yl)-5,6-dihydro-2,2,6,6-tetramethyl-5-oxo-2H-pyran-3-yl carbonic acid methyl ester (CAS 1033760-58-5); and non ACC herbicides such as benfuresate, butylate, cycloate, dalapon, dimepiperate, EPTC, esprocarb, ethofumesate, flupropanate, molinate, orbencarb, pebulate, prosulfocarb, TCA, thiobencarb, tiocarbazil, triallate and vernolate;
b2) from the group of the ALS inhibitors:
sulfonylureas such as amidosulfuron, azimsulfuron, bensulfuron, bensulfuron-methyl, chlorimuron, chlorimuron-ethyl, chlorsulfuron, cinosulfuron, cyclosulfamuron, ethametsulfuron, ethametsulfuron-methyl, ethoxysulfuron, flazasulfuron, flucetosulfuron, flupyrsulfuron, flupyrsulfuron-methyl-sodium, foramsulfuron, halosulfuron, halosulfuron-methyl, imazosulfuron, iodosulfuron, iodosulfuron-methyl-sodium, iofensulfuron, iofensulfuron-sodium, mesosulfuron, metazosulfuron, metsulfuron, metsulfuron-methyl, nicosulfuron, orthosulfamuron, oxasulfuron, primisulfuron, primisulfuron-methyl, propyrisulfuron, prosulfuron, pyrazosulfuron, pyrazosulfuron-ethyl, rimsulfuron, sulfometuron, sulfometuron-methyl, sulfosulfuron, thifensulfuron, thifensulfuron-methyl, triasulfuron, tribenuron, tribenuron-methyl, trifloxysulfuron, triflusulfuron, triflusulfuron-methyl and tritosulfuron,
imidazolinones such as imazamethabenz, imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin and imazethapyr, triazolopyrimidine herbicides and sulfonanilides such as cloransulam, cloransulam-methyl, diclosulam, flumetsulam, florasulam, metosulam, penoxsulam, pyrimisulfan and pyroxsulam,
pyrimidinylbenzoates such as bispyribac, bispyribac-sodium, pyribenzoxim, pyriftalid, pyriminobac, pyriminobac-methyl, pyrithiobac, pyrithiobac-sodium, 4-[[[2-[(4,6-dimethoxy-2-pyrimidinyl)oxy]phenyl]methyl]amino]-benzoic acid-1-methylethyl ester (CAS 420138-41-6), 4-[[[2-[(4,6-dimethoxy-2-pyrimidinyl)oxy]phenyl]methyl]amino]-benzoic acid propyl ester (CAS 420138-40-5), N-(4-bromophenyl)-2-[(4,6-dimethoxy-2-pyrimidinyl)oxy]benzenemethanamine (CAS 420138-01-8),
sulfonylaminocarbonyl-triazolinone herbicides such as flucarbazone, flucarbazone-sodium, propoxycarbazone, propoxycarbazone-sodium, thiencarbazone and thiencarbazone-methyl; and triafamone;
among these, a preferred embodiment of the invention relates to those compositions comprising at least one imidazolinone herbicide;
b3) from the group of the photosynthesis inhibitors:
amicarbazone, inhibitors of the photosystem II, e.g. 1-(6-tert-butylpyrimidin-4-yl)-2-hydroxy-4-methoxy-3-methyl-2H-pyrrol-5-one (CAS 1654744-66-7), 1-(5-tert-butylisoxazol-3-yl)-2-hydroxy-4-methoxy-3-methyl-2H-pyrrol-5-one (CAS 1637455-12-9), 1-(5-tert-butylisoxazol-3-yl)-4-chloro-2-hydroxy-3-methyl-2H-pyrrol-5-one (CAS 1637453-94-1), 1-(5-tert-butyl-1-methyl-pyrazol-3-yl)-4-chloro-2-hydroxy-3-methyl-2H-pyrrol-5-one (CAS 1654057-29-0), 1-(5-tert-butyl-1-methyl-pyrazol-3-yl)-3-chloro-2-hydroxy-4-methyl-2H-pyrrol-5-one (CAS 1654747-80-4), 4-hydroxy-1-methoxy-5-methyl-3-[4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one; (CAS 2023785-78-4), 4-hydroxy-1,5-dimethyl-3-[4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one (CAS 2023785-79-5), 5-ethoxy-4-hydroxy-1-methyl-3-[4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one (CAS 1701416-69-4), 4-hydroxy-1-methyl-3-[4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one (CAS 1708087-22-2), 4-hydroxy-1,5-dimethyl-3-[1-methyl-5-(trifluoromethyl)pyrazol-3-yl]imidazolidin-2-one (CAS 2023785-80-8), 1-(5-tert-butylisoxazol-3-yl)-4-ethoxy-5-hydroxy-3-methyl-imidazolidin-2-one (CAS 1844836-64-1), triazine herbicides, including of chlorotriazine, triazinones, triazindiones, methylthiotriazines and pyridazinones such as ametryn, atrazine, chloridazone, cyanazine, desmetryn, dimethametryn,hexazinone, metribuzin, prometon, prometryn, propazine, simazine, simetryn, terbumeton, terbuthylazin, terbutryn and trietazin, aryl urea such as chlorobromuron, chlorotoluron, chloroxuron, dimefuron, diuron, fluometuron, isoproturon, isouron, linuron, metamitron, methabenzthiazuron, metobenzuron, metoxuron, monolinuron, neburon, siduron, tebuthiuron and thiadiazuron, phenyl carbamates such as desmedipham, karbutilat, phenmedipham, phenmedipham-ethyl, nitrile herbicides such as bromofenoxim, bromoxynil and its salts and esters, ioxynil and its salts and esters, uraciles such as bromacil, lenacil and terbacil, and bentazon and bentazon-sodium, pyridate, pyridafol, pentanochlor and propanil and inhibitors of the photosystem I such as diquat, diquat-dibromide, paraquat, paraquat-dichloride and paraquat-dimetilsulfate. Among these, a preferred embodiment of the invention relates to those compositions comprising at least one aryl urea herbicide. Among these, likewise a preferred embodiment of the invention relates to those compositions comprising at least one triazine herbicide. Among these, likewise a preferred embodiment of the invention relates to those compositions comprising at least one nitrile herbicide;
b4) from the group of the protoporphyrinogen-IX oxidase inhibitors:
acifluorfen, acifluorfen-sodium, azafenidin, bencarbazone, benzfendizone, bifenox, butafenacil, carfentrazone, carfentrazone-ethyl, chlomethoxyfen, chlorphtalim, cinidon-ethyl, fluazolate, flufenpyr, flufenpyr-ethyl, flumiclorac, flumiclorac-pentyl, flumioxazin, fluoroglycofen, fluoroglycofen-ethyl, fluthiacet, fluthiacet-methyl, fomesafen, halosafen, lactofen, oxadiargyl, oxadiazon, oxyfluorfen, pentoxazone, profluazol, pyraclonil, pyraflufen, pyraflufen-ethyl, saflufenacil, sulfentrazone, thidiazimin, tiafenacil, trifludimoxazin, ethyl [3-[2-chloro-4-fluoro-5-(1-methyl-6-trifluoromethyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-3-yl)phenoxy]-2-pyridyloxy]acetate (CAS 353292-31-6; S-3100, N-ethyl-3-(2,6-dichloro-4-trifluoro-methylphenoxy)-5-methyl-1H-pyrazole-1-carboxamide (CAS 452098-92-9), N-tetrahydrofurfuryl-3-(2,6-dichloro-4-trifluoromethylphenoxy)-5-methyl-1H-pyrazole-1-carboxamide (CAS 915396-43-9), N-ethyl-3-(2-chloro-6-fluoro-4-trifluoromethylphenoxy)-5-methyl-1H-pyrazole-1-carboxamide (CAS 452099-05-7), N-tetrahydrofurfuryl-3-(2-chloro-6-fluoro-4-trifluoro-methylphenoxy)-5-methyl-1H-pyrazole-1-carboxamide (CAS 452100-03-7), 3-[7-fluoro-3-oxo-4-(prop-2-ynyl)-3,4-dihydro-2H-benzo[1,4]oxazin-6-yl]-1,5-dimethyl-6-thioxo-[1,3,5]triazinan-2,4-dione (CAS 451484-50-7), 2-(2,2,7-trifluoro-3-oxo-4-prop-2-ynyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yl)-4,5,6,7-tetrahydro-isoindole-1,3-dione (CAS 1300118-96-0), 1-methyl-6-trifluoromethyl-3-(2,2,7-trifluoro-3-oxo-4-prop-2-ynyl-3,4-dihydro-2H-benzo[1,4]oxazin-6-yl)-1H-pyrimidine-2,4-dione (CAS 1304113-05-0), methyl (E)-4-[2-chloro-5-[4-chloro-5-(difluoromethoxy)-1H-methyl-pyrazol-3-yl]-4-fluoro-phenoxy]-3-methoxy-but-2-enoate (CAS 948893-00-3), and 3-[7-chloro-5-fluoro-2-(trifluoromethyl)-1H-benzimidazol-4-yl]-1-methyl-6-(trifluoromethyl)-1H-pyrimidine-2,4-dione (CAS 212754-02-4);
b5) from the group of the bleacher herbicides:
PDS inhibitors: beflubutamid, diflufenican, fluridone, flurochloridone, flurtamone, norflurazon, picolinafen, and 4-(3-trifluoromethylphenoxy)-2-(4-trifluoromethylphenyl)pyrimidine (CAS 180608-33-7), HPPD inhibitors: benzobicyclon, benzofenap, bicyclopyrone, clomazone, fenquinotrione, isoxaflutole, mesotrione, oxotrione (CAS 1486617-21-3), pyrasulfotole, pyrazolynate, pyrazoxyfen, sulcotrione, tefuryltrione, tembotrione, tolpyralate, topramezone, bleacher, unknown target: aclonifen, amitrole flumeturon and 2-chloro-3-methylsulfanyl-N-(1-methyltetrazol-5-yl)-4-(trifluoromethyl)benzamide (CAS 1361139-71-0), 2-(2,4-dichlorophenyl)methyl-4,4-dimethyl-3-isoxazolidone (CAS 81777-95-9) and 2-(2,5-dichlorophenyl)methyl-4,4-dimethyl-3-isoxazolidinone (CAS 81778-66-7);
b6) from the group of the EPSP synthase inhibitors:
glyphosate,glyphosate-isopropylammonium,glyposate-potassiumandglyphosate-trimesium (sulfosate);
b7) from the group of the glutamine synthase inhibitors:
bilanaphos (bialaphos), bilanaphos-sodium, glufosinate, glufosinate-P and glufosinate-ammonium;
b8) from the group of the DHP synthase inhibitors:
asulam;
b9) from the group of the mitosis inhibitors:
compounds of group K1: dinitroanilines such as benfluralin, butralin, dinitramine, ethalfluralin, fluchloralin, oryzalin, pendimethalin, prodiamine and trifluralin, phosphoramidates such as amiprophos, amiprophos-methyl, and butamiphos, benzoic acid herbicides such as chlorthal, chlorthal-dimethyl, pyridines such as dithiopyr and thiazopyr, benzamides such as propyzamide and tebutam; compounds of group K2: carbetamide, chlorpropham, flamprop, flamprop-isopropyl, flamprop-methyl, flamprop-M-isopropyl, flamprop-M-methyl and propham; among these, compounds of group K1, in particular dinitroanilines are preferred;
b10) from the group of the VLCFA inhibitors:
chloroacetamides such as acetochlor, alachlor, amidochlor, butachlor, dimethachlor, dimethenamid, dimethenamid-P, metazachlor, metolachlor, metolachlor-S, pethoxamid, pretilachlor, propachlor, propisochlor and thenylchlor, oxyacetanilides such as flufenacet and mefenacet, acetanilides such as diphenamid, naproanilide, napropamide and napropamide-M, tetrazolinones such fentrazamide, and other herbicides such as anilofos, cafenstrole, fenoxasulfone, ipfencarbazone, piperophos, pyroxasulfone and isoxazoline compounds of the formulae II.1, II.2, II.3, II.4, II.5, II.6, II.7, II.8 and II.9
the isoxazoline compounds of the formula (I)I are known in the art, e.g. from WO 2006/024820, WO 2006/037945, WO 2007/071900 and WO 2007/096576;
among the VLCFA inhibitors, preference is given to chloroacetamides and oxyacetamides;
b11) from the group of the cellulose biosynthesis inhibitors:
4
chlorthiamid, dichlobenil, flupoxam, indaziflam, isoxaben, triaziflam and 1-cyclohexyl-5-pentafluorphenyloxy-1-[1,2,4,6]thiatriazin-3-ylamine (CAS 175899-01-1);
b12) from the group of the decoupler herbicides:
dinoseb, dinoterb and DNOC and its salts;
b13) from the group of the auxinic herbicides:
2,4-D and its salts and esters such as clacyfos, 2,4-DB and its salts and esters, aminocyclopyrachlor and its salts and esters, aminopyralid and its salts such as aminopyralid-dimethylammonium, aminopyralid-tris(2-hydroxypropyl)ammonium and its esters, benazolin, benazolin-ethyl, chloramben and its salts and esters, clomeprop, clopyralid and its salts and esters, dicamba and its salts and esters, dichlorprop and its salts and esters, dichlorprop-P and its salts and esters, flopyrauxifen, fluroxypyr, fluroxypyr-butometyl, fluroxypyr-meptyl, halauxifen and its salts and esters (CAS 943832-60-8); MCPA and its salts and esters, MCPA-thioethyl, MCPB and its salts and esters, mecoprop and its salts and esters, mecoprop-P and its salts and esters, picloram and its salts and esters, quinclorac, quinmerac, TBA (2,3,6) and its salts and esters, triclopyr and its salts and esters, florpyrauxifen, florpyrauxifen-benzyl (CAS 1390661-72-9) and 4-amino-3-chloro-5-fluoro-6-(7-fluoro-1H-indol-6-yl)picolinic acid (CAS 1629965-65-6);
b14) from the group of the auxin transport inhibitors: diflufenzopyr, diflufenzopyr-sodium, naptalam and naptalam-sodium;
b15) from the group of the other herbicides: bromobutide, chlorflurenol, chlorflurenol-methyl, cinmethylin, cumyluron, cyclopyrimorate (CAS 499223-49-3) and its salts and esters, dalapon, dazomet, difenzoquat, difenzoquat-metilsulfate, dimethipin, DSMA, dymron, endothal and its salts, etobenzanid, flurenol, flurenol-butyl, flurprimidol, fosamine, fosamine-ammonium, indanofan, maleic hydrazide, mefluidide, metam, methiozolin (CAS 403640-27-7), methyl azide, methyl bromide, methyl-dymron, methyl iodide, MSMA, oleic acid, oxaziclomefone, pelargonic acid, pyributicarb, quinoclamine and tridiphane.
Active compounds B and C having a carboxyl group can be employed in the form of the acid, in the form of an agriculturally suitable salt as mentioned above or else in the form of an agriculturally acceptable derivative in the compositions according to the invention.
In the case of dicamba, suitable salts include those, where the counterion is an agriculturally acceptable cation. For example, suitable salts of dicamba are dicamba-sodium, dicamba-potassium, dicamba-methylammonium, dicamba-dimethylammonium, dicamba-isopropylammonium, dicamba-diglycolamine, dicamba-olamine, dicamba-diolamine, dicamba-trolamine, dicamba-N,N-bis-(3-aminopropyl)methylamine and dicamba-diethylenetriamine. Examples of a suitable ester are dicamba-methyl and dicamba-butotyl.
Suitable salts of 2,4-D are 2,4-D-ammonium, 2,4-D-dimethylammonium, 2,4-D-diethylammonium, 2,4-D-diethanolammonium (2,4-D-diolamine), 2,4-D-triethanolammonium, 2,4-D-isopropylammonium, 2,4-D-triisopropanolammonium, 2,4-D-heptylammonium, 2,4-D-dodecylammonium, 2,4-D-tetradecylammonium, 2,4-D-triethylammonium, 2,4-D-tris(2-hydroxypropyl)ammonium, 2,4-D-tris(isopropyl)ammonium, 2,4-D-trolamine, 2,4-D-lithium, 2,4-D-sodium. Examples of suitable esters of 2,4-D are 2,4-D-butotyl, 2,4-D-2-butoxypropyl, 2,4-D-3-butoxypropyl, 2,4-D-butyl, 2,4-D-ethyl, 2,4-D-ethylhexyl, 2,4-D-isobutyl, 2,4-D-isooctyl, 2,4-D-isopropyl, 2,4-D-meptyl, 2,4-D-methyl, 2,4-D-octyl, 2,4-D-pentyl, 2,4-D-propyl, 2,4-D-tefuryl and clacyfos.
Suitable salts of 2,4-DB are for example 2,4-DB-sodium, 2,4-DB-potassium and 2,4-DB-dimethylammonium. Suitable esters of 2,4-DB are for example 2,4-DB-butyl and 2,4-DB-isoctyl. Suitable salts of dichlorprop are for example dichlorprop-sodium, dichlorprop-potassium and dichlorprop-dimethylammonium. Examples of suitable esters of dichlorprop are dichlorprop-butotyl and dichlorprop-isoctyl.
Suitable salts and esters of MCPA include MCPA-butotyl, MCPA-butyl, MCPA-dimethylammonium, MCPA-diolamine, MCPA-ethyl, MCPA-thioethyl, MCPA-2-ethylhexyl, MCPA-isobutyl, MCPA-isoctyl, MCPA-isopropyl, MCPA-isopropylammonium, MCPA-methyl, MCPA-olamine, MCPA-potassium, MCPA-sodium and MCPA-trolamine.
A suitable salt of MCPB is MCPB sodium. A suitable ester of MCPB is MCPB-ethyl. Suitable salts of clopyralid are clopyralid-potassium, clopyralid-olamine and clopyralid-tris-(2-hydroxypropyl)ammonium. Example of suitable esters of clopyralid is clopyralid-methyl. Examples of a suitable ester of fluroxypyr are fluroxypyr-meptyl and fluroxypyr-2-butoxy-1-methylethyl, wherein fluroxypyr-meptyl is preferred.
Suitable salts of picloram are picloram-dimethylammonium, picloram-potassium, picloram-triisopropanolammonium, picloram-triisopropylammonium and picloram-trolamine. A suitable ester of picloram is picloram-isoctyl.
A suitable salt of triclopyr is triclopyr-triethylammonium. Suitable esters of triclopyr are for example triclopyr-ethyl and triclopyr-butotyl.
Suitable salts and esters of chloramben include chloramben-ammonium, chloramben-diolamine, chloramben-methyl, chloramben-methylammonium and chloramben-sodium. Suitable salts and esters of 2,3,6-TBA include 2,3,6-TBA-dimethylammonium, 2,3,6-TBA-lithium, 2,3,6-TBA-potassium and 2,3,6-TBA-sodium.
Suitable salts and esters of aminopyralid include aminopyralid-potassium, aminopyralid-dimethylammonium, and aminopyralid-tris(2-hydroxypropyl)ammonium.
Suitable salts of glyphosate are for example glyphosate-ammonium, glyphosate-diammonium, glyphoste-dimethylammonium, glyphosate-isopropylammonium, glyphosate-potassium, glyphosate-sodium, glyphosate-trimesium as well as the ethanolamine and diethanolamine salts, preferably glyphosate-diammonium, glyphosate-isopropylammonium and glyphosate-trimesium (sulfosate).
A suitable salt of glufosinate is for example glufosinate-ammonium.
A suitable salt of glufosinate-P is for example glufosinate-P-ammonium.
Suitable salts and esters of bromoxynil are for example bromoxynil-butyrate, bromoxynil-heptanoate, bromoxynil-octanoate, bromoxynil-potassium and bromoxynil-sodium.
Suitable salts and esters of ioxonil are for example ioxonil-octanoate, ioxonil-potassium and ioxonil-sodium.
Suitable salts and esters of mecoprop include mecoprop-butotyl, mecoprop-dimethylammonium, mecoprop-diolamine, mecoprop-ethadyl, mecoprop-2-ethylhexyl, mecoprop-isoctyl, mecoprop-methyl, mecoprop-potassium, mecoprop-sodium and mecoprop-trolamine.
Suitable salts of mecoprop-P are for example mecoprop-P-butotyl, mecoprop-P-dimethylammonium, mecoprop-P-2-ethylhexyl, mecoprop-P-isobutyl, mecoprop-P-potassium and mecoprop-P-sodium.
A suitable salt of diflufenzopyr is for example diflufenzopyr-sodium.
A suitable salt of naptalam is for example naptalam-sodium.
Suitable salts and esters of aminocyclopyrachlor are for example aminocyclopyrachlor-dimethylammonium, aminocyclopyrachlor-methyl, aminocyclopyrachlor-triisopropanolammonium, aminocyclopyrachlor-sodium and aminocyclopyrachlor-potassium.
A suitable salt of quinclorac is for example quinclorac-dimethylammonium.
A suitable salt of quinmerac is for example quinmerac-dimethylammonium.
A suitable salt of imazamox is for example imazamox-ammonium.
Suitable salts of imazapic are for example imazapic-ammonium and imazapic-isopropylammonium.
Suitable salts of imazapyr are for example imazapyr-ammonium and imazapyr-isopropylammonium.
A suitable salt of imazaquin is for example imazaquin-ammonium.
Suitable salts of imazethapyr are for example imazethapyr-ammonium and imazethapyr-isopropylammonium.
A suitable salt of topramezone is for example topramezone-sodium.
Particularly preferred herbicides Bare the herbicides Bas defined above; in particular the herbicides B.1-B.202, especially the herbicides B.1-B.201 listed below in table B:
TABLE B
Herbicide B
A. 1
clethodim
A. 2
clodinafop-propargyl
A. 3
cycloxydim
A. 4
cyhalofop-butyl
A. 5
fenoxaprop-ethyl
A. 6
fenoxaprop-P-ethyl
A. 7
metamifop
A. 8
pinoxaden
A. 9
profoxydim
A. 10
sethoxydim
A. 11
tepraloxydim
A. 12
tralkoxydim
A. 13
esprocarb
A. 14
ethofumesate
A. 15
molinate
A. 16
prosulfocarb
A. 17
thiobencarb
A. 18
triallate
A. 19
bensulfuron-methyl
A. 20
bispyribac-sodium
A. 21
cloransulam-methyl
A. 22
chlorsulfuron
A. 23
clorimuron
A. 24
cyclosulfamuron
A. 25
diclosulam
A. 26
florasulam
A. 27
flumetsulam
A. 28
flupyrsulfuron-methyl-
sodium
A. 29
foramsulfuron
A. 30
imazamox
A. 31
imazamox-ammonium
A. 32
imazapic
A. 33
imazapic-ammonium
A. 34
imazapic-
isopropylammonium
A. 35
imazapyr
A. 36
imazapyr-ammonium
A. 37
imazapyr-
isopropylammonium
A. 38
imazaquin
A. 39
imazaquin-ammonium
A. 40
imazethapyr
A. 41
imazethapyr-ammonium
A. 42
imazethapyr-
isopropylammonium
A. 43
imazosulfuron
A. 44
iodosulfuron-methyl-
sodium
A. 45
iofensulfuron
A. 46
iofensulfuron-sodium
A. 47
mesosulfuron-methyl
A. 48
metazosulfuron
A. 49
metsulfuron-methyl
A. 50
metosulam
A. 51
nicosulfuron
A. 52
penoxsulam
A. 53
propoxycarbazon-sodium
A. 54
pyrazosulfuron-ethyl
A. 55
pyribenzoxim
A. 56
pyriftalid
A. 57
pyroxsulam
A. 58
propyrisulfuron
A. 59
rimsulfuron
A. 60
sulfosulfuron
A. 61
thiencarbazone-methyl
A. 62
thifensulfuron-methyl
A. 63
tribenuron-methyl
A. 64
tritosulfuron
A. 65
triafamone
A. 66
ametryne
A. 67
atrazine
A. 68
bentazon
A. 69
bromoxynil
A. 70
bromoxynil-octanoate
A. 71
bromoxynil-heptanoate
A. 72
bromoxynil-potassium
A. 73
diuron
A. 74
fluometuron
A. 75
hexazinone
A. 76
isoproturon
A. 77
linuron
A. 78
metamitron
A. 79
metribuzin
A. 80
propanil
A. 81
simazin
A. 82
terbuthylazine
A. 83
terbutryn
A. 84
paraquat-dichloride
A. 85
acifluorfen
A. 86
butafenacil
A. 87
carfentrazone-ethyl
A. 88
flumioxazin
A. 89
fomesafen
A. 90
oxadiargyl
A. 91
oxyfluorfen
A. 92
pyraflufen
A. 93
pyraflufen-ethyl
A. 94
saflufenacil
A. 95
sulfentrazone
A. 96
trifludimoxazin
A. 97
ethyl [3-[2-chloro-4-fluoro-
5-(1-methyl-6-
trifluoromethyl-2,4-dioxo-
1,2,3,4-tetrahydropyri-
midin-3-yl)phenoxy]-2-
pyridyloxy]acetate (CAS
353292-31-6
A. 98
benzobicyclon
A. 99
bicyclopyrone
A. 100
clomazone
A. 101
diflufenican
A. 102
flurochloridone
A. 103
isoxaflutole
A. 104
mesotrione
A. 105
norflurazone
A. 106
picolinafen
A. 107
sulcotrione
A. 108
tefuryltrione
A. 109
tembotrione
A. 110
tolpyralate
A. 111
topramezone
A. 112
topramezone-sodium
A. 113
amitrole
A. 114
fluometuron
A. 115
fenquinotrione
A. 116
glyphosate
A. 117
glyphosate-ammonium
A. 118
glyphosate-
dimethylammonium
A. 119
glyphosate-
isopropylammonium
A. 120
glyphosate-trimesium
(sulfosate)
A. 121
glyphosate-potassium
A. 122
glufosinate
A. 123
glufosinate-ammonium
A. 124
glufosinate-P
A. 125
glufosinate-P-ammonium
A. 126
pendimethalin
A. 127
trifluralin
A. 128
acetochlor
A. 129
butachlor
A. 130
cafenstrole
A. 131
dimethenamid-P
A. 132
fentrazamide
A. 133
flufenacet
A. 134
mefenacet
A. 135
metazachlor
A. 136
metolachlor
A. 137
S-metolachlor
A. 138
pretilachlor
A. 139
fenoxasulfone
A. 140
indaziflam
A. 141
isoxaben
A. 142
triaziflam
A. 143
ipfencarbazone
A. 144
pyroxasulfone
A. 145
2,4-D
A. 146
2,4-D-isobutyl
A. 147
2,4-D-dimethylammonium
A. 148
2,4-D-N,N,N-
trimethylethanolammonium
A. 149
aminopyralid
A. 150
aminopyralid-methyl
A. 151
aminopyralid-dimethyl-
ammonium
A. 152
aminopyralid-tris(2-
hydroxypropyl)ammonium
A. 153
clopyralid
A. 154
clopyralid-methyl
A. 155
clopyralid-olamine
A. 156
dicamba
A. 157
dicamba-butotyl
A. 158
dicamba-diglycolamine
A. 159
dicamba-
dimethylammonium
A. 160
dicamba-diolamine
A. 161
dicamba-
isopropylammonium
A. 162
dicamba-potassium
A. 163
dicamba-sodium
A. 164
dicamba-trolamine
A. 165
dicamba-N,N-bis-(3-
aminopropyl)methylamine
A. 166
dicamba-
diethylenetriamine
A. 167
fluroxypyr
A. 168
fluroxypyr-meptyl
A. 169
halauxifen
A. 170
halauxifen-methyl
A. 171
MCPA
A. 172
MCPA-2-ethylhexyl
A. 173
MCPA-dimethylammonium
A. 174
quinclorac
A. 175
quinclorac-
dimethylammonium
A. 176
quinmerac
A. 177
quinmerac-
dimethylammonium
A. 178
florpyrauxifen
A. 179
florpyrauxifen-benzyl (CAS
1390661-72-9)
A. 180
aminocyclopyrachlor
A. 181
aminocyclopyrachlor-
potassium
A. 182
aminocyclopyrachlor-
methyl
A. 183
diflufenzopyr
A. 184
diflufenzopyr-sodium
A. 185
dymron
A. 186
indanofan
A. 187
oxaziclomefone
A. 188
II.1
A. 189
II.2
A. 190
II.3
A. 191
II.4
A. 192
II.5
A. 193
II.6
A. 194
II.7
A. 195
II.8
A. 196
II.9
A. 197
4-amino-3-chloro-5-fluoro-
6-(7-fluoro-1H-indol-6-
yl)picolinic acid (CAS
1629965-65-6)
A. 198
flopyrauxifen
A. 199
oxotrione (CAS 1486617-
21-3)
A. 200
cinmethylin
A. 201
2-chloro-3-methylsulfanyl-
N-(1-methyltetrazol-5-yl)-4-
(trifluoromethyl)benzamide
(CAS 1361139-71-0)
A. 202
2-(2,4-dichlorophenyl)-
methyl-4,4-dimethyl-3-
isoxazolidone (CAS
81777-95-9)
Moreover, it may be useful to apply the uracilpyridines of formula (I) in combination with safeners. Safeners are chemical compounds which prevent or reduce damage on useful plants without having a major impact on the herbicidal action of the uracilpyridines of formula (I) towards unwanted plants. They can be applied either before sowings (e.g. on seed treatments, shoots or seedlings) or in the pre-emergence application or post-emergence application of the useful plant. The safeners and the uracilpyridines of formula (I) and optionally the herbicides B can be applied simultaneously or in succession.
In another embodiment of the present invention the compositions according to the present invention comprise at least one uracilpyridine of formula (I) and at least one safener C (component C).
Suitable safeners are e.g. (quinolin-8-oxy)acetic acids, 1-phenyl-5-haloalkyl-1H-1,2,4-triazol-3-carboxylic acids, 1-phenyl-4,5-dihydro-5-alkyl-1H-pyrazol-3,5-dicarboxylic acids, 4,5-dihydro-5,5-diaryl-3-isoxazol carboxylic acids, dichloroacetamides, alpha-oximinophenylacetonitriles, acetophenonoximes, 4,6-dihalo-2-phenylpyrimidines, N-[[4-(aminocarbonyl)phenyl]sulfonyl]-2-benzoic amides, 1,8-naphthalic anhydride, 2-halo-4-(haloalkyl)-5-thiazol carboxylic acids, phosphorthiolates and N-alkyl-O-phenylcarbamates and their agriculturally acceptable salts and their agriculturally acceptable derivatives such amides, esters, and thioesters, provided they have an acid group.
Examples of preferred safeners C are benoxacor, cloquintocet, cyometrinil, cyprosulfamide, dichlormid, dicyclonon, dietholate, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, mephenate, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-1-oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3), 2,2,5-trimethyl-3-(dichloroacetyl)-1,3-oxazolidine (R-29148, CAS 52836-31-4), metcamifen and BPCMS (CAS 54091-06-4);
especially preferred benoxacor, cloquintocet, cyometrinil, cyprosulfamide, dichlormid, dicyclonon, dietholate, fenchlorazole, fenclorim, flurazole, fluxofenim, furilazole, isoxadifen, mefenpyr, mephenate, naphthalic anhydride, oxabetrinil, 4-(dichloroacetyl)-1-oxa-4-azaspiro[4.5]decane (MON4660, CAS 71526-07-3), 2,2,5-trimethyl-3-(dichloroacetyl)-1,3-oxazolidine (R-29148, CAS 52836-31-4) and metcamifen.
Particularly preferred safeners C, which, as component C, are constituent of the composition according to the invention are the safeners C as defined above; in particular the safeners C.1-C.17 listed below in table C:
TABLE C
Safener C
C. 1
benoxacor
C. 2
cloquintocet
C. 3
cloquintocet-mexyl
C. 4
cyprosulfamide
C. 5
dichlormid
C. 6
fenchlorazole
C. 7
fenchlorazole-ethyl
C. 8
fenclorim
C. 9
furilazole
C. 10
isoxadifen
C. 11
isoxadifen-ethyl
C. 12
mefenpyr
C. 13
mefenpyr-diethyl
C. 14
naphtalic acid anhydride
C. 15
4-(dichloroacetyl)-1-oxa-4-azaspiro[4.5]decane (CAS 71526-07-3)
C. 16
2,2,5-trimethyl-3-(dichloroacetyl)-1,3-oxazolidine (CAS
52836-31-4)
C. 17
metcamifen
The active compounds B of groups b1) to b15) and the active compounds C are known herbicides and safeners, see, for example, The Compendium of Pesticide Common Names (http://www.alanwood.net/pesticides/); Farm Chemicals Handbook 2000 volume 86, Meister Publishing Company, 2000; B. Hock, C. Fedtke, R. R. Schmidt, Herbizide [Herbicides], Georg Thieme Verlag, Stuttgart 1995; W. H. Ahrens, Herbicide Handbook, 7th edition, Weed Science Society of America, 1994; and K. K. Hatzios, Herbicide Handbook, Supplement for the 7th edition, Weed Science Society of America, 1998. 2,2,5-Trimethyl-3-(dichloroacetyl)-1,3-oxazolidine [CAS No. 52836-31-4] is also referred to as R-29148. 4-(Dichloroacetyl)-1-oxa-4-azaspiro[4.5]decane [CAS No. 71526-07-3] is also referred to as AD-67 and MON 4660.
The assignment of the active compounds to the respective mechanisms of action is based on current knowledge. If several mechanisms of action apply to one active compound, this substance was only assigned to one mechanism of action.
According to a preferred embodiment of the invention, the composition comprises as herbicidal active compound B or component B at least one, preferably exactly one herbicide B.
According to another preferred embodiment of the invention, the composition comprises as herbicidal active compounds B or component B at least two, preferably exactly two herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as herbicidal active compounds B or component B at least three, preferably exactly three herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as herbicidal active compounds B or component B at least four, preferably exactly four herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as safening component C or component C at least one, preferably exactly one safener C.
According to another preferred embodiment of the invention, the composition comprises as component B at least one, preferably exactly one herbicide B, and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises at least two, preferably exactly two, herbicides B different from each other, and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises at least three, preferably exactly three, herbicides B different from each other, and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), and as component B at least one, preferably exactly one, herbicide B.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), and at least two, preferably exactly two, herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), and at least three, preferably exactly three, herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), and at least four, preferably exactly four, herbicides B different from each other.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), as component B at least one, preferably exactly one, herbicide B, and as component C at least one, preferably exactly one safener C.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), at least two, preferably exactly two herbicides B different from each other, and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises as component A at least one, preferably exactly one compound of formula (I), preferably of formula (I.a) or (I.h), at least three, preferably exactly three herbicides B different from each other, and as component C at least one, preferably exactly one, safener C.
According to another preferred embodiment of the invention, the composition comprises, in addition to an uracilpyridine of formula (a), especially an active compound from the group consisting of (I.a.87), (I.a.109), (I.a.115), (I.a.255), (I.a.277), (I.a.283), (I.a.339), (I.a.361), (I.a.367), (I.h.87), (I.h.109), (I.h.115), (I.h.255), (I.h.277), (I.h.283), (I.h.339), (I.h.361) and (I.h.367),at least one and especially exactly one herbicidally active compound from group b4), in particular selected from the group consisting of acifluorfen, butafencil, carfenetrazone-ethyl, flumioxazin, fomesafen, oxadiargyl, oxyfluorfen, pyraflufen, pyraflufen-ethyl, saflufenacil, sulfentrazone, trifludimoxazin, ethyl [3-[2-chloro-4-fluoro-5-(1-methyl-6-trifluoromethyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-3-yl)phenoxy]-2-pyridloxy]acetate (CAS 353292-31-6). According to another preferred embodiment of the invention, the composition comprises, in addition to an uracilpyridine of formula (I), especially an active compound from the group consisting of (I.a.87), (I.a.109), (I.a.115), (I.a.255), (I.a.277), (I.a.283), (I.a.339), (I.a.361), (I.a.367), (I.h.87), (I.h.109), (I.h.115), (I.h.255), (I.h.277), (I.h.283), (I.h.339), (I.h.361) and (I.h.367), at least one and especially exactly one herbicidally active compound from group b6), in particular selected from the group consisting of glyphosate, glyphosate-ammonium, glyphosate-dimethylammonium, glyphosate-isopropylammonium and glyphosate-trimesium (sulfosate) and glyphosate-potassium.
According to another preferred embodiment of the invention, the composition comprises, in addition to an uracilpyridine of formula (I), especially an active compound from the group consisting of (I.a.87), (I.a.109), (I.a.115), (I.a.255), (I.a.277), (I.a.283), (I.a.339), (I.a.361), (I.a.367), (I.h.87), (I.h.109), (I.h.115), (I.h.255), (I.h.277), (I.h.283), (I.h.339), (I.h.361) and (I.h.367), at least one and especially exactly one herbicidally active compound from group b10), in particular selected from the group consisting of acetochlor, butachlor, cafenstrole, dimethenamid-P, fentrazamide, flufenacet, mefenacet, metazachlor, metolachlor, S-metolachlor, fenoxasulfone, ipfencarbazone and pyroxasulfone. Likewise, preference is given to compositions comprising in addition to an uracilpyridine of formula (I), especially an active compound from the group consisting of (I.a.87), (I.a.109), (I.a.115), (I.a.255), (I.a.277), (I.a.283), (I.a.339), (I.a.361), (I.a.367), (I.h.87), (I.h.109), (I.h.115), (I.h.255), (I.h.277), (I.h.283), (I.h.339), (I.h.361) and (I.h.367), at least one and especially exactly one herbicidally active compound from group b10), in particular selected from the group consisting of isoxazoline compounds of the formulae II.1, II.2, II.3, II.4, II.5, II.6, II.7, II.8 and II.9, as defined above.
According to another preferred embodiment of the invention, the composition comprises, in addition to an uracilpyridine of formula (I), especially an active compound from the group consisting of (I.a.87), (I.a.109), (I.a.115), (I.a.255), (I.a.277), (I.a.283), (I.a.339), (I.a.361), (I.a.367), (I.h.87), (I.h.109), (I.h.115), (I.h.255), (I.h.277), (I.h.283), (I.h.339), (I.h.361) and (I.h.367), at least one and especially exactly one herbicidally active compound from group b13), in particular selected from the group consisting of 2,4-D, 2,4-D-isobutyl, 2,4-D-dimethylammonium, 2,4-D-N,N,N-trimethylethanolammonium, aminocyclopyrachlor, aminocyclopyrachlor-potassium, aminocyclopyrachlor-methyl, aminopyralid, aminopyralid-methyl, aminopyralid-dimethylammonium, aminopyralid-tris(2-hydroxypropyl)ammonium, clopyralid, clopyralid-methyl, clopyralid-olamine, dicamba, dicamba-butotyl, dicamba-diglycolamine, dicamba-dimethylammonium, dicamba-diolamine, dicamba-isopropylammonium, dicamba-potassium, dicamba-sodium, dicamba-trolamine, dicamba-N,N-bis-(3-aminopropyl)methylamine, dicamba-diethylenetriamine, flopyrauxifen, fluroxypyr, fluroxypyr-meptyl, halauxifen, halauxifen-methyl, MCPA, MCPA-2-ethylhexyl, MCPA-dimethylammonium, quinclorac, quinclorac-dimethylammonium, quinmerac, quinmerac-dimethylammonium, florpyrauxifen, florpyrauxifen-benzyl (CAS 1390661-72-9), and 4-amino-3-chloro-5-fluoro-6-(7-fluoro-1H-indol-6-yl)picolinic acid.
Here and below, the term “binary compositions” includes compositions comprising one or more, for example 1, 2 or 3, active compounds of the formula (I) and either one or more, for example 1, 2 or 3, herbicides B or one or more safeners C.
Correspondingly, the term “ternary compositions” includes compositions comprising one or more, for example 1, 2 or 3, active compounds of the formula (I), one or more, for example 1, 2 or 3, herbicides B and one or more, for example 1, 2 or 3, safeners C.
In binary compositions comprising at least one compound of the formula (I) as component A and at least one herbicide B, the weight ratio of the active compounds A:B is generally in the range of from 1:1000 to 1000:1, preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:125 to 125:1.
In binary compositions comprising at least one compound of the formula (I) as component A and at least one safener C, the weight ratio of the active compounds A:C is generally in the range of from 1:1000 to 1000:1, preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:75 to 75:1.
In ternary compositions comprising at least one uracilpyridine of formula (I) as component A, at least one herbicide B and at least one safener C, the relative proportions by weight of the components A:B are generally in the range of from 1:1000 to 1000:1, preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:125 to 125:1, the weight ratio of the components A:C is generally in the range of from 1:1000 to 1000:1, preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:75 to 75:1, and the weight ratio of the components B:C is generally in the range of from 1:1000 to 1000:1, preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:75 to 75:1. The weight ratio of components A+B to component C is preferably in the range of from 1:500 to 500:1, in particular in the range of from 1:250 to 250:1 and particularly preferably in the range of from 1:75 to 75:1.
The weight ratios of the individual components in the preferred mixtures mentioned below are within the limits given herein, in particular within the preferred limits.
Particularly preferred are the compositions mentioned below comprising the uracilpyridines of formula (as defined and the substance(s) as defined in the respective row of table 1;
especially preferred comprising as only herbicidal active compounds the uracilpyridines of formula (I) as defined and the substance(s) as defined in the respective row of table 1;
most preferably comprising as only active compounds the uracilpyridines of formula (I) as defined and the substance(s) as defined in the respective row of table 1.
Particularly preferred are compositions 1.1 to 1.3653, especially compositions 1.1 to 1.3635, comprising the uracilpyridine (Ia.339) and the substance(s) as defined in the respective row of table 1:
TABLE 1
(compositions 1.1 to 1.3635):
comp.
no.
herbicide B
safener C
1.1
B.1
—
1.2
B.2
—
1.3
B.3
—
1.4
B.4
—
1.5
B.5
—
1.6
B.6
—
1.7
B.7
—
1.8
B.8
—
1.9
B.9
—
1.10
B.10
—
1.11
B.11
—
1.12
B.12
—
1.13
B.13
—
1.14
B.14
—
1.15
B.15
—
1.16
B.16
—
1.17
B.17
—
1.18
B.18
—
1.19
B.19
—
1.20
B.20
—
1.21
B.21
—
1.22
B.22
—
1.23
B.23
—
1.24
B.24
—
1.25
B.25
—
1.26
B.26
—
1.27
B.27
—
1.28
B.28
—
1.29
B.29
—
1.30
B.30
—
1.31
B.31
—
1.32
B.32
—
1.33
B.33
—
1.34
B.34
—
1.35
B.35
—
1.36
B.36
—
1.37
B.37
—
1.38
B.38
—
1.39
B.39
—
1.40
B.40
—
1.41
B.41
—
1.42
B.42
—
1.43
B.43
—
1.44
B.44
—
1.45
B.45
—
1.46
B.46
—
1.47
B.47
—
1.48
B.48
—
1.49
B.49
—
1.50
B.50
—
1.51
B.51
—
1.52
B.52
—
1.53
B.53
—
1.54
B.54
—
1.55
B.55
—
1.56
B.56
—
1.57
B.57
—
1.58
B.58.
—
1.59
B.59
—
1.60
B.60
—
1.61
B.61
—
1.62
B.62
—
1.63
B.63
—
1.64
B.64
—
1.65
B.65
—
1.66
B.66
—
1.67
B.67
—
1.68
B.68
—
1.69
B.69
—
1.70
B.70
—
1.71
B.71
—
1.72
B.72
—
1.73
B.73
—
1.74
B.74
—
1.75
B.75
—
1.76
B.76
—
1.77
B.77
—
1.78
B.78
—
1.79
B.79
—
1.80
B.80
—
1.81
B.81
—
1.82
B.82
—
1.83
B.83
—
1.84
B.84
—
1.85
B.85
—
1.86
B.86
—
1.87
B.87
—
1.88
B.88
—
1.89
B.89
—
1.90
B.90
—
1.91
B.91
—
1.92
B.92
—
1.93
B.93
—
1.94
B.94
—
1.95
B.95
—
1.96
B.96
—
1.97
B.97
—
1.98
B.98
—
1.99
B.99
—
1.100
B.100
—
1.101
B.101
—
1.102
B.102
—
1.103
B.103
—
1.104
B.104
—
1.105
B.105
—
1.106
B.106
—
1.107
B.107
—
1.108
B.108
—
1.109
B.109
—
1.110
B.110
—
1.111
B.111
—
1.112
B.112
—
1.113
B.113
—
1.114
B.114
—
1.115
B.115
—
1.116
B.116
—
1.117
B.117
—
1.118
B.118
—
1.119
B.119
—
1.120
B.120
—
1.121
B.121
—
1.122
B.122
—
1.123
B.123
—
1.124
B.124
—
1.125
B.125
—
1.126
B.126
—
1.127
B.127
—
1.128
B.128
—
1.129
B.129
—
1.130
B.130
—
1.131
B.131
—
1.132
B.132
—
1.133
B.133
—
1.134
B.134
—
1.135
B.135
—
1.136
B.136
—
1.137
B.137
—
1.138
B.138
—
1.139
B.139
—
1.140
B.140
—
1.141
B.141
—
1.142
B.142
—
1.143
B.143
—
1.144
B.144
—
1.145
B.145
—
1.146
B.146
—
1.147
B.147
—
1.148
B.148
—
1.149
B.149
—
1.150
B.150
—
1.151
B.151
—
1.152
B.152
—
1.153
B.153
—
1.154
B.154
—
1.155
B.155
—
1.156
B.156
—
1.157
B.157
—
1.158
B.158
—
1.159
B.159
—
1.160
B.160
—
1.161
B.161
—
1.162
B.162
—
1.163
B.163
—
1.164
B.164
—
1.165
B.165
—
1.166
B.166
—
1.167
B.167
—
1.168
B.168
—
1.169
B.169
—
1.170
B.170
—
1.171
B.171
—
1.172
B.172
—
1.173
B.173
—
1.174
B.174
—
1.175
B.175
—
1.176
B.176
—
1.177
B.177
—
1.178
B.178
—
1.179
B.179
—
1.180
B.180
—
1.181
B.181
—
1.182
B.182
—
1.183
B.183
—
1.184
B.184
—
1.185
B.185
—
1.186
B.186
—
1.187
B.187
—
1.188
B.188
—
1.189
B.189
—
1.190
B.190
—
1.191
B.191
—
1.192
B.192
—
1.193
B.193
—
1.194
B.194
—
1.195
B.195
—
1.196
B.196
—
1.197
B.197
—
1.198
B.198
—
1.199
B.199
—
1.200
B.200
—
1.201
B.201
—
1.202
B.1
C.1
1.203
B.2
C.1
1.204
B.3
C.1
1.205
B.4
C.1
1.206
B.5
C.1
1.207
B.6
C.1
1.208
B.7
C.1
1.209
B.8
C.1
1.210
B.9
C.1
1.211
B.10
C.1
1.212
B.11
C.1
1.213
B.12
C.1
1.214
B.13
C.1
1.215
B.14
C.1
1.216
B.15
C.1
1.217
B.16
C.1
1.218
B.17
C.1
1.219
B.18
C.1
1.220
B.19
C.1
1.221
B.20
C.1
1.222
B.21
C.1
1.223
B.22
C.1
1.224
B.23
C.1
1.225
B.24
C.1
1.226
B.25
C.1
1.227
B.26
C.1
1.228
B.27
C.1
1.229
B.28
C.1
1.230
B.29
C.1
1.231
B.30
C.1
1.232
B.31
C.1
1.233
B.32
C.1
1.234
B.33
C.1
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1.404
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1.405
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1.406
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1.407
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1.408
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1.409
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1.410
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1.411
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1.412
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1.414
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1.415
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1.416
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1.417
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1.418
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1.419
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1.420
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1.421
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1.425
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1.428
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1.432
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1.435
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1.436
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1.437
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1.438
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C.2
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1.463
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C.2
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C.2
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C.2
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C.2
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C.2
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C.2
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1.539
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1.543
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1.550
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1.555
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1.558
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1.563
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1.566
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1.567
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1.568
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1.569
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1.570
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1.571
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1.572
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1.573
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1.574
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1.575
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1.576
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1.577
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1.578
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1.579
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1.580
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1.581
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1.582
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1.583
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1.584
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1.589
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1.591
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1.593
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1.594
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1.595
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1.596
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1.599
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1.600
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1.603
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1.604
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1.605
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1.606
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1.607
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1.608
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1.609
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1.610
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1.611
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1.612
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1.614
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1.615
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1.616
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1.617
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1.618
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1.619
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1.620
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1.621
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1.622
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1.623
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1.624
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1.625
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1.626
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1.627
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1.628
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1.629
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1.630
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1.631
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1.632
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1.633
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1.634
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1.635
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1.637
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1.638
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1.639
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1.640
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1.641
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1.643
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1.644
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1.645
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1.646
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1.647
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1.648
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1.649
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1.650
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1.651
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1.652
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1.653
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1.654
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1.655
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1.656
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C.3
1.657
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C.3
1.658
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C.3
1.659
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C.3
1.660
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C.3
1.661
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C.3
1.662
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C.3
1.663
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C.3
1.664
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1.665
B.62
C.3
1.666
B.63
C.3
1.667
B.64
C.3
1.668
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C.3
1.669
B.66
C.3
1.670
B.67
C.3
1.671
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C.3
1.672
B.69
C.3
1.673
B.70
C.3
1.674
B.71
C.3
1.675
B.72
C.3
1.676
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C.3
1.677
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C.3
1.678
B.75
C.3
1.679
B.76
C.3
1.680
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C.3
1.681
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C.3
1.682
B.79
C.3
1.683
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C.3
1.684
B.81
C.3
1.685
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C.3
1.686
B.83
C.3
1.687
B.84
C.3
1.688
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C.3
1.689
B.86
C.3
1.690
B.87
C.3
1.691
B.88
C.3
1.692
B.89
C.3
1.693
B.90
C.3
1.694
B.91
C.3
1.695
B.92
C.3
1.696
B.93
C.3
1.697
B.94
C.3
1.698
B.95
C.3
1.699
B.96
C.3
1.700
B.97
C.3
1.701
B.98
C.3
1.702
B.99
C.3
1.703
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C.3
1.704
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C.3
1.705
B.102
C.3
1.706
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C.3
1.707
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C.3
1.708
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C.3
1.709
B.106
C.3
1.710
B.107
C.3
1.711
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C.3
1.712
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C.3
1.713
B.110
C.3
1.714
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C.3
1.715
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C.3
1.716
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C.3
1.717
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C.3
1.718
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1.719
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C.3
1.720
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C.3
1.721
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C.3
1.722
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C.3
1.723
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C.3
1.724
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C.3
1.725
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C.3
1.726
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C.3
1.727
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C.3
1.728
B.125
C.3
1.729
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C.3
1.730
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C.3
1.731
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C.3
1.732
B.129
C.3
1.733
B.130
C.3
1.734
B.131
C.3
1.735
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C.3
1.736
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C.3
1.737
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C.3
1.738
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1.739
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C.3
1.740
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C.3
1.741
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1.742
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C.3
1.743
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C.3
1.744
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C.3
1.745
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C.3
1.746
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1.747
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C.3
1.748
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1.749
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C.3
1.750
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C.3
1.751
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C.3
1.752
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1.753
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1.754
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1.755
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C.3
1.756
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1.757
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1.758
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1.759
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C.3
1.760
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1.761
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1.762
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1.763
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C.3
1.764
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C.3
1.765
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1.766
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1.767
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C.3
1.768
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C.3
1.769
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1.770
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1.771
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C.3
1.772
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C.3
1.773
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C.3
1.774
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C.3
1.775
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C.3
1.776
B.173
C.3
1.777
B.174
C.3
1.778
B.175
C.3
1.779
B.176
C.3
1.780
B.177
C.3
1.781
B.178
C.3
1.782
B.179
C.3
1.783
B.180
C.3
1.784
B.181
C.3
1.785
B.182
C.3
1.786
B.183
C.3
1.787
B.184
C.3
1.788
B.185
C.3
1.789
B.186
C.3
1.790
B.187
C.3
1.791
B.188
C.3
1.792
B.189
C.3
1.793
B.190
C.3
1.794
B.191
C.3
1.795
B.192
C.3
1.796
B.193
C.3
1.797
B.194
C.3
1.798
B.195
C.3
1.799
B.196
C.3
1.800
B.197
C.3
1.801
B.198
C.3
1.802
B.199
C.3
1.803
B.200
C.3
1.804
B.201
C.3
1.805
B.1
C.4
1.806
B.2
C.4
1.807
B.3
C.4
1.808
B.4
C.4
1.809
B.5
C.4
1.810
B.6
C.4
1.811
B.7
C.4
1.812
B.8
C.4
1.813
B.9
C.4
1.814
B.10
C.4
1.815
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C.4
1.816
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C.4
1.817
B.13
C.4
1.818
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C.4
1.819
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C.4
1.820
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C.4
1.821
B.17
C.4
1.822
B.18
C.4
1.823
B.19
C.4
1.824
B.20
C.4
1.825
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C.4
1.826
B.22
C.4
1.827
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C.4
1.828
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C.4
1.829
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C.4
1.830
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C.4
1.831
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C.4
1.832
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1.833
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C.4
1.834
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C.4
1.835
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C.4
1.836
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C.4
1.837
B.33
C.4
1.838
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C.4
1.839
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C.4
1.840
B.36
C.4
1.841
B.37
C.4
1.842
B.38
C.4
1.843
B.39
C.4
1.844
B.40
C.4
1.845
B.41
C.4
1.846
B.42
C.4
1.847
B.43
C.4
1.848
B.44
C.4
1.849
B.45
C.4
1.850
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C.4
1.851
B.47
C.4
1.852
B.48
C.4
1.853
B.49
C.4
1.854
B.50
C.4
1.855
B.51
C.4
1.856
B.52
C.4
1.857
B.53
C.4
1.858
B.54
C.4
1.859
B.55
C.4
1.860
B.56
C.4
1.861
B.57
C.4
1.862
B.58.
C.4
1.863
B.59
C.4
1.864
B.60
C.4
1.865
B.61
C.4
1.866
B.62
C.4
1.867
B.63
C.4
1.868
B.64
C.4
1.869
B.65
C.4
1.870
B.66
C.4
1.871
B.67
C.4
1.872
B.68
C.4
1.873
B.69
C.4
1.874
B.70
C.4
1.875
B.71
C.4
1.876
B.72
C.4
1.877
B.73
C.4
1.878
B.74
C.4
1.879
B.75
C.4
1.880
B.76
C.4
1.881
B.77
C.4
1.882
B.78
C.4
1.883
B.79
C.4
1.884
B.80
C.4
1.885
B.81
C.4
1.886
B.82
C.4
1.887
B.83
C.4
1.888
B.84
C.4
1.889
B.85
C.4
1.890
B.86
C.4
1.891
B.87
C.4
1.892
B.88
C.4
1.893
B.89
C.4
1.894
B.90
C.4
1.895
B.91
C.4
1.896
B.92
C.4
1.897
B.93
C.4
1.898
B.94
C.4
1.899
B.95
C.4
1.900
B.96
C.4
1.901
B.97
C.4
1.902
B.98
C.4
1.903
B.99
C.4
1.904
B.100
C.4
1.905
B.101
C.4
1.906
B.102
C.4
1.907
B.103
C.4
1.908
B.104
C.4
1.909
B.105
C.4
1.910
B.106
C.4
1.911
B.107
C.4
1.912
B.108
C.4
1.913
B.109
C.4
1.914
B.110
C.4
1.915
B.111
C.4
1.916
B.112
C.4
1.917
B.113
C.4
1.918
B.114
C.4
1.919
B.115
C.4
1.920
B.116
C.4
1.921
B.117
C.4
1.922
B.118
C.4
1.923
B.119
C.4
1.924
B.120
C.4
1.925
B.121
C.4
1.926
B.122
C.4
1.927
B.123
C.4
1.928
B.124
C.4
1.929
B.125
C.4
1.930
B.126
C.4
1.931
B.127
C.4
1.932
B.128
C.4
1.933
B.129
C.4
1.934
B.130
C.4
1.935
B.131
C.4
1.936
B.132
C.4
1.937
B.133
C.4
1.938
B.134
C.4
1.939
B.135
C.4
1.940
B.136
C.4
1.941
B.137
C.4
1.942
B.138
C.4
1.943
B.139
C.4
1.944
B.140
C.4
1.945
B.141
C.4
1.946
B.142
C.4
1.947
B.143
C.4
1.948
B.144
C.4
1.949
B.145
C.4
1.950
B.146
C.4
1.951
B.147
C.4
1.952
B.148
C.4
1.953
B.149
C.4
1.954
B.150
C.4
1.955
B.151
C.4
1.956
B.152
C.4
1.957
B.153
C.4
1.958
B.154
C.4
1.959
B.155
C.4
1.960
B.156
C.4
1.961
B.157
C.4
1.962
B.158
C.4
1.963
B.159
C.4
1.964
B.160
C.4
1.965
B.161
C.4
1.966
B.162
C.4
1.967
B.163
C.4
1.968
B.164
C.4
1.969
B.165
C.4
1.970
B.166
C.4
1.971
B.167
C.4
1.972
B.168
C.4
1.973
B.169
C.4
1.974
B.170
C.4
1.975
B.171
C.4
1.976
B.172
C.4
1.977
B.173
C.4
1.978
B.174
C.4
1.979
B.175
C.4
1.980
B.176
C.4
1.981
B.177
C.4
1.982
B.178
C.4
1.983
B.179
C.4
1.984
B.180
C.4
1.985
B.181
C.4
1.986
B.182
C.4
1.987
B.183
C.4
1.988
B.184
C.4
1.989
B.185
C.4
1.990
B.186
C.4
1.991
B.187
C.4
1.992
B.188
C.4
1.993
B.189
C.4
1.994
B.190
C.4
1.995
B.191
C.4
1.996
B.192
C.4
1.997
B.193
C.4
1.998
B.194
C.4
1.999
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C.4
1.1000
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C.4
1.1001
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C.4
1.1002
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1.1003
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C.4
1.1004
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C.4
1.1005
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1.1006
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1.1007
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1.1008
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1.1009
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1.1010
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1.1011
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1.1012
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1.1013
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1.1015
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1.1016
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1.1017
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1.1018
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1.1020
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1.1021
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1.1022
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1.1024
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1.1025
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1.1026
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1.1027
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1.1028
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1.1030
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1.1031
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1.1032
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1.1033
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1.1034
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1.1035
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1.1038
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1.1039
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1.1041
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1.1042
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1.1043
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1.1044
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1.1046
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1.1047
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1.1048
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1.1049
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1.1050
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1.1051
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1.1052
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1.1053
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1.1054
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1.1055
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1.1056
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C.5
1.1057
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C.5
1.1058
B.53
C.5
1.1059
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C.5
1.1060
B.55
C.5
1.1061
B.56
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1.1062
B.57
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1.1063
B.58.
C.5
1.1064
B.59
C.5
1.1065
B.60
C.5
1.1066
B.61
C.5
1.1067
B.62
C.5
1.1068
B.63
C.5
1.1069
B.64
C.5
1.1070
B.65
C.5
1.1071
B.66
C.5
1.1072
B.67
C.5
1.1073
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C.5
1.1074
B.69
C.5
1.1075
B.70
C.5
1.1076
B.71
C.5
1.1077
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C.5
1.1078
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C.5
1.1079
B.74
C.5
1.1080
B.75
C.5
1.1081
B.76
C.5
1.1082
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C.5
1.1083
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C.5
1.1084
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C.5
1.1085
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C.5
1.1086
B.81
C.5
1.1087
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C.5
1.1088
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C.5
1.1089
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C.5
1.1090
B.85
C.5
1.1091
B.86
C.5
1.1092
B.87
C.5
1.1093
B.88
C.5
1.1094
B.89
C.5
1.1095
B.90
C.5
1.1096
B.91
C.5
1.1097
B.92
C.5
1.1098
B.93
C.5
1.1099
B.94
C.5
1.1100
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C.5
1.1101
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C.5
1.1102
B.97
C.5
1.1103
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C.5
1.1104
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C.5
1.1105
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C.5
1.1106
B.101
C.5
1.1107
B.102
C.5
1.1108
B.103
C.5
1.1109
B.104
C.5
1.1110
B.105
C.5
1.1111
B.106
C.5
1.1112
B.107
C.5
1.1113
B.108
C.5
1.1114
B.109
C.5
1.1115
B.110
C.5
1.1116
B.111
C.5
1.1117
B.112
C.5
1.1118
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C.5
1.1119
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C.5
1.1120
B.115
C.5
1.1121
B.116
C.5
1.1122
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C.5
1.1123
B.118
C.5
1.1124
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C.5
1.1125
B.120
C.5
1.1126
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C.5
1.1127
B.122
C.5
1.1128
B.123
C.5
1.1129
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C.5
1.1130
B.125
C.5
1.1131
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C.5
1.1132
B.127
C.5
1.1133
B.128
C.5
1.1134
B.129
C.5
1.1135
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C.5
1.1136
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C.5
1.1137
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C.5
1.1138
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C.5
1.1139
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C.5
1.1140
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C.5
1.1141
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C.5
1.1142
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1.1143
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1.1144
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1.1145
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C.5
1.1146
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C.5
1.1147
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1.1148
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1.1149
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1.1150
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1.1151
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1.1152
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1.1153
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1.1154
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1.1155
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1.1156
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1.1157
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C.5
1.1158
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1.1159
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1.1160
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1.1161
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1.1162
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C.5
1.1163
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1.1164
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1.1165
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1.1166
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1.1167
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1.1168
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1.1169
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1.1170
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1.1171
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1.1172
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1.1173
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1.1174
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1.1175
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1.1176
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1.1177
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1.1178
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1.1179
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C.5
1.1180
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1.1181
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1.1182
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1.1183
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1.1184
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1.1185
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1.1186
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1.1187
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1.1188
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1.1189
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C.5
1.1190
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C.5
1.1191
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C.5
1.1192
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C.5
1.1193
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1.1194
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1.1195
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C.5
1.1196
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C.5
1.1197
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C.5
1.1198
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C.5
1.1199
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1.1200
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C.6
1.1260
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1.1261
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1.1262
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1.1263
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C.6
1.1265
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C.6
1.1266
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C.6
1.1267
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1.1268
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1.1269
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C.6
1.1270
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C.6
1.1271
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C.6
1.1272
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1.1273
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C.6
1.1277
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C.6
1.1298
B.92
C.6
1.1299
B.93
C.6
1.1300
B.94
C.6
1.1301
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C.6
1.1302
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C.6
1.1303
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C.6
1.1304
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C.6
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C.6
1.1306
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C.6
1.1307
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C.6
1.1308
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C.6
1.1309
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C.6
1.1310
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C.6
1.1311
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C.6
1.1312
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C.6
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C.6
1.1314
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C.6
1.1315
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C.6
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C.6
1.1317
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C.6
1.1318
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C.6
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C.6
1.1320
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C.6
1.1321
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C.6
1.1322
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C.6
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C.6
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C.6
1.1325
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C.6
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C.6
1.1327
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C.6
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C.6
1.1329
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C.6
1.1330
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C.6
1.1331
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C.6
1.1332
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C.6
1.1333
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C.6
1.1334
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C.6
1.1335
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C.6
1.1336
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C.6
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C.6
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1.1340
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1.1341
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1.1342
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1.1347
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1.1355
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1.1360
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1.1365
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1.1366
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1.1367
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1.1371
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1.1376
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1.1377
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1.1378
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1.1380
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C.6
1.1381
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C.6
1.1382
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1.1384
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1.1385
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1.1386
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C.6
1.1387
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1.1388
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C.6
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1.1395
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C.6
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C.6
1.1397
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C.6
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C.6
1.1399
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1.1400
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1.1401
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1.1450
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C.7
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1.1462
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C.7
1.1466
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C.7
1.1467
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C.7
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1.1470
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C.7
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C.7
1.1473
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1.1475
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1.1476
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C.7
1.1478
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C.7
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C.7
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C.7
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1.1485
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C.7
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C.7
1.1497
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C.7
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C.7
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C.7
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C.7
1.1501
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C.7
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C.7
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C.7
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C.7
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1.1539
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1.1662
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1.1663
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1.1664
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1.1665
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1.1666
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C.8
1.1667
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C.8
1.1668
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C.8
1.1669
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1.1670
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C.8
1.1671
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C.8
1.1672
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C.8
1.1673
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C.8
1.1674
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C.8
1.1675
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C.8
1.1676
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C.8
1.1677
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C.8
1.1678
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C.8
1.1679
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C.8
1.1680
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C.8
1.1681
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1.1682
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C.8
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C.8
1.1684
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C.8
1.1685
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1.1686
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1.1687
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C.8
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C.8
1.1697
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1.1809
B.201
C.8
1.1810
B.1
C.9
1.1811
B.2
C.9
1.1812
B.3
C.9
1.1813
B.4
C.9
1.1814
B.5
C.9
1.1815
B.6
C.9
1.1816
B.7
C.9
1.1817
B.8
C.9
1.1818
B.9
C.9
1.1819
B.10
C.9
1.1820
B.11
C.9
1.1821
B.12
C.9
1.1822
B.13
C.9
1.1823
B.14
C.9
1.1824
B.15
C.9
1.1825
B.16
C.9
1.1826
B.17
C.9
1.1827
B.18
C.9
1.1828
B.19
C.9
1.1829
B.20
C.9
1.1830
B.21
C.9
1.1831
B.22
C.9
1.1832
B.23
C.9
1.1833
B.24
C.9
1.1834
B.25
C.9
1.1835
B.26
C.9
1.1836
B.27
C.9
1.1837
B.28
C.9
1.1838
B.29
C.9
1.1839
B.30
C.9
1.1840
B.31
C.9
1.1841
B.32
C.9
1.1842
B.33
C.9
1.1843
B.34
C.9
1.1844
B.35
C.9
1.1845
B.36
C.9
1.1846
B.37
C.9
1.1847
B.38
C.9
1.1848
B.39
C.9
1.1849
B.40
C.9
1.1850
B.41
C.9
1.1851
B.42
C.9
1.1852
B.43
C.9
1.1853
B.44
C.9
1.1854
B.45
C.9
1.1855
B.46
C.9
1.1856
B.47
C.9
1.1857
B.48
C.9
1.1858
B.49
C.9
1.1859
B.50
C.9
1.1860
B.51
C.9
1.1861
B.52
C.9
1.1862
B.53
C.9
1.1863
B.54
C.9
1.1864
B.55
C.9
1.1865
B.56
C.9
1.1866
B.57
C.9
1.1867
B.58.
C.9
1.1868
B.59
C.9
1.1869
B.60
C.9
1.1870
B.61
C.9
1.1871
B.62
C.9
1.1872
B.63
C.9
1.1873
B.64
C.9
1.1874
B.65
C.9
1.1875
B.66
C.9
1.1876
B.67
C.9
1.1877
B.68
C.9
1.1878
B.69
C.9
1.1879
B.70
C.9
1.1880
B.71
C.9
1.1881
B.72
C.9
1.1882
B.73
C.9
1.1883
B.74
C.9
1.1884
B.75
C.9
1.1885
B.76
C.9
1.1886
B.77
C.9
1.1887
B.78
C.9
1.1888
B.79
C.9
1.1889
B.80
C.9
1.1890
B.81
C.9
1.1891
B.82
C.9
1.1892
B.83
C.9
1.1893
B.84
C.9
1.1894
B.85
C.9
1.1895
B.86
C.9
1.1896
B.87
C.9
1.1897
B.88
C.9
1.1898
B.89
C.9
1.1899
B.90
C.9
1.1900
B.91
C.9
1.1901
B.92
C.9
1.1902
B.93
C.9
1.1903
B.94
C.9
1.1904
B.95
C.9
1.1905
B.96
C.9
1.1906
B.97
C.9
1.1907
B.98
C.9
1.1908
B.99
C.9
1.1909
B.100
C.9
1.1910
B.101
C.9
1.1911
B.102
C.9
1.1912
B.103
C.9
1.1913
B.104
C.9
1.1914
B.105
C.9
1.1915
B.106
C.9
1.1916
B.107
C.9
1.1917
B.108
C.9
1.1918
B.109
C.9
1.1919
B.110
C.9
1.1920
B.111
C.9
1.1921
B.112
C.9
1.1922
B.113
C.9
1.1923
B.114
C.9
1.1924
B.115
C.9
1.1925
B.116
C.9
1.1926
B.117
C.9
1.1927
B.118
C.9
1.1928
B.119
C.9
1.1929
B.120
C.9
1.1930
B.121
C.9
1.1931
B.122
C.9
1.1932
B.123
C.9
1.1933
B.124
C.9
1.1934
B.125
C.9
1.1935
B.126
C.9
1.1936
B.127
C.9
1.1937
B.128
C.9
1.1938
B.129
C.9
1.1939
B.130
C.9
1.1940
B.131
C.9
1.1941
B.132
C.9
1.1942
B.133
C.9
1.1943
B.134
C.9
1.1944
B.135
C.9
1.1945
B.136
C.9
1.1946
B.137
C.9
1.1947
B.138
C.9
1.1948
B.139
C.9
1.1949
B.140
C.9
1.1950
B.141
C.9
1.1951
B.142
C.9
1.1952
B.143
C.9
1.1953
B.144
C.9
1.1954
B.145
C.9
1.1955
B.146
C.9
1.1956
B.147
C.9
1.1957
B.148
C.9
1.1958
B.149
C.9
1.1959
B.150
C.9
1.1960
B.151
C.9
1.1961
B.152
C.9
1.1962
B.153
C.9
1.1963
B.154
C.9
1.1964
B.155
C.9
1.1965
B.156
C.9
1.1966
B.157
C.9
1.1967
B.158
C.9
1.1968
B.159
C.9
1.1969
B.160
C.9
1.1970
B.161
C.9
1.1971
B.162
C.9
1.1972
B.163
C.9
1.1973
B.164
C.9
1.1974
B.165
C.9
1.1975
B.166
C.9
1.1976
B.167
C.9
1.1977
B.168
C.9
1.1978
B.169
C.9
1.1979
B.170
C.9
1.1980
B.171
C.9
1.1981
B.172
C.9
1.1982
B.173
C.9
1.1983
B.174
C.9
1.1984
B.175
C.9
1.1985
B.176
C.9
1.1986
B.177
C.9
1.1987
B.178
C.9
1.1988
B.179
C.9
1.1989
B.180
C.9
1.1990
B.181
C.9
1.1991
B.182
C.9
1.1992
B.183
C.9
1.1993
B.184
C.9
1.1994
B.185
C.9
1.1995
B.186
C.9
1.1996
B.187
C.9
1.1997
B.188
C.9
1.1998
B.189
C.9
1.1999
B.190
C.9
1.2000
B.191
C.9
1.2001
B.192
C.9
1.2002
B.193
C.9
1.2003
B.194
C.9
1.2004
B.195
C.9
1.2005
B.196
C.9
1.2006
B.197
C.9
1.2007
B.198
C.9
1.2008
B.199
C.9
1.2009
B.200
C.9
1.2010
B.201
C.9
1.2011
B.1
C.10
1.2012
B.2
C.10
1.2013
B.3
C.10
1.2014
B.4
C.10
1.2015
B.5
C.10
1.2016
B.6
C.10
1.2017
B.7
C.10
1.2018
B.8
C.10
1.2019
B.9
C.10
1.2020
B.10
C.10
1.2021
B.11
C.10
1.2022
B.12
C.10
1.2023
B.13
C.10
1.2024
B.14
C.10
1.2025
B.15
C.10
1.2026
B.16
C.10
1.2027
B.17
C.10
1.2028
B.18
C.10
1.2029
B.19
C.10
1.2030
B.20
C.10
1.2031
B.21
C.10
1.2032
B.22
C.10
1.2033
B.23
C.10
1.2034
B.24
C.10
1.2035
B.25
C.10
1.2036
B.26
C.10
1.2037
B.27
C.10
1.2038
B.28
C.10
1.2039
B.29
C.10
1.2040
B.30
C.10
1.2041
B.31
C.10
1.2042
B.32
C.10
1.2043
B.33
C.10
1.2044
B.34
C.10
1.2045
B.35
C.10
1.2046
B.36
C.10
1.2047
B.37
C.10
1.2048
B.38
C.10
1.2049
B.39
C.10
1.2050
B.40
C.10
1.2051
B.41
C.10
1.2052
B.42
C.10
1.2053
B.43
C.10
1.2054
B.44
C.10
1.2055
B.45
C.10
1.2056
B.46
C.10
1.2057
B.47
C.10
1.2058
B.48
C.10
1.2059
B.49
C.10
1.2060
B.50
C.10
1.2061
B.51
C.10
1.2062
B.52
C.10
1.2063
B.53
C.10
1.2064
B.54
C.10
1.2065
B.55
C.10
1.2066
B.56
C.10
1.2067
B.57
C.10
1.2068
B.58.
C.10
1.2069
B.59
C.10
1.2070
B.60
C.10
1.2071
B.61
C.10
1.2072
B.62
C.10
1.2073
B.63
C.10
1.2074
B.64
C.10
1.2075
B.65
C.10
1.2076
B.66
C.10
1.2077
B.67
C.10
1.2078
B.68
C.10
1.2079
B.69
C.10
1.2080
B.70
C.10
1.2081
B.71
C.10
1.2082
B.72
C.10
1.2083
B.73
C.10
1.2084
B.74
C.10
1.2085
B.75
C.10
1.2086
B.76
C.10
1.2087
B.77
C.10
1.2088
B.78
C.10
1.2089
B.79
C.10
1.2090
B.80
C.10
1.2091
B.81
C.10
1.2092
B.82
C.10
1.2093
B.83
C.10
1.2094
B.84
C.10
1.2095
B.85
C.10
1.2096
B.86
C.10
1.2097
B.87
C.10
1.2098
B.88
C.10
1.2099
B.89
C.10
1.2100
B.90
C.10
1.2101
B.91
C.10
1.2102
B.92
C.10
1.2103
B.93
C.10
1.2104
B.94
C.10
1.2105
B.95
C.10
1.2106
B.96
C.10
1.2107
B.97
C.10
1.2108
B.98
C.10
1.2109
B.99
C.10
1.2110
B.100
C.10
1.2111
B.101
C.10
1.2112
B.102
C.10
1.2113
B.103
C.10
1.2114
B.104
C.10
1.2115
B.105
C.10
1.2116
B.106
C.10
1.2117
B.107
C.10
1.2118
B.108
C.10
1.2119
B.109
C.10
1.2120
B.110
C.10
1.2121
B.111
C.10
1.2122
B.112
C.10
1.2123
B.113
C.10
1.2124
B.114
C.10
1.2125
B.115
C.10
1.2126
B.116
C.10
1.2127
B.117
C.10
1.2128
B.118
C.10
1.2129
B.119
C.10
1.2130
B.120
C.10
1.2131
B.121
C.10
1.2132
B.122
C.10
1.2133
B.123
C.10
1.2134
B.124
C.10
1.2135
B.125
C.10
1.2136
B.126
C.10
1.2137
B.127
C.10
1.2138
B.128
C.10
1.2139
B.129
C.10
1.2140
B.130
C.10
1.2141
B.131
C.10
1.2142
B.132
C.10
1.2143
B.133
C.10
1.2144
B.134
C.10
1.2145
B.135
C.10
1.2146
B.136
C.10
1.2147
B.137
C.10
1.2148
B.138
C.10
1.2149
B.139
C.10
1.2150
B.140
C.10
1.2151
B.141
C.10
1.2152
B.142
C.10
1.2153
B.143
C.10
1.2154
B.144
C.10
1.2155
B.145
C.10
1.2156
B.146
C.10
1.2157
B.147
C.10
1.2158
B.148
C.10
1.2159
B.149
C.10
1.2160
B.150
C.10
1.2161
B.151
C.10
1.2162
B.152
C.10
1.2163
B.153
C.10
1.2164
B.154
C.10
1.2165
B.155
C.10
1.2166
B.156
C.10
1.2167
B.157
C.10
1.2168
B.158
C.10
1.2169
B.159
C.10
1.2170
B.160
C.10
1.2171
B.161
C.10
1.2172
B.162
C.10
1.2173
B.163
C.10
1.2174
B.164
C.10
1.2175
B.165
C.10
1.2176
B.166
C.10
1.2177
B.167
C.10
1.2178
B.168
C.10
1.2179
B.169
C.10
1.2180
B.170
C.10
1.2181
B.171
C.10
1.2182
B.172
C.10
1.2183
B.173
C.10
1.2184
B.174
C.10
1.2185
B.175
C.10
1.2186
B.176
C.10
1.2187
B.177
C.10
1.2188
B.178
C.10
1.2189
B.179
C.10
1.2190
B.180
C.10
1.2191
B.181
C.10
1.2192
B.182
C.10
1.2193
B.183
C.10
1.2194
B.184
C.10
1.2195
B.185
C.10
1.2196
B.186
C.10
1.2197
B.187
C.10
1.2198
B.188
C.10
1.2199
B.189
C.10
1.2200
B.190
C.10
1.2201
B.191
C.10
1.2202
B.192
C.10
1.2203
B.193
C.10
1.2204
B.194
C.10
1.2205
B.195
C.10
1.2206
B.196
C.10
1.2207
B.197
C.10
1.2208
B.198
C.10
1.2209
B.199
C.10
1.2210
B.200
C.10
1.2211
B.201
C.10
1.2212
B.1
C.11
1.2213
B.2
C.11
1.2214
B.3
C.11
1.2215
B.4
C.11
1.2216
B.5
C.11
1.2217
B.6
C.11
1.2218
B.7
C.11
1.2219
B.8
C.11
1.2220
B.9
C.11
1.2221
B.10
C.11
1.2222
B.11
C.11
1.2223
B.12
C.11
1.2224
B.13
C.11
1.2225
B.14
C.11
1.2226
B.15
C.11
1.2227
B.16
C.11
1.2228
B.17
C.11
1.2229
B.18
C.11
1.2230
B.19
C.11
1.2231
B.20
C.11
1.2232
B.21
C.11
1.2233
B.22
C.11
1.2234
B.23
C.11
1.2235
B.24
C.11
1.2236
B.25
C.11
1.2237
B.26
C.11
1.2238
B.27
C.11
1.2239
B.28
C.11
1.2240
B.29
C.11
1.2241
B.30
C.11
1.2242
B.31
C.11
1.2243
B.32
C.11
1.2244
B.33
C.11
1.2245
B.34
C.11
1.2246
B.35
C.11
1.2247
B.36
C.11
1.2248
B.37
C.11
1.2249
B.38
C.11
1.2250
B.39
C.11
1.2251
B.40
C.11
1.2252
B.41
C.11
1.2253
B.42
C.11
1.2254
B.43
C.11
1.2255
B.44
C.11
1.2256
B.45
C.11
1.2257
B.46
C.11
1.2258
B.47
C.11
1.2259
B.48
C.11
1.2260
B.49
C.11
1.2261
B.50
C.11
1.2262
B.51
C.11
1.2263
B.52
C.11
1.2264
B.53
C.11
1.2265
B.54
C.11
1.2266
B.55
C.11
1.2267
B.56
C.11
1.2268
B.57
C.11
1.2269
B.58.
C.11
1.2270
B.59
C.11
1.2271
B.60
C.11
1.2272
B.61
C.11
1.2273
B.62
C.11
1.2274
B.63
C.11
1.2275
B.64
C.11
1.2276
B.65
C.11
1.2277
B.66
C.11
1.2278
B.67
C.11
1.2279
B.68
C.11
1.2280
B.69
C.11
1.2281
B.70
C.11
1.2282
B.71
C.11
1.2283
B.72
C.11
1.2284
B.73
C.11
1.2285
B.74
C.11
1.2286
B.75
C.11
1.2287
B.76
C.11
1.2288
B.77
C.11
1.2289
B.78
C.11
1.2290
B.79
C.11
1.2291
B.80
C.11
1.2292
B.81
C.11
1.2293
B.82
C.11
1.2294
B.83
C.11
1.2295
B.84
C.11
1.2296
B.85
C.11
1.2297
B.86
C.11
1.2298
B.87
C.11
1.2299
B.88
C.11
1.2300
B.89
C.11
1.2301
B.90
C.11
1.2302
B.91
C.11
1.2303
B.92
C.11
1.2304
B.93
C.11
1.2305
B.94
C.11
1.2306
B.95
C.11
1.2307
B.96
C.11
1.2308
B.97
C.11
1.2309
B.98
C.11
1.2310
B.99
C.11
1.2311
B.100
C.11
1.2312
B.101
C.11
1.2313
B.102
C.11
1.2314
B.103
C.11
1.2315
B.104
C.11
1.2316
B.105
C.11
1.2317
B.106
C.11
1.2318
B.107
C.11
1.2319
B.108
C.11
1.2320
B.109
C.11
1.2321
B.110
C.11
1.2322
B.111
C.11
1.2323
B.112
C.11
1.2324
B.113
C.11
1.2325
B.114
C.11
1.2326
B.115
C.11
1.2327
B.116
C.11
1.2328
B.117
C.11
1.2329
B.118
C.11
1.2330
B.119
C.11
1.2331
B.120
C.11
1.2332
B.121
C.11
1.2333
B.122
C.11
1.2334
B.123
C.11
1.2335
B.124
C.11
1.2336
B.125
C.11
1.2337
B.126
C.11
1.2338
B.127
C.11
1.2339
B.128
C.11
1.2340
B.129
C.11
1.2341
B.130
C.11
1.2342
B.131
C.11
1.2343
B.132
C.11
1.2344
B.133
C.11
1.2345
B.134
C.11
1.2346
B.135
C.11
1.2347
B.136
C.11
1.2348
B.137
C.11
1.2349
B.138
C.11
1.2350
B.139
C.11
1.2351
B.140
C.11
1.2352
B.141
C.11
1.2353
B.142
C.11
1.2354
B.143
C.11
1.2355
B.144
C.11
1.2356
B.145
C.11
1.2357
B.146
C.11
1.2358
B.147
C.11
1.2359
B.148
C.11
1.2360
B.149
C.11
1.2361
B.150
C.11
1.2362
B.151
C.11
1.2363
B.152
C.11
1.2364
B.153
C.11
1.2365
B.154
C.11
1.2366
B.155
C.11
1.2367
B.156
C.11
1.2368
B.157
C.11
1.2369
B.158
C.11
1.2370
B.159
C.11
1.2371
B.160
C.11
1.2372
B.161
C.11
1.2373
B.162
C.11
1.2374
B.163
C.11
1.2375
B.164
C.11
1.2376
B.165
C.11
1.2377
B.166
C.11
1.2378
B.167
C.11
1.2379
B.168
C.11
1.2380
B.169
C.11
1.2381
B.170
C.11
1.2382
B.171
C.11
1.2383
B.172
C.11
1.2384
B.173
C.11
1.2385
B.174
C.11
1.2386
B.175
C.11
1.2387
B.176
C.11
1.2388
B.177
C.11
1.2389
B.178
C.11
1.2390
B.179
C.11
1.2391
B.180
C.11
1.2392
B.181
C.11
1.2393
B.182
C.11
1.2394
B.183
C.11
1.2395
B.184
C.11
1.2396
B.185
C.11
1.2397
B.186
C.11
1.2398
B.187
C.11
1.2399
B.188
C.11
1.2400
B.189
C.11
1.2401
B.190
C.11
1.2402
B.191
C.11
1.2403
B.192
C.11
1.2404
B.193
C.11
1.2405
B.194
C.11
1.2406
B.195
C.11
1.2407
B.196
C.11
1.2408
B.197
C.11
1.2409
B.198
C.11
1.2410
B.199
C.11
1.2411
B.200
C.11
1.2412
B.201
C.11
1.2413
B.1
C.12
1.2414
B.2
C.12
1.2415
B.3
C.12
1.2416
B.4
C.12
1.2417
B.5
C.12
1.2418
B.6
C.12
1.2419
B.7
C.12
1.2420
B.8
C.12
1.2421
B.9
C.12
1.2422
B.10
C.12
1.2423
B.11
C.12
1.2424
B.12
C.12
1.2425
B.13
C.12
1.2426
B.14
C.12
1.2427
B.15
C.12
1.2428
B.16
C.12
1.2429
B.17
C.12
1.2430
B.18
C.12
1.2431
B.19
C.12
1.2432
B.20
C.12
1.2433
B.21
C.12
1.2434
B.22
C.12
1.2435
B.23
C.12
1.2436
B.24
C.12
1.2437
B.25
C.12
1.2438
B.26
C.12
1.2439
B.27
C.12
1.2440
B.28
C.12
1.2441
B.29
C.12
1.2442
B.30
C.12
1.2443
B.31
C.12
1.2444
B.32
C.12
1.2445
B.33
C.12
1.2446
B.34
C.12
1.2447
B.35
C.12
1.2448
B.36
C.12
1.2449
B.37
C.12
1.2450
B.38
C.12
1.2451
B.39
C.12
1.2452
B.40
C.12
1.2453
B.41
C.12
1.2454
B.42
C.12
1.2455
B.43
C.12
1.2456
B.44
C.12
1.2457
B.45
C.12
1.2458
B.46
C.12
1.2459
B.47
C.12
1.2460
B.48
C.12
1.2461
B.49
C.12
1.2462
B.50
C.12
1.2463
B.51
C.12
1.2464
B.52
C.12
1.2465
B.53
C.12
1.2466
B.54
C.12
1.2467
B.55
C.12
1.2468
B.56
C.12
1.2469
B.57
C.12
1.2470
B.58.
C.12
1.2471
B.59
C.12
1.2472
B.60
C.12
1.2473
B.61
C.12
1.2474
B.62
C.12
1.2475
B.63
C.12
1.2476
B.64
C.12
1.2477
B.65
C.12
1.2478
B.66
C.12
1.2479
B.67
C.12
1.2480
B.68
C.12
1.2481
B.69
C.12
1.2482
B.70
C.12
1.2483
B.71
C.12
1.2484
B.72
C.12
1.2485
B.73
C.12
1.2486
B.74
C.12
1.2487
B.75
C.12
1.2488
B.76
C.12
1.2489
B.77
C.12
1.2490
B.78
C.12
1.2491
B.79
C.12
1.2492
B.80
C.12
1.2493
B.81
C.12
1.2494
B.82
C.12
1.2495
B.83
C.12
1.2496
B.84
C.12
1.2497
B.85
C.12
1.2498
B.86
C.12
1.2499
B.87
C.12
1.2500
B.88
C.12
1.2501
B.89
C.12
1.2502
B.90
C.12
1.2503
B.91
C.12
1.2504
B.92
C.12
1.2505
B.93
C.12
1.2506
B.94
C.12
1.2507
B.95
C.12
1.2508
B.96
C.12
1.2509
B.97
C.12
1.2510
B.98
C.12
1.2511
B.99
C.12
1.2512
B.100
C.12
1.2513
B.101
C.12
1.2514
B.102
C.12
1.2515
B.103
C.12
1.2516
B.104
C.12
1.2517
B.105
C.12
1.2518
B.106
C.12
1.2519
B.107
C.12
1.2520
B.108
C.12
1.2521
B.109
C.12
1.2522
B.110
C.12
1.2523
B.111
C.12
1.2524
B.112
C.12
1.2525
B.113
C.12
1.2526
B.114
C.12
1.2527
B.115
C.12
1.2528
B.116
C.12
1.2529
B.117
C.12
1.2530
B.118
C.12
1.2531
B.119
C.12
1.2532
B.120
C.12
1.2533
B.121
C.12
1.2534
B.122
C.12
1.2535
B.123
C.12
1.2536
B.124
C.12
1.2537
B.125
C.12
1.2538
B.126
C.12
1.2539
B.127
C.12
1.2540
B.128
C.12
1.2541
B.129
C.12
1.2542
B.130
C.12
1.2543
B.131
C.12
1.2544
B.132
C.12
1.2545
B.133
C.12
1.2546
B.134
C.12
1.2547
B.135
C.12
1.2548
B.136
C.12
1.2549
B.137
C.12
1.2550
B.138
C.12
1.2551
B.139
C.12
1.2552
B.140
C.12
1.2553
B.141
C.12
1.2554
B.142
C.12
1.2555
B.143
C.12
1.2556
B.144
C.12
1.2557
B.145
C.12
1.2558
B.146
C.12
1.2559
B.147
C.12
1.2560
B.148
C.12
1.2561
B.149
C.12
1.2562
B.150
C.12
1.2563
B.151
C.12
1.2564
B.152
C.12
1.2565
B.153
C.12
1.2566
B.154
C.12
1.2567
B.155
C.12
1.2568
B.156
C.12
1.2569
B.157
C.12
1.2570
B.158
C.12
1.2571
B.159
C.12
1.2572
B.160
C.12
1.2573
B.161
C.12
1.2574
B.162
C.12
1.2575
B.163
C.12
1.2576
B.164
C.12
1.2577
B.165
C.12
1.2578
B.166
C.12
1.2579
B.167
C.12
1.2580
B.168
C.12
1.2581
B.169
C.12
1.2582
B.170
C.12
1.2583
B.171
C.12
1.2584
B.172
C.12
1.2585
B.173
C.12
1.2586
B.174
C.12
1.2587
B.175
C.12
1.2588
B.176
C.12
1.2589
B.177
C.12
1.2590
B.178
C.12
1.2591
B.179
C.12
1.2592
B.180
C.12
1.2593
B.181
C.12
1.2594
B.182
C.12
1.2595
B.183
C.12
1.2596
B.184
C.12
1.2597
B.185
C.12
1.2598
B.186
C.12
1.2599
B.187
C.12
1.2600
B.188
C.12
1.2601
B.189
C.12
1.2602
B.190
C.12
1.2603
B.191
C.12
1.2604
B.192
C.12
1.2605
B.193
C.12
1.2606
B.194
C.12
1.2607
B.195
C.12
1.2608
B.196
C.12
1.2609
B.197
C.12
1.2610
B.198
C.12
1.2611
B.199
C.12
1.2612
B.200
C.12
1.2613
B.201
C.12
1.2614
B.1
C.13
1.2615
B.2
C.13
1.2616
B.3
C.13
1.2617
B.4
C.13
1.2618
B.5
C.13
1.2619
B.6
C.13
1.2620
B.7
C.13
1.2621
B.8
C.13
1.2622
B.9
C.13
1.2623
B.10
C.13
1.2624
B.11
C.13
1.2625
B.12
C.13
1.2626
B.13
C.13
1.2627
B.14
C.13
1.2628
B.15
C.13
1.2629
B.16
C.13
1.2630
B.17
C.13
1.2631
B.18
C.13
1.2632
B.19
C.13
1.2633
B.20
C.13
1.2634
B.21
C.13
1.2635
B.22
C.13
1.2636
B.23
C.13
1.2637
B.24
C.13
1.2638
B.25
C.13
1.2639
B.26
C.13
1.2640
B.27
C.13
1.2641
B.28
C.13
1.2642
B.29
C.13
1.2643
B.30
C.13
1.2644
B.31
C.13
1.2645
B.32
C.13
1.2646
B.33
C.13
1.2647
B.34
C.13
1.2648
B.35
C.13
1.2649
B.36
C.13
1.2650
B.37
C.13
1.2651
B.38
C.13
1.2652
B.39
C.13
1.2653
B.40
C.13
1.2654
B.41
C.13
1.2655
B.42
C.13
1.2656
B.43
C.13
1.2657
B.44
C.13
1.2658
B.45
C.13
1.2659
B.46
C.13
1.2660
B.47
C.13
1.2661
B.48
C.13
1.2662
B.49
C.13
1.2663
B.50
C.13
1.2664
B.51
C.13
1.2665
B.52
C.13
1.2666
B.53
C.13
1.2667
B.54
C.13
1.2668
B.55
C.13
1.2669
B.56
C.13
1.2670
B.57
C.13
1.2671
B.58.
C.13
1.2672
B.59
C.13
1.2673
B.60
C.13
1.2674
B.61
C.13
1.2675
B.62
C.13
1.2676
B.63
C.13
1.2677
B.64
C.13
1.2678
B.65
C.13
1.2679
B.66
C.13
1.2680
B.67
C.13
1.2681
B.68
C.13
1.2682
B.69
C.13
1.2683
B.70
C.13
1.2684
B.71
C.13
1.2685
B.72
C.13
1.2686
B.73
C.13
1.2687
B.74
C.13
1.2688
B.75
C.13
1.2689
B.76
C.13
1.2690
B.77
C.13
1.2691
B.78
C.13
1.2692
B.79
C.13
1.2693
B.80
C.13
1.2694
B.81
C.13
1.2695
B.82
C.13
1.2696
B.83
C.13
1.2697
B.84
C.13
1.2698
B.85
C.13
1.2699
B.86
C.13
1.2700
B.87
C.13
1.2701
B.88
C.13
1.2702
B.89
C.13
1.2703
B.90
C.13
1.2704
B.91
C.13
1.2705
B.92
C.13
1.2706
B.93
C.13
1.2707
B.94
C.13
1.2708
B.95
C.13
1.2709
B.96
C.13
1.2710
B.97
C.13
1.2711
B.98
C.13
1.2712
B.99
C.13
1.2713
B.100
C.13
1.2714
B.101
C.13
1.2715
B.102
C.13
1.2716
B.103
C.13
1.2717
B.104
C.13
1.2718
B.105
C.13
1.2719
B.106
C.13
1.2720
B.107
C.13
1.2721
B.108
C.13
1.2722
B.109
C.13
1.2723
B.110
C.13
1.2724
B.111
C.13
1.2725
B.112
C.13
1.2726
B.113
C.13
1.2727
B.114
C.13
1.2728
B.115
C.13
1.2729
B.116
C.13
1.2730
B.117
C.13
1.2731
B.118
C.13
1.2732
B.119
C.13
1.2733
B.120
C.13
1.2734
B.121
C.13
1.2735
B.122
C.13
1.2736
B.123
C.13
1.2737
B.124
C.13
1.2738
B.125
C.13
1.2739
B.126
C.13
1.2740
B.127
C.13
1.2741
B.128
C.13
1.2742
B.129
C.13
1.2743
B.130
C.13
1.2744
B.131
C.13
1.2745
B.132
C.13
1.2746
B.133
C.13
1.2747
B.134
C.13
1.2748
B.135
C.13
1.2749
B.136
C.13
1.2750
B.137
C.13
1.2751
B.138
C.13
1.2752
B.139
C.13
1.2753
B.140
C.13
1.2754
B.141
C.13
1.2755
B.142
C.13
1.2756
B.143
C.13
1.2757
B.144
C.13
1.2758
B.145
C.13
1.2759
B.146
C.13
1.2760
B.147
C.13
1.2761
B.148
C.13
1.2762
B.149
C.13
1.2763
B.150
C.13
1.2764
B.151
C.13
1.2765
B.152
C.13
1.2766
B.153
C.13
1.2767
B.154
C.13
1.2768
B.155
C.13
1.2769
B.156
C.13
1.2770
B.157
C.13
1.2771
B.158
C.13
1.2772
B.159
C.13
1.2773
B.160
C.13
1.2774
B.161
C.13
1.2775
B.162
C.13
1.2776
B.163
C.13
1.2777
B.164
C.13
1.2778
B.165
C.13
1.2779
B.166
C.13
1.2780
B.167
C.13
1.2781
B.168
C.13
1.2782
B.169
C.13
1.2783
B.170
C.13
1.2784
B.171
C.13
1.2785
B.172
C.13
1.2786
B.173
C.13
1.2787
B.174
C.13
1.2788
B.175
C.13
1.2789
B.176
C.13
1.2790
B.177
C.13
1.2791
B.178
C.13
1.2792
B.179
C.13
1.2793
B.180
C.13
1.2794
B.181
C.13
1.2795
B.182
C.13
1.2796
B.183
C.13
1.2797
B.184
C.13
1.2798
B.185
C.13
1.2799
B.186
C.13
1.2800
B.187
C.13
1.2801
B.188
C.13
1.2802
B.189
C.13
1.2803
B.190
C.13
1.2804
B.191
C.13
1.2805
B.192
C.13
1.2806
B.193
C.13
1.2807
B.194
C.13
1.2808
B.195
C.13
1.2809
B.196
C.13
1.2810
B.197
C.13
1.2811
B.198
C.13
1.2812
B.199
C.13
1.2813
B.200
C.13
1.2814
B.201
C.13
1.2815
B.1
C.14
1.2816
B.2
C.14
1.2817
B.3
C.14
1.2818
B.4
C.14
1.2819
B.5
C.14
1.2820
B.6
C.14
1.2821
B.7
C.14
1.2822
B.8
C.14
1.2823
B.9
C.14
1.2824
B.10
C.14
1.2825
B.11
C.14
1.2826
B.12
C.14
1.2827
B.13
C.14
1.2828
B.14
C.14
1.2829
B.15
C.14
1.2830
B.16
C.14
1.2831
B.17
C.14
1.2832
B.18
C.14
1.2833
B.19
C.14
1.2834
B.20
C.14
1.2835
B.21
C.14
1.2836
B.22
C.14
1.2837
B.23
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1.2869
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1.2870
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C.14
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1.2874
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1.2875
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1.2876
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1.2910
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1.2915
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1.2917
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1.2920
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1.2921
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1.2922
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1.2923
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1.2925
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1.2926
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1.2927
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1.2928
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1.2930
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1.2931
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1.2932
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C.14
1.2933
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1.2934
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C.14
1.2935
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1.2936
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C.14
1.2937
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C.14
1.2938
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1.2939
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1.2940
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C.14
1.2941
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C.14
1.2942
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C.14
1.2943
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1.2944
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1.2945
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1.2949
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1.2955
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1.2956
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1.2960
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1.2967
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1.2989
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1.2990
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1.2991
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1.2992
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1.2993
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1.2995
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1.2996
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1.2997
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C.14
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C.14
1.3000
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1.3001
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1.3002
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1.3004
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C.14
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1.3007
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B.1
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1.3070
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1.3071
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C.15
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1.3075
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1.3076
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1.3077
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1.3078
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1.3081
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1.3082
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1.3084
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1.3086
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1.3087
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1.3088
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1.3090
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1.3091
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1.3092
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1.3093
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1.3095
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1.3097
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1.3100
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1.3101
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1.3102
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1.3103
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1.3104
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1.3105
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1.3106
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1.3107
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1.3108
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1.3110
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1.3111
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1.3112
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1.3116
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1.3117
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1.3119
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1.3120
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C.15
1.3123
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C.15
1.3124
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C.15
1.3125
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C.15
1.3126
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C.15
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C.15
1.3128
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C.15
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1.3133
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C.15
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C.15
1.3135
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C.15
1.3136
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C.15
1.3137
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C.15
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C.15
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C.15
1.3140
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C.15
1.3141
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1.3142
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C.15
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1.3144
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1.3190
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1.3192
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1.3196
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1.3200
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1.3201
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1.3202
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1.3204
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C.15
1.3205
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C.15
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C.15
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1.3208
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B.1
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C.16
1.3270
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C.16
1.3271
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1.3272
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1.3273
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C.16
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C.16
1.3275
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C.16
1.3276
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C.16
1.3277
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1.3278
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C.16
1.3279
B.63
C.16
1.3280
B.64
C.16
1.3281
B.65
C.16
1.3282
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C.16
1.3283
B.67
C.16
1.3284
B.68
C.16
1.3285
B.69
C.16
1.3286
B.70
C.16
1.3287
B.71
C.16
1.3288
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C.16
1.3289
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1.3290
B.74
C.16
1.3291
B.75
C.16
1.3292
B.76
C.16
1.3293
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C.16
1.3294
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C.16
1.3295
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C.16
1.3296
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1.3297
B.81
C.16
1.3298
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C.16
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1.3301
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C.16
1.3303
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C.16
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C.16
1.3305
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1.3306
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1.3318
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C.16
1.3333
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1.3334
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1.3335
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1.3336
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C.16
1.3339
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C.16
1.3340
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1.3341
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C.16
1.3342
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C.16
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C.16
1.3344
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C.16
1.3345
B.129
C.16
1.3346
B.130
C.16
1.3347
B.131
C.16
1.3348
B.132
C.16
1.3349
B.133
C.16
1.3350
B.134
C.16
1.3351
B.135
C.16
1.3352
B.136
C.16
1.3353
B.137
C.16
1.3354
B.138
C.16
1.3355
B.139
C.16
1.3356
B.140
C.16
1.3357
B.141
C.16
1.3358
B.142
C.16
1.3359
B.143
C.16
1.3360
B.144
C.16
1.3361
B.145
C.16
1.3362
B.146
C.16
1.3363
B.147
C.16
1.3364
B.148
C.16
1.3365
B.149
C.16
1.3366
B.150
C.16
1.3367
B.151
C.16
1.3368
B.152
C.16
1.3369
B.153
C.16
1.3370
B.154
C.16
1.3371
B.155
C.16
1.3372
B.156
C.16
1.3373
B.157
C.16
1.3374
B.158
C.16
1.3375
B.159
C.16
1.3376
B.160
C.16
1.3377
B.161
C.16
1.3378
B.162
C.16
1.3379
B.163
C.16
1.3380
B.164
C.16
1.3381
B.165
C.16
1.3382
B.166
C.16
1.3383
B.167
C.16
1.3384
B.168
C.16
1.3385
B.169
C.16
1.3386
B.170
C.16
1.3387
B.171
C.16
1.3388
B.172
C.16
1.3389
B.173
C.16
1.3390
B.174
C.16
1.3391
B.175
C.16
1.3392
B.176
C.16
1.3393
B.177
C.16
1.3394
B.178
C.16
1.3395
B.179
C.16
1.3396
B.180
C.16
1.3397
B.181
C.16
1.3398
B.182
C.16
1.3399
B.183
C.16
1.3400
B.184
C.16
1.3401
B.185
C.16
1.3402
B.186
C.16
1.3403
B.187
C.16
1.3404
B.188
C.16
1.3405
B.189
C.16
1.3406
B.190
C.16
1.3407
B.191
C.16
1.3408
B.192
C.16
1.3409
B.193
C.16
1.3410
B.194
C.16
1.3411
B.195
C.16
1.3412
B.196
C.16
1.3413
B.197
C.16
1.3414
B.198
C.16
1.3415
B.199
C.16
1.3416
B.200
C.16
1.3417
B.201
C.16
1.3418
B.1
C.17
1.3419
B.2
C.17
1.3420
B.3
C.17
1.3421
B.4
C.17
1.3422
B.5
C.17
1.3423
B.6
C.17
1.3424
B.7
C.17
1.3425
B.8
C.17
1.3426
B.9
C.17
1.3427
B.10
C.17
1.3428
B.11
C.17
1.3429
B.12
C.17
1.3430
B.13
C.17
1.3431
B.14
C.17
1.3432
B.15
C.17
1.3433
B.16
C.17
1.3434
B.17
C.17
1.3435
B.18
C.17
1.3436
B.19
C.17
1.3437
B.20
C.17
1.3438
B.21
C.17
1.3439
B.22
C.17
1.3440
B.23
C.17
1.3441
B.24
C.17
1.3442
B.25
C.17
1.3443
B.26
C.17
1.3444
B.27
C.17
1.3445
B.28
C.17
1.3446
B.29
C.17
1.3447
B.30
C.17
1.3448
B.31
C.17
1.3449
B.32
C.17
1.3450
B.33
C.17
1.3451
B.34
C.17
1.3452
B.35
C.17
1.3453
B.36
C.17
1.3454
B.37
C.17
1.3455
B.38
C.17
1.3456
B.39
C.17
1.3457
B.40
C.17
1.3458
B.41
C.17
1.3459
B.42
C.17
1.3460
B.43
C.17
1.3461
B.44
C.17
1.3462
B.45
C.17
1.3463
B.46
C.17
1.3464
B.47
C.17
1.3465
B.48
C.17
1.3466
B.49
C.17
1.3467
B.50
C.17
1.3468
B.51
C.17
1.3469
B.52
C.17
1.3470
B.53
C.17
1.3471
B.54
C.17
1.3472
B.55
C.17
1.3473
B.56
C.17
1.3474
B.57
C.17
1.3475
B.58.
C.17
1.3476
B.59
C.17
1.3477
B.60
C.17
1.3478
B.61
C.17
1.3479
B.62
C.17
1.3480
B.63
C.17
1.3481
B.64
C.17
1.3482
B.65
C.17
1.3483
B.66
C.17
1.3484
B.67
C.17
1.3485
B.68
C.17
1.3486
B.69
C.17
1.3487
B.70
C.17
1.3488
B.71
C.17
1.3489
B.72
C.17
1.3490
B.73
C.17
1.3491
B.74
C.17
1.3492
B.75
C.17
1.3493
B.76
C.17
1.3494
B.77
C.17
1.3495
B.78
C.17
1.3496
B.79
C.17
1.3497
B.80
C.17
1.3498
B.81
C.17
1.3499
B.82
C.17
1.3500
B.83
C.17
1.3501
B.84
C.17
1.3502
B.85
C.17
1.3503
B.86
C.17
1.3504
B.87
C.17
1.3505
B.88
C.17
1.3506
B.89
C.17
1.3507
B.90
C.17
1.3508
B.91
C.17
1.3509
B.92
C.17
1.3510
B.93
C.17
1.3511
B.94
C.17
1.3512
B.95
C.17
1.3513
B.96
C.17
1.3514
B.97
C.17
1.3515
B.98
C.17
1.3516
B.99
C.17
1.3517
B.100
C.17
1.3518
B.101
C.17
1.3519
B.102
C.17
1.3520
B.103
C.17
1.3521
B.104
C.17
1.3522
B.105
C.17
1.3523
B.106
C.17
1.3524
B.107
C.17
1.3525
B.108
C.17
1.3526
B.109
C.17
1.3527
B.110
C.17
1.3528
B.111
C.17
1.3529
B.112
C.17
1.3530
B.113
C.17
1.3531
B.114
C.17
1.3532
B.115
C.17
1.3533
B.116
C.17
1.3534
B.117
C.17
1.3535
B.118
C.17
1.3536
B.119
C.17
1.3537
B.120
C.17
1.3538
B.121
C.17
1.3539
B.122
C.17
1.3540
B.123
C.17
1.3541
B.124
C.17
1.3542
B.125
C.17
1.3543
B.126
C.17
1.3544
B.127
C.17
1.3545
B.128
C.17
1.3546
B.129
C.17
1.3547
B.130
C.17
1.3548
B.131
C.17
1.3549
B.132
C.17
1.3550
B.133
C.17
1.3551
B.134
C.17
1.3552
B.135
C.17
1.3553
B.136
C.17
1.3554
B.137
C.17
1.3555
B.138
C.17
1.3556
B.139
C.17
1.3557
B.140
C.17
1.3558
B.141
C.17
1.3559
B.142
C.17
1.3560
B.143
C.17
1.3561
B.144
C.17
1.3562
B.145
C.17
1.3563
B.146
C.17
1.3564
B.147
C.17
1.3565
B.148
C.17
1.3566
B.149
C.17
1.3567
B.150
C.17
1.3568
B.151
C.17
1.3569
B.152
C.17
1.3570
B.153
C.17
1.3571
B.154
C.17
1.3572
B.155
C.17
1.3573
B.156
C.17
1.3574
B.157
C.17
1.3575
B.158
C.17
1.3576
B.159
C.17
1.3577
B.160
C.17
1.3578
B.161
C.17
1.3579
B.162
C.17
1.3580
B.163
C.17
1.3581
B.164
C.17
1.3582
B.165
C.17
1.3583
B.166
C.17
1.3584
B.167
C.17
1.3585
B.168
C.17
1.3586
B.169
C.17
1.3587
B.170
C.17
1.3588
B.171
C.17
1.3589
B.172
C.17
1.3590
B.173
C.17
1.3591
B.174
C.17
1.3592
B.175
C.17
1.3593
B.176
C.17
1.3594
B.177
C.17
1.3595
B.178
C.17
1.3596
B.179
C.17
1.3597
B.180
C.17
1.3598
B.181
C.17
1.3599
B.182
C.17
1.3600
B.183
C.17
1.3601
B.184
C.17
1.3602
B.185
C.17
1.3603
B.186
C.17
1.3604
B.187
C.17
1.3605
B.188
C.17
1.3606
B.189
C.17
1.3607
B.190
C.17
1.3608
B.191
C.17
1.3609
B.192
C.17
1.3610
B.193
C.17
1.3611
B.194
C.17
1.3612
B.195
C.17
1.3613
B.196
C.17
1.3614
B.197
C.17
1.3615
B.198
C.17
1.3616
B.199
C.17
1.3617
B.200
C.17
1.3618
B.201
C.17
1.3619
—
C.1
1.3620
—
C.2
1.3621
—
C.3
1.3622
—
C.4
1.3623
—
C.5
1.3624
—
C.6
1.3625
—
C.7
1.3626
—
C.8
1.3627
—
C.9
1.3628
—
C.10
1.3629
—
C.11
1.3630
—
C.12
1.3631
—
C.13
1.3632
—
C.14
1.3633
—
C.15
1.3634
—
C.16
1.3635
—
C.17
1.3636
B.202
—
1.3637
B.202
C.1
1.3638
B.202
C.2
1.3639
B.202
C.3
1.3640
B.202
C.4
1.3641
B.202
C.5
1.3642
B.202
C.6
1.3643
B.202
C.7
1.3644
B.202
C.8
1.3645
B.202
C.9
1.3646
B.202
C.10
1.3647
B.202
C.11
1.3648
B.202
C.12
1.3649
B.202
C.13
1.3650
B.202
C.14
1.3651
B.202
C.15
1.3652
B.202
C.16
1.3653
B.202
C.17
The specific number for each single composition is deductible as follows: Composition 1.200 for example comprises the uracilpyridine I.a.339 and cinmethylin (B.200) (see table 1, entry 1.200; as well as table B, entry B.200).
Composition 2.200 for example comprises the uracilpyridine I.a.109 (see the definition for compositions 2.1 to 2.3653, preferably 2.1 to 2.3635, below) and cinmethylin (B.200) (see table 1, entry 1.200; as well as table B, entry B.200).
Composition 7.200 for example comprises imazapyr (B.35) (see the definition for compositions 7.1 to 7.3653, preferably 7.1 to 7.3635, below), the uracilpyridine I.a.339 and cinmethylin (B.200) (see table 1, entry 1.200; as well as table B, entry B.200).
Also especially preferred are compositions 2.1 to 2.3653, more preferred 2.1. to 2.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.109).
Also especially preferred are compositions 3.1 to 3.3653, more preferred 3.1. to 3.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.2 as further herbicide B.
Also especially preferred are compositions 4.1 to 4.3653, more preferred 4.1. to 4.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.8 as further herbicide B.
Also especially preferred are compositions 5.1 to 5.3653, more preferred 5.1. to 5.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.30 as further herbicide B.
Also especially preferred are compositions 6.1 to 6.3653, more preferred 6.1. to 6.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.32 as further herbicide B.
Also especially preferred are compositions 7.1 to 7.3653, more preferred 7.1. to 7.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.35 as further herbicide B.
Also especially preferred are compositions 8.1 to 8.3653, more preferred 8.1. to 8.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.38 as further herbicide B.
Also especially preferred are compositions 9.1 to 9.3653, more preferred 9.1. to 9.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.40 as further herbicide B.
Also especially preferred are compositions 10.1 to 10.3653, more preferred 10.1. to 10.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.51 as further herbicide B.
Also especially preferred are compositions 11.1 to 11.3653, more preferred 11.1. to 11.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.55 as further herbicide B.
Also especially preferred are compositions 12.1 to 12.3653, more preferred 12.1. to 12.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.56 as further herbicide B.
Also especially preferred are compositions 13.1 to 13.3653, more preferred 13.1. to 13.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.64 as further herbicide B.
Also especially preferred are compositions 14.1 to 14.3653, more preferred 14.1. to 14.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.66 as further herbicide B.
Also especially preferred are compositions 15.1 to 15.3653, more preferred 15.1. to 15.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.67 as further herbicide B.
Also especially preferred are compositions 16.1 to 16.3653, more preferred 16.1. to 16.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.68 as further herbicide B.
Also especially preferred are compositions 17.1 to 17.3653, more preferred 17.1. to 17.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.69 as further herbicide B.
Also especially preferred are compositions 18.1 to 18.3653, more preferred 18.1. to 18.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.73 as further herbicide B.
Also especially preferred are compositions 19.1 to 19.3653, more preferred 19.1. to 19.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.76 as further herbicide B.
Also especially preferred are compositions 20.1 to 20.3653, more preferred 20.1. to 20.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.81 as further herbicide B.
Also especially preferred are compositions 21.1 to 21.3653, more preferred 21.1. to 21.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.82 as further herbicide B.
Also especially preferred are compositions 22.1 to 22.3653, more preferred 22.1. to 22.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.85 as further herbicide B.
Also especially preferred are compositions 23.1 to 23.3653, more preferred 23.1. to 23.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.88 as further herbicide B.
Also especially preferred are compositions 24.1 to 24.3653, more preferred 24.1. to 24.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.89 as further herbicide B.
Also especially preferred are compositions 25.1 to 25.3653, more preferred 25.1. to 25.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.94 as further herbicide B.
Also especially preferred are compositions 26.1 to 26.3653, more preferred 26.1. to 26.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.95 as further herbicide B.
Also especially preferred are compositions 27.1 to 27.3653, more preferred 27.1. to 27.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.98 as further herbicide B.
Also especially preferred are compositions 28.1 to 28.3653, more preferred 28.1. to 28.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.100 as further herbicide B.
Also especially preferred are compositions 29.1 to 29.3653, more preferred 29.1. to 29.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.103 as further herbicide B.
Also especially preferred are compositions 30.1 to 30.3653, more preferred 30.1. to 30.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.103 and B.67 as further herbicides B.
Also especially preferred are compositions 31.1 to 31.3653, more preferred 31.1. to 31.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.103 and B.76 as further herbicides B.
Also especially preferred are compositions 32.1 to 32.3653, more preferred 32.1. to 32.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.103 and B.82 as further herbicides B.
Also especially preferred are compositions 33.1 to 33.3653, more preferred 33.1. to 33.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.104 as further herbicide B.
Also especially preferred are compositions 34.1 to 34.3653, more preferred 34.1. to 34.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.104 and B.67 as further herbicides B.
Also especially preferred are compositions 35.1 to 35.3653, more preferred 35.1. to 35.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.104 and B.76 as further herbicides B.
Also especially preferred are compositions 36.1 to 36.3653, more preferred 36.1. to 36.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.104 and B.82 as further herbicides B.
Also especially preferred are compositions 37.1 to 37.3653, more preferred 37.1. to 37.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.106 as further herbicide B.
Also especially preferred are compositions 38.1 to 38.3653, more preferred 38.1. to 38.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.107 as further herbicide B.
Also especially preferred are compositions 39.1 to 39.3653, more preferred 39.1. to 39.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B. 107 and B.67 as further herbicides B.
Also especially preferred are compositions 40.1 to 40.3653, more preferred 40.1. to 40.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B. 107 and B.76 as further herbicides B.
Also especially preferred are compositions 41.1 to 41.3653, more preferred 41.1. to 41.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B. 107 and B.82 as further herbicides B.
Also especially preferred are compositions 42.1 to 42.3653, more preferred 42.1. to 42.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.109 as further herbicide B.
Also especially preferred are compositions 43.1 to 43.3653, more preferred 43.1. to 43.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.111 as further herbicide B.
Also especially preferred are compositions 44.1 to 44.3653, more preferred 44.1. to 44.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.111 and B.67 as further herbicides B.
Also especially preferred are compositions 45.1 to 45.3653, more preferred 45.1. to 45.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.111 and B.76 as further herbicides B.
Also especially preferred are compositions 46.1 to 46.3653, more preferred 46.1. to 46.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.111 and B.82 as further herbicides B.
Also especially preferred are compositions 47.1 to 47.3653, more preferred 47.1. to 47.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B. 116 as further herbicide B.
Also especially preferred are compositions 48.1 to 48.3653, more preferred 48.1. to 48.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.67 as further herbicides B.
Also especially preferred are compositions 49.1 to 49.3653, more preferred 49.1. to 49.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.94 as further herbicides B.
Also especially preferred are compositions 50.1 to 50.3653, more preferred 50.1. to 50.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.103 as further herbicides B.
Also especially preferred are compositions 51.1 to 51.3653, more preferred 51.1. to 51.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.128 as further herbicides B.
Also especially preferred are compositions 52.1 to 52.3653, more preferred 52.1. to 52.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.104 as further herbicides B.
Also especially preferred are compositions 53.1 to 53.3653, more preferred 53.1. to 53.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.107 as further herbicides B.
Also especially preferred are compositions 54.1 to 54.3653, more preferred 54.1. to 54.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.116 and B.111 as further herbicides B.
Also especially preferred are compositions 55.1 to 55.3653, more preferred 55.1. to 55.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.122 as further herbicide B.
Also especially preferred are compositions 56.1 to 56.3653, more preferred 56.1. to 56.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.126 as further herbicide B.
Also especially preferred are compositions 57.1 to 57.3653, more preferred 57.1. to 57.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.128 as further herbicide B.
Also especially preferred are compositions 58.1 to 58.3653, more preferred 58.1. to 58.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.131 as further herbicide B.
Also especially preferred are compositions 59.1 to 59.3653, more preferred 59.1. to 59.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.132 as further herbicide B.
Also especially preferred are compositions 60.1 to 60.3653, more preferred 60.1. to 60.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.133 as further herbicide B.
Also especially preferred are compositions 61.1 to 61.3653, more preferred 61.1. to 61.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.135 as further herbicide B.
Also especially preferred are compositions 62.1 to 62.3653, more preferred 62.1. to 62.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.137 as further herbicide B.
Also especially preferred are compositions 63.1 to 63.3653, more preferred 63.1. to 63.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 11.1 to 1.3635, only in that they additionally comprise B.138 as further herbicide B.
Also especially preferred are compositions 64.1 to 64.3653, more preferred 64.1. to 64.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.140 as further herbicide B.
Also especially preferred are compositions 65.1 to 65.3653, more preferred 65.1. to 65.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.145 as further herbicide B.
Also especially preferred are compositions 66.1 to 66.3653, more preferred 66.1. to 66.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.153 as further herbicide B.
Also especially preferred are compositions 67.1 to 67.3653, more preferred 67.1. to 67.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.156 as further herbicide B.
Also especially preferred are compositions 68.1 to 68.3653, more preferred 68.1. to 68.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.171 as further herbicide B.
Also especially preferred are compositions 69.1 to 69.3653, more preferred 69.1. to 69.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they additionally comprise B.174 as further herbicide B.
Also especially preferred are compositions 70.1 to 70.3653, more preferred 70.1. to 70.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.115).
Also especially preferred are compositions 71.1 to 71.3653, more preferred 71.1. to 71.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.255).
Also especially preferred are compositions 72.1 to 72.3653, more preferred 72.1. to 72.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.277).
Also especially preferred are compositions 73.1 to 73.3653, more preferred 73.1. to 73.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.283).
Also especially preferred are compositions 74.1 to 74.3653, more preferred 74.1. to 74.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.87).
Also especially preferred are compositions 75.1 to 75.3653, more preferred 75.1. to 75.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.361).
Also especially preferred are compositions 76.1 to 76.3653, more preferred 76.1. to 76.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.a.367).
Also especially preferred are compositions 77.1 to 77.3653, more preferred 77.1. to 77.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.87).
Also especially preferred are compositions 78.1 to 78.3653, more preferred 78.1. to 78.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.109).
Also especially preferred are compositions 79.1 to 79.3653, more preferred 79.1. to 79.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.115).
Also especially preferred are compositions 80.1 to 80.3653, more preferred 80.1. to 80.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.255).
Also especially preferred are compositions 81.1 to 81.3653, more preferred 81.1. to 81.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.277).
Also especially preferred are compositions 82.1 to 82.3653, more preferred 82.1. to 82.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.283).
Also especially preferred are compositions 83.1 to 83.3653, more preferred 83.1. to 83.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339).
Also especially preferred are compositions 84.1 to 84.3653, more preferred 84.1. to 84.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.361).
Also especially preferred are compositions 85.1 to 85.3653, more preferred 85.1. to 85.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.367).
Also especially preferred are compositions 86.1 to 86.3653, more preferred 86.1. to 86.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.2 as further herbicide B.
Also especially preferred are compositions 87.1 to 87.3653, more preferred 87.1. to 87.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.8 as further herbicide B.
Also especially preferred are compositions 88.1 to 88.3653, more preferred 88.1. to 88.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.30 as further herbicide B.
Also especially preferred are compositions 89.1 to 89.3653, more preferred 89.1. to 89.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.32 as further herbicide B.
Also especially preferred are compositions 90.1 to 90.3653, more preferred 90.1. to 90.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.35 as further herbicide B.
Also especially preferred are compositions 91.1 to 91.3653, more preferred 91.1. to 91.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.38 as further herbicide B.
Also especially preferred are compositions 92.1 to 92.3653, more preferred 92.1. to 92.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.40 as further herbicide B.
Also especially preferred are compositions 93.1 to 93.3653, more preferred 93.1. to 93.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.51 as further herbicide B.
Also especially preferred are compositions 94.1 to 94.3653, more preferred 94.1. to 94.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.55 as further herbicide B.
Also especially preferred are compositions 95.1 to 95.3653, more preferred 95.1. to 95.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.56 as further herbicide B.
Also especially preferred are compositions 96.1 to 96.3653, more preferred 96.1. to 96.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.64 as further herbicide B.
Also especially preferred are compositions 97.1 to 97.3653, more preferred 97.1. to 97.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.66 as further herbicide B.
Also especially preferred are compositions 98.1 to 98.3653, more preferred 98.1. to 98.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.67 as further herbicide B.
Also especially preferred are compositions 99.1 to 99.3653, more preferred 99.1. to 99.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.68 as further herbicide B.
Also especially preferred are compositions 100.1 to 100.3653, more preferred 100.1. to 100.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.69 as further herbicide B.
Also especially preferred are compositions 101.1 to 101.3653, more preferred 101.1. to 101.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.73 as further herbicide B.
Also especially preferred are compositions 102.1 to 102.3653, more preferred 102.1. to 102.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635 only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.76 as further herbicide B.
Also especially preferred are compositions 103.1 to 103.3653, more preferred 103.1. to 103.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.81 as further herbicide B.
Also especially preferred are compositions 104.1 to 104.3653, more preferred 104.1. to 104.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.82 as further herbicide B.
Also especially preferred are compositions 105.1 to 105.3653, more preferred 105.1. to 105.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.85 as further herbicide B.
Also especially preferred are compositions 106.1 to 106.3653, more preferred 106.1. to 106.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.88 as further herbicide B.
Also especially preferred are compositions 107.1 to 107.3653, more preferred 107.1. to 107.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.89 as further herbicide B.
Also especially preferred are compositions 108.1 to 108.3653, more preferred 108.1. to 108.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.94 as further herbicide B.
Also especially preferred are compositions 191.1 to 109.3653, more preferred 109.1. to 109.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.95 as further herbicide B.
Also especially preferred are compositions 110.1 to 110.3653, more preferred 110.1. to 110.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.98 as further herbicide B.
Also especially preferred are compositions 111.1 to 111.3653, more preferred 111.1. to 111.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.100 as further herbicide B.
Also especially preferred are compositions 112.1 to 112.3653, more preferred 112.1. to 112.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.103 as further herbicide B.
Also especially preferred are compositions 113.1 to 113.3653, more preferred 113.1. to 113.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.103 and B.67 as further herbicides B.
Also especially preferred are compositions 114.1 to 114.3653, more preferred 114.1. to 114.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.103 and B.76 as further herbicides B.
Also especially preferred are compositions 115.1 to 115.3653, more preferred 115.1. to 115.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.103 and B.82 as further herbicides B.
Also especially preferred are compositions 116.1 to 116.3653, more preferred 116.1. to 116.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.104 as further herbicide B.
Also especially preferred are compositions 117.1 to 117.3653, more preferred 117.1. to 117.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.104 and B.67 as further herbicides B.
Also especially preferred are compositions 118.1 to 118.3653, more preferred 118.1. to 118.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.104 and B.76 as further herbicides B.
Also especially preferred are compositions 119.1 to 119.3653, more preferred 119.1. to 119.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.104 and B.82 as further herbicides B.
Also especially preferred are compositions 120.1 to 120.3653, more preferred 120.1. to 120.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.106 as further herbicide B.
Also especially preferred are compositions 121.1 to 121.3653, more preferred 121.1. to 121.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.107 as further herbicide B.
Also especially preferred are compositions 122.1 to 122.3653, more preferred 122.1. to 122.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B. 107 and B.67 as further herbicides B.
Also especially preferred are compositions 123.1 to 123.3653, more preferred 123.1. to 123.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B. 107 and B.76 as further herbicides B.
Also especially preferred are compositions 124.1 to 124.3653, more preferred 124.1. to 124.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B. 107 and B.82 as further herbicides B.
Also especially preferred are compositions 125.1 to 125.3653, more preferred 125.1. to 125.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.109 as further herbicide B.
Also especially preferred are compositions 126.1 to 126.3653, more preferred 126.1. to 126.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.111 as further herbicide B.
Also especially preferred are compositions 127.1 to 127.3653, more preferred 127.1. to 127.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.111 and B.67 as further herbicides B.
Also especially preferred are compositions 128.1 to 128.3653, more preferred 128.1. to 128.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.111 and B.76 as further herbicides B.
Also especially preferred are compositions 129.1 to 129.3653, more preferred 129.1. to 129.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.111 and B.82 as further herbicides B.
Also especially preferred are compositions 130.1 to 130.3653, more preferred 130.1. to 130.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B. 116 as further herbicide B.
Also especially preferred are compositions 131.1 to 131.3653, more preferred 131.1. to 131.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.67 as further herbicides B.
Also especially preferred are compositions 132.1 to 132.3653, more preferred 132.1. to 132.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.94 as further herbicides B.
Also especially preferred are compositions 133.1 to 133.3653, more preferred 133.1. to 133.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.103 as further herbicides B.
Also especially preferred are compositions 134.1 to 134.3653, more preferred 134.1. to 134.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.128 as further herbicides B.
Also especially preferred are compositions 135.1 to 135.3653, more preferred 135.1. to 135.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.104 as further herbicides B.
Also especially preferred are compositions 136.1 to 136.3653, more preferred 136.1. to 136.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.107 as further herbicides B.
Also especially preferred are compositions 137.1 to 137.3653, more preferred 137.1. to 137.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.116 and B.111 as further herbicides B.
Also especially preferred are compositions 138.1 to 138.3653, more preferred 138.1. to 138.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.122 as further herbicide B.
Also especially preferred are compositions 139.1 to 139.3653, more preferred 139.1. to 139.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.126 as further herbicide B.
Also especially preferred are compositions 140.1 to 140.3653, more preferred 140.1. to 140.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.128 as further herbicide B.
Also especially preferred are compositions 141.1 to 141.3653, more preferred 141.1. to 141.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.131 as further herbicide B.
Also especially preferred are compositions 142.1 to 142.3653, more preferred 142.1. to 142.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.132 as further herbicide B.
Also especially preferred are compositions 143.1 to 143.3653, more preferred 143.1. to 143.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.133 as further herbicide B.
Also especially preferred are compositions 144.1 to 144.3653, more preferred 144.1. to 144.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.135 as further herbicide B.
Also especially preferred are compositions 145.1 to 145.3653, more preferred 145.1. to 145.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.137 as further herbicide B.
Also especially preferred are compositions 146.1 to 146.3653, more preferred 146.1. to 146.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.138 as further herbicide B.
Also especially preferred are compositions 147.1 to 147.3653, more preferred 147.1. to 147.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.140 as further herbicide B.
Also especially preferred are compositions 148.1 to 148.3653, more preferred 148.1. to 148.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.145 as further herbicide B.
Also especially preferred are compositions 149.1 to 149.3653, more preferred 149.1. to 149.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.153 as further herbicide B.
Also especially preferred are compositions 150.1 to 150.3653, more preferred 150.1. to 150.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.156 as further herbicide B.
Also especially preferred are compositions 151.1 to 151.3653, more preferred 151.1. to 151.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.171 as further herbicide B.
Also especially preferred are compositions 152.1 to 152.3653, more preferred 152.1. to 152.3635, which differ from the corresponding compositions 1.1 to 1.3653, more preferred 1.1 to 1.3635, only in that they comprise as the active compound A the uracilpyridine of formula (I.h.339) and additionally comprise B.174 as further herbicide B.
It is generally preferred to use the compounds of the invention in combination with herbicides that are selective for the crop being treated and which complement the spectrum of weeds controlled by these compounds at the application rate employed. It is further generally preferred to apply the compounds of the invention and other complementary herbicides at the same time, either as a combination formulation or as a tank mix.
It is recognized that the polynucleotide molecules and polypeptides of the invention encompass polypeptides comprising an amino acid sequence that is sufficiently identical to the amino acid sequences set forth in SEQ ID Nos: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637. The term “sufficiently identical” is used herein to refer to a first amino acid or nucleotide sequence that contains a sufficient or minimum number of identical or equivalent (e.g., with a similar side chain) amino acid residues or nucleotides to a second amino acid or nucleotide sequence such that the first and second amino acid or nucleotide sequences have a common structural domain and/or common functional activity.
Generally, “sequence identity” refers to the extent to which two optimally aligned DNA or amino acid sequences are invariant throughout a window of alignment of components, e.g., nucleotides or amino acids. An “identity fraction” for aligned segments of a test sequence and a reference sequence is the number of identical components that are shared by the two aligned sequences divided by the total number of components in reference sequence segment, i.e., the entire reference sequence or a smaller defined part of the reference sequence. “Percent identity” is the identity fraction times 100. Optimal alignment of sequences for aligning a comparison window are well known to those skilled in the art and may be conducted by tools such as the local homology algorithm of Smith and Waterman, the homology alignment algorithm of Needleman and Wunsch, the search for similarity method of Pearson and Lipman, and preferably by computerized implementations of these algorithms such as GAP, BESTFIT, FASTA, and TFASTA available as part of the GCG. Wisconsin Package. (Accelrys Inc. Burlington, Mass.)
Polynucleotides and Oligonucleotides
By an “isolated polynucleotide”, including DNA, RNA, or a combination of these, single or double stranded, in the sense or antisense orientation or a combination of both, dsRNA or otherwise, we mean a polynucleotide which is at least partially separated from the polynucleotide sequences with which it is associated or linked in its native state. That means other nucleic acid molecules are present in an amount less than 5% based on weight of the amount of the desired nucleic acid, preferably less than 2% by weight, more preferably less than 1% by weight, most preferably less than 0.5% by weight. Preferably, an “isolated” nucleic acid is free of some of the sequences that naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated herbicide resistance and/or tolerance related protein encoding nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be free from some of the other cellular material with which it is naturally associated, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. Preferably, the isolated polynucleotide is at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally associated. As the skilled addressee would be aware, an isolated polynucleotide can be an exogenous polynucleotide present in, for example, a transgenic organism which does not naturally comprise the polynucleotide. Furthermore, the terms “polynucleotide(s)”, “nucleic acid sequence(s)”, “nucleotide sequence(s)”, “nucleic acid(s)”, “nucleic acid molecule” are used interchangeably herein and refer to nucleotides, either ribonucleotides or deoxyribonucleotides or a combination of both, in a polymeric unbranched form of any length.
The term “mutated PPO nucleic acid” refers to a PPO nucleic acid having a sequence that is mutated from a wild-type PPO nucleic acid, such as e.g. SEQ ID NO: 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 624, 626, or 650, or homologues, paralogues and orthologues thereof; and that confers increased uracilpyridine herbicide tolerance to a plant in which it is expressed. Furthermore, the term “mutated protoporphyrinogen oxidase (mutated PPO)” refers to the replacement of an amino acid of the wild-type primary sequences SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, or a variant, a derivative, a homologue, an orthologue, or paralogue thereof, with another amino acid. The expression “mutated amino acid” will be used below to designate the amino acid which is replaced by another amino acid, thereby designating the site of the mutation in the primary sequence of the protein.
In a preferred embodiment, the PPO nucleotide sequence encoding a mutated PPO comprises the sequence of SEQ ID NO: 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 624, 626, or 650, or a variant or derivative thereof.
Furthermore, it will be understood by the person skilled in the art that the PPO nucleotide sequences encompasse homologues, paralogues and orthologues of SEQ ID NO: 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 624, 626, or 650, as defined hereinafter.
The term “variant” with respect to a sequence (e.g., a polypeptide or nucleic acid sequence such as—for example—a transcription regulating nucleotide sequence of the invention) is intended to mean substantially similar sequences. For nucleotide sequences comprising an open reading frame, variants include those sequences that, because of the degeneracy of the genetic code, encode the identical amino acid sequence of the native protein. Naturally occurring allelic variants such as these can be identified with the use of well-known molecular biology techniques, as, for example, with polymerase chain reaction (PCR) and hybridization techniques. Variant nucleotide sequences also include synthetically derived nucleotide sequences, such as those generated, for example, by using site-directed mutagenesis and for open reading frames, encode the native protein, as well as those that encode a polypeptide having amino acid substitutions relative to the native protein, e.g. the mutated PPO according to the present invention as disclosed herein.
Generally, nucleotide sequence variants of the invention will have at least 30, 40, 50, 60, to 70%, e.g., preferably 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, to 79%, generally at least 80%, e.g., 81%-84%, at least 85%, e.g., 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, to 98% and 99% nucleotide “sequence identity” to the nucleotide sequence encoding a polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 637, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, or 565.
The % identity of a polynucleotide is determined by GAP (Needleman and Wunsch, 1970) analysis (GCG program) with a gap creation penalty=5, and a gap extension penalty=0.3. Unless stated otherwise, the query sequence is at least 45 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 45 nucleotides. Preferably, the query sequence is at least 150 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 150 nucleotides. More preferably, the query sequence is at least 300 nucleotides in length and the GAP analysis aligns the two sequences over a region of at least 300 nucleotides. Even more preferably, the GAP analysis aligns the two sequences over their entire length.
Polypeptides
By “substantially purified polypeptide” or “purified” a polypeptide is meant that has been separated from one or more lipids, nucleic acids, other polypeptides, or other contaminating molecules with which it is associated in its native state. It is preferred that the substantially purified polypeptide is at least 60% free, more preferably at least 75% free, and more preferably at least 90% free from other components with which it is naturally associated. As the skilled addressee will appreciate, the purified polypeptide can be a recombinantly produced polypeptide. The terms “polypeptide” and “protein” are generally used interchangeably and refer to a single polypeptide chain which may or may not be modified by addition of non-amino acid groups. It would be understood that such polypeptide chains may associate with other polypeptides or proteins or other molecules such as co-factors. The terms “proteins” and “polypeptides” as used herein also include variants, mutants, modifications, analogous and/or derivatives of the polypeptides of the invention as described herein.
The % identity of a polypeptide is determined by GAP (Needleman and Wunsch, 1970) analysis (GCG program) with a gap creation penalty=5, and a gap extension penalty=0.3. The query sequence is at least 25 amino acids in length, and the GAP analysis aligns the two sequences over a region of at least 25 amino acids. More preferably, the query sequence is at least 50 amino acids in length, and the GAP analysis aligns the two sequences over a region of at least 50 amino acids. More preferably, the query sequence is at least 100 amino acids in length and the GAP analysis aligns the two sequences over a region of at least 100 amino acids. Even more preferably, the query sequence is at least 250 amino acids in length and the GAP analysis aligns the two sequences over a region of at least 250 amino acids. Even more preferably, the GAP analysis aligns the two sequences over their entire length.
With regard to a defined polypeptide, it will be appreciated that % identity figures higher than those provided above will encompass preferred embodiments. Thus, where applicable, in light of the minimum % identity figures, it is preferred that the PPO polypeptide of the invention comprises an amino acid sequence which is at least 40%, more preferably at least 45%, more preferably at least 50%, more preferably at least 55%, more preferably at least 60%, more preferably at least 65%, more preferably at least 70%, more preferably at least 75%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 91%, more preferably at least 92%, more preferably at least 93%, more preferably at least 94%, more preferably at least 95%, more preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, more preferably at least 99.1%, more preferably at least 99.2%, more preferably at least 99.3%, more preferably at least 99.4%, more preferably at least 99.5%, more preferably at least 99.6%, more preferably at least 99.7%, more preferably at least 99.8%, and even more preferably at least 99.9% identical to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 637, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564,or 565.
By “variant” polypeptide is intended a polypeptide derived from the protein of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 637, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, or 565, by deletion (so-called truncation) or addition of one or more amino acids to the N-terminal and/or C-terminal end of the native protein; deletion or addition of one or more amino acids at one or more sites in the native protein; or substitution of one or more amino acids at one or more sites in the native protein. Such variants may result from, for example, genetic polymorphism or from human manipulation. Methods for such manipulations are generally known in the art.
“Derivatives” of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and/or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived.
“Homologues” of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and/or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived.
A deletion refers to removal of one or more amino acids from a protein.
An insertion refers to one or more amino acid residues being introduced into a predetermined site in a protein. Insertions may comprise N-terminal and/or C-terminal fusions as well as intra-sequence insertions of single or multiple amino acids. Generally, insertions within the amino acid sequence will be smaller than N- or C-terminal fusions, of the order of about 1 to 10 residues. Examples of N- or C-terminal fusion proteins or peptides include the binding domain or activation domain of a transcriptional activator as used in the yeast two-hybrid system, phage coat proteins, (histidine)-6-tag, glutathione S-transferase-tag, protein A, maltose-binding protein, dihydrofolate reductase, Tag•100 epitope, c-myc epitope, FLAG®-epitope, lacZ, CMP (calmodulin-binding peptide), HA epitope, protein C epitope and VSV epitope.
A substitution refers to replacement of amino acids of the protein with other amino acids having similar properties (such as similar hydrophobicity, hydrophilicity, antigenicity, propensity to form or break a-helical structures or β-sheet structures). Amino acid substitutions are typically of single residues, but may be clustered depending upon functional constraints placed upon the polypeptide and may range from 1 to 10 amino acids; insertions will usually be of the order of about 1 to 10 amino acid residues. The amino acid substitutions are preferably conservative amino acid substitutions. Conservative substitution tables are well known in the art (see for example Creighton (1984) Proteins. W.H. Freeman and Company (Eds).
TABLE 1
Examples of conserved amino acid substitutions
Conservative
Residue
Substitutions
Ala
Ser
Arg
Lys
Asn
Gln; His
Asp
Glu
Gln
Asn
Cys
Ser
Glu
Asp
Gly
Pro
His
Asn; Gln
Ile
Leu, Val
Leu
Ile; Val
Lys
Arg; Gln
Met
Leu; Ile
Phe
Met; Leu; Tyr
Ser
Thr; Gly
Thr
Ser; Val
Trp
Tyr
Tyr
Trp; Phe
Val
Ile; Leu
Amino acid substitutions, deletions and/or insertions may readily be made using peptide synthetic techniques well known in the art, such as solid phase peptide synthesis and the like, or by recombinant DNA manipulation. Methods for the manipulation of DNA sequences to produce substitution, insertion or deletion variants of a protein are well known in the art. For example, techniques for making substitution mutations at predetermined sites in DNA are well known to those skilled in the art and include M13 mutagenesis, T7-Gen in vitro mutagenesis (USB, Cleveland, Ohio), QuickChange Site Directed mutagenesis (Stratagene, San Diego, Calif.), PCR-mediated site-directed mutagenesis or other site-directed mutagenesis protocols.
“Derivatives” further include peptides, oligopeptides, polypeptides which may, compared to the amino acid sequence of the naturally-occurring form of the protein, such as the protein of interest, comprise substitutions of amino acids with non-naturally occurring amino acid residues, or additions of non-naturally occurring amino acid residues. “Derivatives” of a protein also encompass peptides, oligopeptides, polypeptides which comprise naturally occurring altered (glycosylated, acylated, prenylated, phosphorylated, myristoylated, sulphated etc.) or non-naturally altered amino acid residues compared to the amino acid sequence of a naturally-occurring form of the polypeptide. A derivative may also comprise one or more non-amino acid substituents or additions compared to the amino acid sequence from which it is derived, for example a reporter molecule or other ligand, covalently or non-covalently bound to the amino acid sequence, such as a reporter molecule which is bound to facilitate its detection, and non-naturally occurring amino acid residues relative to the amino acid sequence of a naturally-occurring protein. Furthermore, “derivatives” also include fusions of the naturally-occurring form of the protein with tagging peptides such as FLAG, HIS6 or thioredoxin (for a review of tagging peptides, see Terpe, Appl. Microbiol. Biotechnol. 60, 523-533, 2003).
“Orthologues” and “paralogues” encompass evolutionary concepts used to describe the ancestral relationships of genes. Paralogues are genes within the same species that have originated through duplication of an ancestral gene; orthologues are genes from different organisms that have originated through speciation, and are also derived from a common ancestral gene.
It is well-known in the art that paralogues and orthologues may share distinct domains harboring suitable amino acid residues at given sites, such as binding pockets for particular substrates or binding motifs for interaction with other proteins.
The term “domain” refers to a set of amino acids conserved at specific positions along an alignment of sequences of evolutionarily related proteins. While amino acids at other positions can vary between homologues, amino acids that are highly conserved at specific positions indicate amino acids that are likely essential in the structure, stability or function of a protein. Identified by their high degree of conservation in aligned sequences of a family of protein homologues, they can be used as identifiers to determine if any polypeptide in question belongs to a previously identified polypeptide family.
The term “motif” or “consensus sequence” refers to a short conserved region in the sequence of evolutionarily related proteins. Motifs are frequently highly conserved parts of domains, but may also include only part of the domain, or be located outside of conserved domain (if all of the amino acids of the motif fall outside of a defined domain). Preferred motifs which are contained in the sequences of the PPO polypeptides according to the present invention are SQ[N/K/H]KRYI, TLGTLFSS, [F/Y]TTF[V/I]GG.
Specialist databases exist for the identification of domains, for example, SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic et al. (2002) Nucleic Acids Res 30, 242-244), InterPro (Mulder et al., (2003) Nucl. Acids. Res. 31, 315-318), Prosite (Bucher and Bairoch (1994), A generalized profile syntax for biomolecular sequences motifs and its function in automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology. Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., pp 53-61, AAAI Press, Menlo Park; Hulo et al., Nucl. Acids. Res. 32:D134-D137, (2004)), or Pfam (Bateman et al., Nucleic Acids Research 30(1): 276-280 (2002)). A set of tools for in silico analysis of protein sequences is available on the ExPASy proteomics server (Swiss Institute of Bioinformatics (Gasteiger et al., ExPASy: the proteomics server for in-depth protein knowledge and analysis, Nucleic Acids Res. 31:3784-3788(2003)). Domains or motifs may also be identified using routine techniques, such as by sequence alignment.
Methods for the alignment of sequences for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the global (i.e. spanning the complete sequences) alignment of two sequences that maximizes the number of matches and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The software for performing BLAST analysis is publicly available through the National Centre for Biotechnology Information (NCBI).
Homologues may readily be identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the default pairwise alignment parameters, and a scoring method in percentage. Global percentages of similarity and identity may also be determined using one of the methods available in the MatGAT software package (Campanella et al., BMC Bioinformatics. 2003 Jul. 10; 4:29. MatGAT: an application that generates similarity/identity matrices using protein or DNA sequences.). Minor manual editing may be performed to optimise alignment between conserved motifs, as would be apparent to a person skilled in the art. Furthermore, instead of using full-length sequences for the identification of homologues, specific domains may also be used. The sequence identity values may be determined over the entire nucleic acid or amino acid sequence or over selected domains or conserved motif(s), using the programs mentioned above using the default parameters. For local alignments, the Smith-Waterman algorithm is particularly useful (Smith T F, Waterman M S (1981) J. Mol. Biol 147(1); 195-7).
The inventors of the present invention have found that by substituting one or more of the key amino acid residues, employing e.g. one of the above described methods to mutate the encoding nucleic acids, the uracilpyridine herbicide tolerance or resistance could be remarkably increased as compared to the activity of the wild type PPO enzymes with SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637. Preferred substitutions of mutated PPO are those that increase the herbicide tolerance of the plant, but leave the biological activity of the oxidase activity substantially unaffected.
Accordingly, in another object of the present invention the key amino acid residues of a PPO enzyme comprising SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637,a variant, derivative, orthologue, paralogue or homologue thereof, is substituted by any other amino acid.
In one embodiment, the key amino acid residues of a PPO enzyme, a variant, derivative, orthologue, paralogue or homologue thereof, is substituted by a conserved amino acid as depicted in Table 1.
It will be understood by the person skilled in the art that amino acids located in a close proximity to the positions of amino acids mentioned below may also be substituted. Thus, in another embodiment the variant of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof comprises a mutated PPO, wherein an amino acid 3, ±2 or ±1 amino acid positions from a key amino acid is substituted by any other amino acid.
Based on techniques well-known in the art, a highly characteristic sequence pattern can be developed, by means of which further of mutated PPO candidates with the desired activity may be searched.
Searching for further mutated PPO candidates by applying a suitable sequence pattern would also be encompassed by the present invention. It will be understood by a skilled reader that the present sequence pattern is not limited by the exact distances between two adjacent amino acid residues of said pattern. Each of the distances between two neighbours in the above patterns may, for example, vary independently of each other by up to ±10, ±5, ±3, ±2 or ±1 amino acid positions without substantially affecting the desired activity.
Furthermore, by applying the method of site directed mutagenesis, in particular saturation mutagenes (see e.g. Schenk et al., Biospektrum 03/2006, pages 277-279), the inventors of the present invention have identified and generated specific amino acid substitutions and combinations thereof, which—when introduced into a plant by transforming and expressing the respective mutated PPO encoding nucleic acid—confer increased herbicide resistance or tolerance to a uracilpyridine herbicide to said plant.
Thus, in a particularly preferred embodiment, the variant or derivative of the mutated PPO refers to a polypeptide comprising SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, or 637, comprising a single amino acid substitution at positions depicted in the following Table 2a.
TABLE 2a-1
Single amino acid substitutions within SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,
35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,
59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104,
105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, or 637
SEQ
Mutated
Mutated
Mutated
Mutated
Mutated
ID
Organism
site 1
site 2
site 3
site 4
site 5
1
Amaranthus tuberculatus
R128
G210
G211
L397
F420
2
Amaranthus tuberculatus
R128
G210
G211
L397
F420
3
Amaranthus tuberculatus
R128
G210
—
L396
F419
4
Amaranthus tuberculatus
R128
G210
—
L396
F419
5
Amaranthus
R128
G210
G211
L398
F421
hypochondriacus
6
Amaranthus tuberculatus
R98
G180
G181
L368
F391
7
Spinacia oleracea
R127
G209
G210
L397
F420
8
Vitis vinifera
R100
G182
G183
L373
F396
9
Ricinus communis
R99
A181
G182
L372
F395
10
Theobroma cacao
R98
A180
G181
L370
F393
11
Glycine max
R98
A180
A181
L367
F390
12
Prunus persica
R96
A175
G176
L366
F389
13
Medicago truncatula
R98
A180
A181
L367
F390
14
Fragaria vesca subsp. vesca
R98
A177
G178
L368
F391
15
Citrus clementina
R98
A180
A181
L370
F393
16
Citrus clementina
R98
A180
A181
L370
F393
17
Cicer arietinum
R98
A180
A181
L367
F390
18
Cucumis sativus
R98
A180
G181
L368
F391
19
Cucumis sativus
R93
A175
G176
L363
F386
20
Nicotiana tabacum
R98
G178
G179
L369
F392
21
Solanum lycopersicum
R95
G175
G176
L366
F389
22
Arabidopsis thaliana
R101
A182
A183
L371
F394
23
Arabidopsis lyrata subsp.
R103
A186
A187
L375
F398
lyrata
24
Arabidopsis thaliana
R101
A182
A183
L364
F387
25
Arabidopsis thaliana
R147
A228
A229
L410
F433
26
Ambrosia artemisiifolia
R74
G151
G152
L336
F359
27
Setaria italica
R128
A213
G214
L408
F431
28
Sorghum bicolor
R130
A215
G216
L410
F433
29
Arabidopsis thaliana
R101
A152
A153
L341
F364
30
Zea mays
R130
A215
G216
L410
F433
31
Zea mays
R130
A215
G216
L410
F433
32
LEMPA
R105
G188
G189
L381
F404
33
LEMPA
R150
A226
G227
L409
Y432
34
Populus trichocarpa
R100
A182
G183
L373
F396
35
Capsella rubella
R165
A248
A249
L433
F456
36
Brachypodium
R134
A219
G220
L414
F437
distachyon
37
Oryza sativa Japonica
R95
G179
G180
L374
F397
Group
38
Picea sitchensis
R100
G186
A187
L354
F377
39
Solanum tuberosum
R98
G178
G179
L369
F392
40
Oryza sativa Indica
R139
G224
G225
L419
F442
Group
41
Oryza sativa Japonica
R139
G224
G225
L419
F442
Group
42
Eutrema salsugineum
R96
A179
A180
L368
F391
43
Selaginella moellendorffii
R97
G175
S176
L377
F400
44
Selaginella moellendorffii
R97
G175
S176
L377
F400
45
Amaranthus tuberculatus
R128
G210
G211
—
—
46
Amaranthus tuberculatus
R128
G210
—
—
—
47
Zea mays
—
A67
G68
L262
F285
48
Aegilops tauschii
R30
A96
G97
L291
F314
49
Genlisea aurea
R89
G168
G169
—
—
50
Amborella trichopoda
—
—
—
L161
F184
51
Rhodothermus marinus
R87
A162
G163
L330
F353
52
Salinibacter ruber
R87
A161
G162
L330
F353
53
Salinibacter ruber M8
R87
A161
G162
L330
F353
54
Zea mays
—
—
—
L69
F92
55
Rhodothermus marinus
R87
A162
G163
L330
F353
56
Caldithrix abyssi
R86
A160
G161
L329
F352
57
Opitutus terrae PB90-1
R103
A178
G179
L345
M368
58
Verrucomicrobia
R91
A166
G167
L332
Y355
bacterium
59
Ignavibacterium album
R88
A163
G164
L332
F355
60
Coraliomargarita sp.
R88
G163
A164
L331
Y354
CAG:312
61
Salisaeta longa
R88
A163
G164
L329
F352
62
Ambrosia artemisiifolia
R98
G175
G176
—
—
63
Melioribacter roseus P3M-2
R88
A163
G164
L333
F356
64
Halothiobacillus
R97
A170
G171
L343
F366
neapolitanus c2
65
Chondrus crispus
Y98
S176
G177
L352
Y376
66
Rubritalea marina
R88
A163
G164
L339
M362
67
Acidobacteria bacterium
R88
A161
G162
L330
F353
68
Coraliomargarita
R88
A163
G164
L323
F346
akajimensis DSM 45221
69
Oscillochloris trichoides
R90
S164
G165
L340
L363
DG6
70
Opitutaceae bacterium
R86
A161
G162
L352
L375
TAV1
71
Amborella trichopoda
R87
A173
A174
L230
—
72
Opitutaceae bacterium
R105
A180
G181
L368
L391
TAV5
73
Chloroflexus sp. Y-400-fl
R91
A166
G167
L335
L358
74
Leptospirillum sp. Group II
R92
A167
S168
L335
F358
‘5-way CG’
75
Leptospirillum ferriphilum
R92
A167
S168
L335
F358
ML-04
76
Verrucomicrobia
R89
A165
A166
L334
Y357
bacterium SCGC
AAA300-O17
77
Chloroflexus aggregans
R92
A167
G168
L336
L359
DSM 9485
78
Desulfurobacterium
R86
A161
G162
L333
M356
thermolithotrophum
79
Desulfurobacterium sp.
R90
A165
G166
L332
M355
TCS-1
80
Arthrospira platensis C1
R93
A176
G177
L352
Y375
81
Leptospirillum sp. Group II
R92
A167
S168
L335
F358
‘C75’
82
Verrucomicrobiae
R87
A164
G165
L332
M355
bacterium DG1235
83
Verrucomicrobia
R89
A165
A166
L334
F357
bacterium SCGC
AAA300-K03
84
Synechococcus sp. JA-3-3Ab
R101
A177
G178
L350
F373
85
Hymenobacter norwichensis
R86
A161
G162
A321
F344
86
Pontibacter sp. BAB1700
R85
G159
G160
S319
Y342
87
Leptospirillum
R92
A172
S173
L339
F362
ferrodiazotrophum
88
Prevotella histicola
C89
A164
G165
L328
F351
F0411
89
Flexithrix dorotheae
R84
A158
G159
A313
F336
90
Geobacter
R93
A168
G169
L342
M365
metallireducens GS-15
91
Synechococcus sp. JA-2-
R93
A169
G170
L362
F385
3B′a(2-13)
92
Crinalium epipsammum
R85
A168
G169
L344
F367
PCC 9333
93
Planctomyces mans
A99
T187
S188
F358
F381
94
Geobacter
R93
A170
G171
L344
M367
uraniireducens Rf4
95
Acidithiobacillus
R87
A162
G163
L323
F346
ferrivorans
96
Prevotella melaninogenica
C88
A163
G164
L327
F350
97
Thermovibrio ammonificans
R86
A160
G161
L333
M356
98
Brassica_rapa
R143
A219
G220
L402
Y425
99
Brassica_rapa
R112
A195
A196
L384
F407
100
Gossypium
R146
A222
G223
L405
Y428
101
Gossypium
R98
A180
G181
L370
F393
102
Conyza_canadensis
R142
A218
G219
L401
Y424
103
Conyza_canadensis
R102
G179
G180
L365
F388
104
Kochia_scobaria
R172
A248
G249
L431
F454
105
Lolium_rigidum
R138
A214
G215
L397
Y420
106
Lolium_rigidum
R97
A182
G183
L377
F400
107
Gossypium hirsutum PPO1
R146
A222
G223
L405
Y428
108
Beta vulgaris PPO1
R167
A243
G244
L426
Y449
109
Hordeum vulgare PPO1
R137
A213
G214
L396
Y419
110
Hordeum vulgare PPO2
R142
A227
G228
L422
F445
111
Triticum aestivum PPO1
R138
A214
G215
L397
Y420
112
Solanum lycopersicum PPO2
R95
G175
G176
L366
F389
113
Triticum aestivum PPO1_v2
R153
A229
G230
L412
Y435
114
Gossypium hirsutum PPO1_v2
R146
A222
G223
L405
Y428
115
Gossypium hirsutum PPO2
R98
A180
G181
L370
F393
116
Beta vulgaris PPO1_v2
R167
A243
G244
L426
Y449
117
Brassica napus_PPO2
R99
A182
A183
L371
F394
637
Oryza sativa_PPO2
R139
G225
L419
F442
TABLE 2a-2
Additional single amino acid substitutions within
reference to SEQ ID NO: 1, or 637
SEQ
Mutated
Mutated
ID
Organism
site 6
site 7
1
Amaranthus tuberculatus
G398
L400
637
Oryza sativa_PPO2
G420
L422
In a further particularly preferred embodiment, the variant or derivative of the mutated PPO refers to SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, or 637, comprising a single amino acid substitutions at the positions depicted in the following Table 2b.
It should be noted that Mutated site 1 of Table 2a) corresponds to Pos 1 of Table 2b); Mutated site 2 of Table 2a) corresponds to Pos 16 of Table 2b); Mutated site 3 of Table 2a) corresponds to Pos 17 of Table 2b); Mutated site 4 of Table 2a) corresponds to Pos 38 of Table 2b); Mutated site 5 of Table 2a) corresponds to Pos 42 of Table 2b).
TABLE 2b
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
1
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
G211
L216
M218
H219
2
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
G211
L216
M218
H219
3
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
—
L215
M217
H218
4
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
—
L215
M217
Y218
5
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
G211
L216
V218
H219
6
R98
Y99
A101
S119
I121
A124
P134
K139
E152
S153
E159
F166
D172
C179
G180
G181
L186
M188
H189
7
R127
Y128
A130
S148
I150
A153
P163
K168
E181
S182
E188
F195
D201
S208
G209
G210
L215
M217
R218
8
R100
Y101
V103
S121
I123
A126
P136
K141
E154
S155
Q161
V168
D174
S181
G182
G183
L188
M190
H191
9
R99
Y100
V102
S120
I122
A125
P135
K140
E153
S154
Q160
V167
D173
S180
A181
G182
L187
V189
C190
10
R98
Y99
V101
S119
I121
A124
P134
K139
E152
S153
Q159
V166
D172
S179
A180
G181
L186
M188
R189
11
R98
Y99
V101
S119
L121
A124
P134
R139
E152
S153
E159
V166
D172
S179
A180
A181
L186
M188
R189
12
R96
Y97
V99
S117
F119
A122
P132
D137
E147
S148
Q154
V161
D167
S174
A175
G176
L181
M183
R184
13
R98
Y99
V101
S119
L121
A124
P134
R139
E152
S153
E159
V166
D172
S179
A180
A181
L186
I188
R189
14
R98
Y99
A101
S119
F121
T124
P134
K139
E149
S150
Q156
V163
D169
S176
A177
G178
L183
M185
P186
15
R98
Y99
V101
S119
F121
A124
P134
K139
E152
S153
Q159
V166
D172
S179
A180
A181
L186
M188
R189
16
R98
Y99
V101
S119
F121
A124
P134
K139
E152
S153
Q159
V166
D172
S179
A180
A181
L186
M188
R189
17
R98
Y99
V101
S119
L121
A124
P134
R139
E152
S153
E159
V166
D172
S179
A180
A181
L186
M188
R189
18
R98
Y99
V101
S119
F121
A124
P134
K139
E152
S153
Q159
V166
D172
S179
A180
G181
L186
M188
S189
19
R93
Y94
V96
S114
F116
A119
P129
K134
E147
S148
Q154
V161
D167
S174
A175
G176
L181
M183
S184
20
R98
Y99
A101
S119
F121
T124
P134
N139
E150
S151
Q157
V164
D170
C177
G178
G179
L184
M186
H187
21
R95
Y96
A98
S116
F118
T121
P131
N136
E147
S148
Q154
V161
D167
C174
G175
G176
L181
M183
H184
22
R101
Y102
V104
S122
V124
T127
P137
K142
E154
S155
Q161
V168
D174
S181
A182
A183
L188
M190
K191
23
R103
Y104
V106
S124
V126
T129
P139
K144
E158
S159
Q165
V172
D178
S185
A186
A187
L192
M194
K195
24
R101
Y102
V104
S122
V124
T127
P137
K142
E154
S155
Q161
V168
D174
S181
A182
A183
L188
M190
K191
25
R147
Y148
V150
S168
V170
T173
P183
K188
E200
S201
Q207
V214
D220
S227
A228
A229
L234
M236
K237
26
R74
Y75
V77
S95
F97
T100
P110
K115
E123
S124
Q130
V137
D143
S150
G151
G152
L157
M159
R160
27
R128
Y129
V131
S149
V151
T154
P164
K169
E185
S186
E192
V199
D205
S212
A213
G214
L219
I221
R222
28
R130
Y131
V133
S151
V153
T156
P166
K171
E187
S188
E194
V201
D207
S214
A215
G216
L221
I223
C224
29
R101
Y102
V104
—
—
—
—
—
E124
S125
Q131
V138
D144
S151
A152
A153
L158
M160
K161
30
R130
Y131
V133
S151
V153
T156
P166
K171
E187
S188
E194
V201
D207
S214
A215
G216
L221
I223
R224
31
R130
Y131
V133
S151
V153
T156
P166
K171
E187
S188
E194
V201
D207
S214
A215
G216
L221
I223
R224
32
R105
Y106
V108
S126
L128
T131
P141
R146
E160
S161
Q167
V174
D180
S187
G188
G189
L194
M196
P197
33
R150
F151
L153
F171
L173
I176
A186
P191
E201
E202
—
V212
E218
Y225
A226
G227
L232
M234
K235
34
R100
Y101
V103
S121
F123
A126
P136
K141
E154
S155
Q161
V168
D174
S181
A182
G183
L188
A190
R191
35
R165
Y166
V168
S186
V188
T191
P201
K206
E220
S221
R227
V234
D240
S247
A248
A249
L254
M277
K278
36
R134
Y135
V137
S155
V157
T160
P170
K175
E191
S192
E198
V205
D211
S218
A219
G220
L225
I227
R228
37
R95
Y96
V98
S116
V118
T121
P131
K136
E152
S153
C158
V165
D171
S178
G179
G180
L185
I187
R188
38
R100
Y101
V103
S121
T123
A126
P136
H141
E158
S159
R165
V172
D178
A185
G186
A187
L192
I194
R195
39
R98
Y99
A101
S119
F121
T124
P134
N139
E150
S151
Q157
V164
D170
C177
G178
G179
L184
M186
H187
40
R139
Y140
V142
S160
V162
T165
P175
K180
E196
S197
C213
V220
D229
S233
G234
G235
L240
I242
R243
41
R139
Y140
V142
S160
V162
T165
P175
K180
E196
S197
C213
V220
D229
S233
G234
G235
L240
I242
R243
42
R96
Y97
V99
S117
V119
T122
P132
K137
E151
S152
Q158
V165
D171
S178
A179
A180
L185
M187
K188
43
R97
Y98
V100
T118
L120
A123
P133
R138
E147
S148
E154
I161
D167
S174
G175
S176
I181
I183
R184
44
R97
Y98
V100
T118
L120
A123
P133
R138
E147
S148
E154
I161
D167
S174
G175
S176
I181
I183
R184
45
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
G211
L216
M218
H219
46
R128
Y129
A131
S149
I151
A154
P164
K169
E182
S183
E189
F196
D202
C209
G210
—
L215
M217
H218
47
—
—
—
S3
V5
T8
P18
K23
E39
S40
E46
V53
D59
S66
A67
G68
L73
I75
R76
48
R30
Y31
V33
—
—
—
P47
K52
E68
S69
E75
V82
D88
S95
A96
G97
L102
I104
R105
49
R89
Y90
V92
S110
I112
S115
P125
Q130
E140
S141
Q147
V154
D160
S167
G168
G169
L174
M176
R177
50
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
51
R87
F88
V90
T108
L110
P113
P123
H126
A137
K138
—
V148
E154
F161
A162
G163
L168
V170
R171
52
R87
Y88
V90
T108
L110
T113
P123
G126
A136
R137
—
V147
A153
F160
A161
G162
L167
V169
Q170
53
R87
Y88
V90
T108
L110
T113
P123
G126
A136
R137
—
V147
A153
F160
A161
G162
L167
V169
Q170
54
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
55
R87
F88
V90
T108
L110
P113
P123
H126
A137
K138
—
V148
E154
F161
A162
G163
L168
V170
R171
56
R86
Y87
V89
T107
L109
W112
P122
P125
A135
D136
—
F146
N152
Y159
A160
G161
L166
A168
P169
57
R103
Y104
V106
S124
L126
P129
L139
R142
A153
E154
—
F164
N170
Y177
A178
G179
L184
A186
R187
58
R91
Y92
I94
T112
L114
L117
P127
A130
A141
A142
—
F152
N158
Y165
A166
G167
L172
V174
Q175
59
R88
Y89
L91
T109
L111
A114
P124
G127
A138
E139
—
F149
N155
Y162
A163
G164
L169
V171
K172
60
R88
F89
A91
T109
L111
F114
P124
K127
A138
D139
—
V149
N155
Y162
G163
A164
L169
I171
K172
61
R88
F89
V91
T109
L111
T114
P124
A127
A138
S139
—
V149
D155
F162
A163
G164
L169
L171
K172
62
R98
Y99
V101
S119
F121
T124
P134
K139
E147
S148
Q154
V161
D167
S174
G175
G176
—
—
—
63
R88
Y89
L91
T109
L111
S114
P124
S127
A138
E139
—
F149
D155
F162
A163
G164
L169
V171
K172
64
R97
F98
A100
S118
L120
M123
A133
K136
A145
E146
—
F156
D162
Y169
A170
G171
L176
V178
Q179
65
Y98
Y99
M101
T119
L121
W124
P134
L139
S149
V150
Q155
V162
D168
Y175
S176
G177
L182
M184
K185
66
R88
F89
I91
S109
I111
L114
P124
K127
A138
D139
—
P149
N155
Y162
A163
G164
L169
V171
E172
67
R88
Y89
V91
S109
L111
W114
P124
G127
A136
D137
—
F147
N153
Y160
A161
G162
L167
V169
R170
68
R88
Y89
V91
T109
L111
I114
P124
K127
A138
D139
—
L149
N155
Y162
A163
G164
L169
L171
R172
69
R90
Y91
L93
M111
H113
M116
P126
P129
A139
S140
—
V150
D156
Y163
S164
G165
M170
I172
K173
70
R86
F87
V89
T107
L109
L112
L122
R125
G136
E137
—
L147
D153
Y160
A161
G162
L167
A169
R170
71
R87
F88
A90
S108
L110
P113
P123
S128
E145
S146
Q152
V159
D165
S172
A173
A174
L179
—
—
72
R105
F106
V108
T126
L128
L131
L141
R144
G155
E156
—
L166
D172
Y179
A180
G181
L186
A188
R189
73
R91
F92
L94
T112
L114
W117
P127
N130
A141
A142
—
A152
D158
Y165
A166
G167
L172
T174
A175
74
R92
Y93
V95
T113
L115
W118
W128
P131
S142
H143
—
A153
D159
Y166
A167
S168
L173
V175
E176
75
R92
Y93
V95
T113
L115
W118
W128
P131
S142
H143
—
A153
D159
Y166
A167
S168
L173
V175
E176
76
R89
F90
I92
S110
F112
P115
P125
G128
A140
E141
—
V151
N157
Y164
A165
A166
L171
L173
K174
77
R92
Y93
L95
T113
L115
W118
P128
N131
A142
A143
—
A153
D159
Y166
A167
G168
L173
A175
A176
78
R86
F87
Y89
S107
L109
W112
F122
P125
S136
E137
—
A147
D153
F160
A161
G162
L167
L169
K170
79
R90
F91
F93
S111
V113
F116
Y126
P129
S140
S141
—
A151
D157
F164
A165
G166
L171
V173
K174
80
R93
Y94
Y96
S114
L116
A119
A129
P134
T151
Q152
—
V162
Q168
Y175
A176
G177
L182
V184
R185
81
R92
Y93
V95
T113
L115
W118
W128
P131
S142
H143
—
A153
D159
Y166
A167
S168
L173
V175
E176
82
R87
Y88
V90
T108
L110
L113
P123
K126
A139
S140
—
F150
D156
Y163
A164
G165
L170
L172
E173
83
R89
F90
I92
S110
F112
P115
P125
G128
A140
E141
—
V151
N157
Y164
A165
A166
L171
L173
K174
84
R101
Y102
Y104
S122
L124
V127
L137
P142
R152
Q153
—
V163
E169
Y176
A177
G178
L183
A185
V186
85
R86
Y87
L89
N107
F109
W112
L122
A126
D136
A137
—
I147
N153
Y160
A161
G162
L167
I169
H170
86
R85
Y86
L88
N106
L108
N111
R121
A126
S134
D135
—
Q145
A151
Y158
G159
G160
L165
V167
N168
87
R92
F93
L95
T113
I115
L118
P128
P131
A147
D148
—
F158
D164
Y171
A172
S173
L178
M180
A181
88
C89
R90
I92
T111
L113
L116
P126
R129
G139
A140
—
F150
D156
Y163
A164
G165
L170
T172
R173
89
R84
Y85
F87
N105
F107
W110
L120
T125
A133
D134
—
I144
N150
Y157
A158
G159
L164
M166
E167
90
R93
F94
Y96
S114
L116
W119
P129
S132
A143
S144
—
A154
S160
F167
A168
G169
M174
L176
R177
91
R93
Y94
Y96
S114
L116
V119
L129
A134
R144
Q145
—
V155
E161
Y168
A169
G170
L175
A177
L178
92
R85
F86
Y88
S106
L108
L111
A121
P126
A143
Q144
—
V154
A160
Y167
A168
G169
L174
A176
R177
93
A99
L100
L102
T128
L130
P133
Y143
P146
A162
Q163
—
A173
Q179
Y186
T187
S188
L193
L195
R196
94
R93
F94
Y96
S114
L116
W119
P129
A134
A145
A146
—
A156
A162
F169
A170
G171
M176
L178
V179
95
R87
Y88
L90
G107
L109
W112
P122
S125
A137
D138
—
A148
D154
F161
A162
G163
L168
V170
Q171
96
C88
R89
I91
T110
L112
L115
P125
K128
G138
S139
—
F149
D155
Y162
A163
G164
L169
T171
R172
97
R86
F87
Y89
S107
L109
W112
L122
P125
A135
E136
—
A146
D152
F159
A160
G161
M166
L168
K169
98
R143
F144
L146
F164
L166
I169
A179
P184
E191
S192
R198
V205
E211
Y218
A219
G220
L225
M227
K228
99
R112
Y113
V115
S133
V135
T138
P148
K153
E167
S168
Q174
V181
D187
S194
A195
A196
L201
M203
K204
100
R146
F147
L149
F167
L169
I172
A182
P187
E194
S195
R201
V208
E214
Y221
A222
G223
L228
M230
K231
101
R98
Y99
V101
S119
I121
A124
P134
K139
E152
S153
Q159
V166
D172
S179
A180
G181
L186
M188
C189
102
R142
F143
L145
F163
L165
L168
A178
P183
E190
S191
R197
V204
E210
Y217
A218
G219
L224
M226
K227
103
R102
Y103
V105
S123
L125
T128
P138
K143
E151
S152
Q158
V165
N171
S178
G179
G180
L185
M187
R188
104
R172
F173
L175
F193
L195
F198
A208
P213
E220
S221
R227
V234
E240
Y247
A248
G249
L254
M256
K257
105
R138
F139
L141
F159
L161
I164
A174
P179
E186
S187
R193
V200
E206
Y213
A214
G215
L220
M222
R223
106
R97
Y98
V100
S118
L120
T123
P133
K138
E154
S155
E161
V168
D174
S181
A182
G183
L188
I190
R191
107
R146
F147
L149
F167
L169
I172
A182
P187
E194
S195
R201
V208
E214
Y221
A222
G223
L228
M230
K231
108
R167
F168
L170
F188
L190
I193
A203
P208
E215
S216
R222
V229
E235
Y242
A243
G244
L249
M251
K252
109
R137
F138
L140
F158
L160
V163
A173
P178
E185
S186
R192
V199
E205
Y212
A213
G214
L219
M221
K222
110
R142
Y143
V145
S163
V165
T168
P178
K183
E199
S200
E206
V213
D219
S226
A227
G228
L233
I235
R236
111
R138
F139
L141
F159
L161
I164
A174
P179
E186
S187
R193
V200
E206
Y213
A214
G215
L220
M222
K223
112
R95
Y96
A98
S116
F118
T121
P131
N136
E147
S148
Q154
V161
D167
C174
G175
G176
L181
M183
H184
113
R153
F154
L156
F174
L176
I179
A189
P194
E201
S202
R208
V215
E221
Y228
A229
G230
L235
M237
K238
114
R146
F147
L149
F167
L169
I172
A182
P187
E194
S195
R201
V208
E214
Y221
A222
G223
L228
M230
K231
115
R98
Y99
M101
S119
I121
A124
P134
N139
E152
S153
Q159
V166
D172
S179
A180
G181
L186
M188
C189
116
R167
F168
L170
F188
L190
I193
A203
P208
E215
S216
R222
V229
E235
Y242
A243
G244
L249
M251
K252
117
R99
Y100
V102
S120
V122
T125
P135
N141
E154
S155
Q161
V168
D174
S181
A182
A183
L188
M190
K191
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
ID
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
1
H220
N227
S234
S246
K259
P260
R261
L295
Q301
G308
S324
R335
G346
F349
L351
D352
T358
L384
L397
2
H220
N227
S234
S246
K259
P260
R261
L295
Q301
G308
S324
R335
G346
F349
L351
D352
T358
L384
L397
3
H219
N226
S233
S245
K258
P259
R260
L294
Q300
G307
S323
R334
G345
F348
L350
D351
T357
L383
L396
4
H219
N226
S233
S245
K258
P259
R260
L294
Q300
G307
S323
R334
G345
F348
L350
D351
T357
L383
L396
5
H220
N227
S234
S246
K260
P261
R262
L296
Q302
G309
S325
R336
G347
F350
L352
D353
T359
L385
L398
6
H190
N197
S204
S216
K230
P231
R232
L266
Q272
G279
S295
R306
G317
F320
L322
D323
S329
L355
L368
7
H219
N226
S233
S245
K259
P260
R261
L295
H301
E308
P324
N335
E346
F349
L351
D352
S358
L384
L397
8
H192
N199
S206
A218
K232
K234
R235
L269
H275
E282
S300
C311
G322
F325
L327
D328
S334
L360
L373
9
H191
N198
S205
T217
K231
K233
Q234
F268
P274
E281
S299
C310
R321
F324
L326
N327
S333
L359
L372
10
H190
D197
S204
A216
K229
K231
P232
L266
H272
E279
S297
C308
G319
F322
L324
N325
S331
L357
L370
11
H190
N197
S204
A216
K230
K232
H233
L267
H273
Q280
D294
Y305
G316
F319
L321
N322
S328
L354
L367
12
H185
N192
S199
A211
K225
K227
R228
L262
Q268
E275
S293
C304
G315
F318
L320
D321
V327
L353
L366
13
H190
N197
S204
G216
K230
K232
H233
L267
H273
Q280
D294
G305
G316
F319
L321
N322
T328
L354
L367
14
H187
N194
S201
S213
K227
K229
R230
L264
H270
E277
S295
C306
G317
F320
L322
D323
N329
L355
L368
15
H190
N197
S204
A216
K230
H232
R233
L267
H273
E280
S297
C308
G319
F322
L324
D325
I331
L357
L370
16
H190
N197
S204
A216
K230
H232
R233
L267
H273
E280
S297
C308
G319
F322
L324
D325
I331
L357
L370
17
H190
N197
S204
G216
K230
K232
H233
L267
H273
E280
D294
G305
G316
F319
L321
N322
T328
L354
L367
18
H190
N197
S204
T216
K230
K232
P233
L267
—
Q279
I295
C306
G317
F320
L322
N323
A329
L355
L368
19
H185
N192
S199
T211
K225
K227
P228
L262
—
Q274
I290
C301
G312
F315
L317
N318
A324
L350
L363
20
H188
N195
S202
P214
N228
K230
R231
L265
C271
D278
S296
C307
G318
F321
L323
N324
D330
L356
L369
21
L185
N192
S199
P211
N225
K227
R228
L262
C268
D275
S293
C304
G315
F318
L320
N321
D327
L353
L366
22
H192
N199
S206
A218
T232
K234
G235
L269
—
E280
H298
C309
G320
F323
L325
N326
N332
L358
L371
23
H196
N203
S210
A222
T236
R238
G239
L273
—
E284
H302
C313
G324
F327
L329
N330
N336
L362
L375
24
H192
N199
S203
A215
T229
K231
G232
L266
—
E277
H295
C302
G313
F316
L318
N319
N325
L351
L364
25
H238
N245
S249
A261
T275
K277
G278
L312
—
E323
H341
C348
G359
F362
L364
N365
N371
L397
L410
26
H161
D168
S175
S187
N197
K199
R200
L234
C240
G246
S263
N274
G285
F288
L290
N291
S297
L323
L336
27
H223
N230
S237
A249
K263
R265
N266
L300
F306
G313
T334
S345
G356
F359
L361
D362
N368
L394
L408
28
H225
N232
S239
A251
K265
R267
N268
L302
L308
G315
T336
S347
G358
F361
L363
D364
D370
L396
L410
29
H162
N169
S176
A188
T202
K204
G205
L239
—
E250
H268
C279
G290
F293
L295
N296
N302
L328
L341
30
H225
N232
S239
A251
K265
R267
N268
L302
F308
G315
T336
S347
G358
V361
L363
D364
D370
L396
L410
31
H225
N232
S239
A251
K265
R267
N268
L302
F308
G315
T336
S347
G358
V361
L363
D364
D370
L396
L410
32
H198
N205
S212
D224
K238
R240
—
L274
H280
D287
F308
G319
G330
F333
L335
D336
T342
L368
L381
33
A236
K243
S250
R262
P275
P277
K278
G312
N318
Y325
N335
Y346
—
N357
A359
A360
Y367
L399
L409
34
H192
N199
S206
A218
K232
K234
R235
L269
Y275
E282
S300
C311
R322
F325
L327
D328
S334
L360
L373
35
H279
N286
S293
A305
—
—
—
L331
—
E342
H360
C371
G382
F385
L387
N388
K394
L420
L433
36
H229
D236
S243
A255
K269
R271
N272
L306
C312
D319
L340
S351
G362
F365
L367
D368
D374
L400
L414
37
H189
N196
S203
T215
K229
R231
N232
L266
C272
G279
S300
S311
G322
F325
L327
D328
D334
L360
L374
38
H196
D203
S210
R222
Q236
K237
R238
L272
L278
N285
S306
—
—
—
—
—
T315
L341
L354
39
L188
N195
S202
P214
N228
K230
R231
L265
C271
D278
S296
C307
G318
F321
L323
N324
D330
L356
L369
40
H244
N251
S258
T270
K284
R286
N287
L321
C327
G334
S355
S366
G377
F380
L382
D383
D389
L415
L429
41
H244
N251
S258
T270
K284
R286
N287
L321
C327
G334
S355
S366
G377
F380
L382
D383
D389
L415
L429
42
H189
N196
S203
A215
T229
K231
G232
L266
—
E277
H295
C306
G317
F320
L322
N323
K329
L355
L368
43
H185
S192
S199
K211
T233
P235
R236
T270
Q276
Q283
T303
D314
G325
Y328
L330
D331
I337
L363
L377
44
H185
S192
S199
K211
T233
P235
R236
T270
Q276
Q283
T303
D314
G325
Y328
L330
D331
I337
L363
L377
45
H220
N227
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
46
H219
N226
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
47
H77
N84
S91
A103
K117
R119
N120
L154
F160
G167
T188
S199
G210
V213
L215
D216
D222
L248
L262
48
H106
N113
S120
A132
K146
R148
S149
L183
C189
D196
S217
S228
G239
F242
L244
D245
D251
L277
L291
49
H178
N185
S192
A204
N218
R220
R221
L255
N261
D268
G287
S298
—
—
—
—
—
—
—
50
—
—
—
S3
K17
K19
H20
L54
D60
S67
S88
R99
G110
Y113
L115
D116
M122
L148
L161
51
Y172
E179
S186
—
—
A203
P204
L238
—
P247
F262
H273
P282
D285
—
R286
E293
L319
L330
52
H171
A178
S185
—
—
D204
T205
H238
—
W247
S262
H273
T282
D285
—
L286
T293
L319
L330
53
H171
A178
S185
—
—
D204
T205
H238
—
W247
S262
H273
T282
D285
—
L286
T293
L319
L330
54
—
—
—
—
—
—
—
—
—
—
—
S6
G17
V20
L22
D23
D29
L55
L69
55
Y172
E179
S186
—
—
A203
P204
L238
—
P247
F262
H273
P282
D285
—
R286
E293
L319
L330
56
A170
N177
S184
R196
—
K204
D205
R238
—
—
F261
Y272
S280
K283
R285
A286
L293
L319
L329
57
Q188
E195
S202
R214
—
E219
P220
D254
—
K261
S276
P287
T296
E299
P301
L302
E309
L335
L345
58
H176
A183
S190
R202
—
K210
D211
E244
—
—
S263
H274
D283
D286
D288
M289
I296
L322
L332
59
S173
A180
G187
R199
—
K207
Q208
I241
—
—
V264
Y275
K283
K286
F288
K289
Y296
L322
L332
60
H173
N180
S187
—
G203
F205
F206
I239
S243
—
—
P273
M283
A286
L288
A289
Q295
L321
L331
61
H173
E180
S187
G199
—
A207
T208
T241
—
T250
—
H272
P281
D284
—
T285
P292
L318
L329
62
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
63
S173
R180
G187
R199
—
K207
Q208
L241
—
Q250
T265
Y276
E284
Q287
L289
A290
Y297
L323
L333
64
A180
A187
S194
A206
—
—
A217
D250
I256
G263
H275
H286
—
G297
L299
A300
E307
L333
L343
65
H186
R193
S200
T212
T222
K224
E225
T259
T265
G272
—
H292
A303
K306
F308
K309
K316
L342
L352
66
H173
Q180
S187
Q199
H207
L209
Q210
G244
R250
—
S270
H281
E290
L293
S295
L296
D303
L329
L339
67
F171
A178
G185
R197
—
K205
I206
L239
—
—
R262
Y273
R281
P284
A286
A287
V294
L320
L330
68
Y173
A180
G187
—
G204
K206
A207
E240
Q244
—
—
H270
—
E278
P280
I281
D287
L313
L323
69
A174
E181
S188
K200
G212
K214
M215
T249
—
G257
E272
Y283
—
P294
A296
A297
R304
L330
L340
70
Y171
E178
S185
R197
P209
P211
P212
V246
V252
A257
R283
E294
A303
E306
P308
L309
E316
L342
L352
71
—
—
—
—
—
—
—
T184
C190
L197
—
S213
—
—
—
—
—
—
L230
72
Y190
E197
S204
R216
P228
P230
P231
V265
V271
A276
R299
E310
A319
E322
P324
L325
E332
L358
L368
73
A176
E183
S190
K202
—
K209
M210
V244
—
W252
I267
F278
—
Q289
A291
A292
P299
L325
L335
74
A177
R184
G191
R203
G209
S211
P212
V243
—
G252
T267
P278
—
D289
P291
S292
P299
L325
L335
75
A177
R184
G191
R203
G209
S211
P212
V243
—
G252
T267
P278
—
D289
P291
S292
P299
L325
L335
76
Y175
D182
S189
—
E203
I205
S206
K239
K244
—
V264
H275
I285
S288
L290
L291
Y298
L324
L334
77
A177
E184
S191
K203
—
K210
M211
V245
—
Y253
T268
Y279
—
P290
A292
A293
P300
L326
L336
78
A171
Y178
G185
A197
P206
G208
P209
I241
—
G250
T265
Y276
—
L287
L289
S290
E297
L323
L333
79
A175
H182
G189
K201
T205
G207
P208
V240
—
K249
S264
Y275
—
E286
L288
A289
E296
L322
L332
80
S186
A193
G200
K212
P224
T226
R227
R261
H267
F274
E284
H295
—
S306
I308
A309
P316
L342
L352
81
A177
R184
G191
R203
G209
A211
P212
V243
—
G252
T267
P278
—
E289
P291
S292
P299
L325
L335
82
H174
G181
S188
—
G204
A206
Y207
V240
R244
—
F263
H274
E283
A286
P288
L289
E296
L322
L332
83
Y175
D182
S189
—
E203
I205
S206
K239
K244
—
V264
H275
I285
S288
L290
L291
Y298
L324
L334
84
A187
G194
S201
—
P223
P225
K226
Q260
G265
F272
A282
Y293
—
P304
A306
S307
L314
L340
L350
85
K171
A178
S185
A197
—
—
G198
L232
G238
S245
S253
F264
—
P275
A277
A278
H285
L311
A321
86
K169
E176
S183
G195
—
—
V196
F229
G235
A242
E251
H262
—
P273
M275
S276
N283
L309
S319
87
D182
Q189
S196
Q207
P213
F215
A216
V247
—
G256
R271
A282
—
P293
I295
P296
P303
L329
L339
88
L174
N181
S188
P200
—
R207
A208
L242
G248
K250
I261
Y272
—
K283
Q285
L286
Y293
L318
L328
89
K168
E175
S182
T194
—
—
Q195
E227
G233
—
K245
Y256
—
P267
S269
A270
N277
L303
A313
90
S178
E185
S192
K204
A217
G219
P220
V253
—
—
D274
H285
—
A296
M298
A299
P306
L332
L342
91
A179
G186
S193
R205
P226
P228
K229
H263
E268
F275
A294
Y305
—
P316
A318
S319
P326
L352
L362
92
S178
Q185
G192
R204
P216
V218
R219
E253
N259
F266
E276
Y287
—
P298
A300
S301
P308
L334
L344
93
A197
D204
S211
T223
T229
A231
R232
V266
—
G275
H290
P301
—
P312
L314
S315
E322
V348
F358
94
S180
E187
G194
K206
A219
G221
P222
V255
—
—
D276
Y287
—
G298
M300
S301
P308
L334
L344
95
A172
A179
S186
A194
—
—
—
E236
—
S244
Q255
G266
—
A277
L279
A280
P287
L313
L323
96
L173
D180
S187
P199
—
R206
A207
L241
G247
K249
I260
Y271
—
K282
Q284
L285
Y292
L317
L327
97
A170
R177
G184
A196
P206
G208
P209
V241
—
K250
E265
Y276
—
R287
L289
S290
E297
L323
L333
98
A229
K236
S243
A254
K269
P270
K271
S304
—
G312
—
—
L348
S351
A353
E354
Y360
L386
L402
99
H205
N212
S219
A231
K246
K247
G248
L282
T288
E293
H311
C322
G333
F336
L338
N339
K345
L371
L384
100
A232
K239
S246
E257
K272
P273
K274
S307
—
G315
—
—
L351
A354
A356
D357
Y363
L389
L405
101
H190
D197
S204
A216
R230
K231
A232
L266
H272
E279
S297
C308
G319
F322
L324
N325
S331
L357
L370
102
A228
K235
S242
A253
T268
P269
K270
V303
—
R311
—
—
L347
E350
A352
D353
Y359
L385
L401
103
Y189
D196
S203
S215
S226
K227
R228
L262
C268
G274
P292
G303
G314
F317
L319
N320
S326
L352
L365
104
A258
V265
N272
E283
K298
P299
K300
A333
—
G341
—
—
F377
A380
A382
D383
H389
L415
L431
105
A224
R231
S238
D249
T264
P265
K266
T299
—
Q307
—
—
L343
D346
A348
D349
Y355
L381
L397
106
H192
N199
S206
A218
G233
R234
N235
L269
C275
N282
P303
S314
G325
F328
L330
D331
D337
L363
L377
107
A232
K239
S246
R258
K272
P273
K274
S307
G313
T320
S334
S345
A356
L359
Q361
F362
A368
L395
L405
108
A253
K260
S267
R279
K293
P294
K295
S328
L334
T341
S355
S366
A377
L380
K382
F383
A389
L416
L426
109
A223
R230
S237
K249
A263
P264
K265
T298
D304
G311
S325
S336
A347
L350
K352
F353
A359
L386
L396
110
H237
N244
S251
A263
G278
R279
N280
L314
C320
D327
S348
S359
G370
F373
L375
D376
D382
L408
L422
111
A224
R231
S238
K250
A264
P265
K266
T299
D305
G312
S326
S337
A348
L351
K353
F354
A360
L387
L397
112
L185
N192
S199
P211
K226
K227
R228
L262
C268
D275
S293
C304
G315
F318
L320
N321
D327
L353
L366
113
A239
R246
S253
K265
A279
P280
K281
T314
D320
G327
S341
S352
A363
L366
K368
F369
A375
L402
L412
114
A232
K239
S246
R258
K272
P273
K274
S307
G313
T320
S334
S345
A356
L359
Q361
F362
A368
L395
L405
115
H190
D197
S204
A216
R230
K231
A232
L266
H272
E279
S297
C308
G319
F322
L324
N325
S331
L357
L370
116
A253
K260
S267
R279
K293
P294
K295
S328
L334
T341
S355
S366
A377
L380
K382
F383
A389
L416
L426
117
H192
N199
S206
A218
K233
K234
G235
L269
A275
E280
H298
C309
G320
F323
L325
N326
K332
L358
L371
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
Pos
ID
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
1
F417
T418
T419
F420
A432
T434
K438
L449
T451
F462
Y470
S476
V477
D482
Y493
K498
E515
K528
2
F417
T418
T419
F420
A432
T434
K438
L449
T451
F462
Y470
S476
V477
D482
Y493
K498
E515
K528
3
F416
T417
T418
F419
A431
T433
K437
L448
T450
F461
Y469
C475
V476
D481
Y492
K497
E514
K527
4
F416
T417
T418
F419
A431
T433
K437
L448
T450
F461
Y469
S475
V476
D481
Y492
K497
E514
K527
5
F418
T419
T420
F421
A433
T435
K439
L450
T452
F463
Y471
S477
V478
D483
Y494
K499
E516
K529
6
F388
T389
T390
F391
A403
T405
K409
L420
T422
F433
Y441
S447
V448
D453
Y464
K469
E486
K499
7
Y417
T418
T419
F420
A432
T434
K438
L449
T451
Y462
Y470
S476
V477
E482
Y493
K498
E515
K525
8
Y393
T394
T395
F396
A408
T410
K414
L425
A427
Y438
F446
S452
V453
D458
Y469
K474
D491
K504
9
Y392
T393
T394
F395
A407
T409
K413
L424
A426
Y437
Y445
A451
V452
D457
Y468
K473
D490
K503
10
Y390
T391
T392
F393
A405
T407
K411
L422
V424
Y435
Y443
S449
V450
E455
Y466
K471
D488
K500
11
Y387
T388
T389
F390
A402
T404
R408
L419
A421
Y432
Y440
S446
V447
D452
F463
K468
D485
T498
12
Y386
T387
T388
F389
A401
T403
K407
L418
A420
Y431
Y439
S445
V446
E451
Y462
R467
E484
K497
13
Y387
T388
T389
F390
A402
T404
T408
L419
A421
F432
Y440
S446
V447
D452
Y463
R468
D485
S498
14
Y388
T389
T390
F391
A403
K405
K409
L420
A422
Y433
Y441
S447
V448
E453
Y464
R469
D486
K499
15
Y390
T391
T392
F393
A405
T407
K411
L422
V424
F435
Y443
S449
V450
E455
Y466
K471
E488
K501
16
Y390
T391
T392
F393
A405
T407
K411
L422
V424
F435
Y443
S449
V450
E455
Y466
K471
E488
K501
17
Y387
T388
T389
F390
A402
T404
K408
L419
A421
Y432
Y440
L446
V447
D452
Y463
R468
D485
T498
18
Y388
T389
T390
F391
S403
T405
K409
L420
V422
Y433
Y441
S447
V448
E453
Y464
R469
D486
S499
19
Y383
T384
T385
F386
S398
T400
K404
L415
V417
Y428
Y436
S442
V443
E448
Y459
R464
D481
S494
20
Y389
T390
T391
F392
A404
R406
K410
L421
A423
Y434
Y442
S448
V449
D454
Y465
R470
D487
S500
21
Y386
T387
T388
F389
A401
R403
K407
L418
A420
C431
Y439
S445
V446
D451
Y462
K467
D484
T497
22
Y391
T392
T393
F394
A406
T408
K412
L423
V425
Y436
Y444
S450
V451
D456
Y467
R472
D489
K502
23
Y395
T396
T397
F398
A410
T412
K416
L427
V429
Y440
Y448
S454
V455
D460
Y471
R476
D493
K506
24
Y384
T385
T386
F387
A399
T401
K405
L416
V418
Y429
Y437
S443
V444
D449
Y460
R465
D482
K495
25
Y430
T431
T432
F433
A445
T447
K451
L462
V464
Y475
Y483
S489
V490
D495
Y506
R511
D528
K541
26
Y356
T357
T358
F359
A371
R373
K377
L388
T390
Y401
Y409
S415
V416
E421
Y432
K437
E454
K467
27
Y428
T429
T430
F431
A443
T445
K449
L460
V462
Y473
Y481
S487
V488
E493
Y504
K509
E526
N539
28
Y430
T431
T432
F433
A445
T447
K451
L462
V464
Y475
Y483
S489
V490
E495
Y506
K511
D528
N541
29
Y361
T362
T363
F364
A376
T378
K382
L393
V395
Y406
Y414
S420
V421
D426
Y437
R442
D459
K472
30
Y430
T431
T432
F433
A445
T447
K451
L462
V464
Y475
Y483
S489
V490
E495
Y506
K511
D528
N541
31
Y430
T431
T432
F433
A445
T447
K451
L462
V464
Y475
Y483
S489
V490
E495
Y506
K511
D528
N541
32
Y401
T402
T403
F404
A416
L418
K422
L433
V435
Y446
Y454
L460
V461
D466
Y477
R482
D499
—
33
L429
L430
N431
Y432
K444
E446
V450
L461
K463
V476
F484
D490
L491
T496
L509
V514
A531
F544
34
Y393
T394
T395
F396
—
—
—
L402
C404
Y408
Y416
L422
V423
E428
Y439
R444
D461
K474
35
Y453
T454
T455
F456
A468
T470
K474
L485
V487
Y498
Y506
S512
V513
D518
Y529
R534
D551
K564
36
F434
T435
T436
F437
A449
T451
K455
L466
V468
H479
Y487
L493
A494
G499
Y510
K515
D532
A543
37
Y394
T395
S396
F397
A409
T411
K415
L426
V428
H439
Y447
L453
V454
A459
Y470
K475
D492
D505
38
F374
T375
T376
F377
S389
L391
Q395
L406
V408
Y419
Y427
S433
V434
D439
F450
R455
D472
T485
39
Y389
T390
T391
F392
A404
—
—
—
—
—
—
—
—
—
—
—
—
—
40
Y449
T450
S451
F452
A464
T466
K470
L481
V483
H494
Y502
L508
V509
A514
Y525
—
K537
—
41
Y449
T450
S451
F452
A464
T466
K470
L481
V483
H494
Y502
L508
V509
A514
Y525
—
K537
—
42
Y388
T389
T390
F391
A403
T405
—
—
—
—
—
—
—
—
—
—
—
—
43
F397
T398
T399
F400
R412
K414
L418
I429
V431
Y442
Y450
N456
V457
Q462
Y473
R478
E495
K508
44
F397
T398
T399
F400
R412
K414
L418
I429
V431
Y442
Y450
N456
V457
Q462
Y473
R478
E495
K508
45
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
46
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
47
Y282
T283
T284
F285
A297
T299
K303
L314
V316
Y327
Y335
S341
V342
E347
Y358
K363
D380
N393
48
F311
T312
T313
F314
A326
T328
K332
L343
V345
H356
Y364
S370
A371
G376
—
K383
D400
V413
49
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
50
Y181
T182
T183
F184
A196
L198
R202
L213
V215
Y226
Y234
M240
V241
E246
Y257
S262
—
—
51
L350
T351
T352
F353
L365
E367
E371
L382
I384
R395
Y403
A409
V410
H415
L426
M431
R448
G461
52
L350
T351
T352
F353
S365
A367
Q371
L382
V384
H395
Y403
T409
V410
D415
F426
R431
—
A458
53
L350
T351
T352
F353
S365
A367
Q371
L382
V384
H395
Y403
T409
V410
D415
F426
R431
—
T458
54
Y89
T90
T91
F92
A104
T106
K110
L121
V123
Y134
Y142
S148
V149
E154
Y165
K170
D187
N200
55
L350
T351
T352
F353
L365
E367
E371
L382
I384
R395
Y403
A409
V410
H415
L426
M431
R448
G461
56
F349
T350
T351
F352
K364
D366
K370
V381
L383
R394
Y402
K408
I409
D414
F425
R430
E447
—
57
L365
T366
V367
M368
L380
A382
L386
L397
V399
F410
Y418
H424
F425
A430
M441
R446
E463
—
58
L352
S353
T354
Y355
L367
D369
D373
L384
V386
V397
Y405
K411
I412
N417
F428
R433
Q450
—
59
F352
T353
L354
F355
E367
R369
L373
M384
I386
F397
Y405
E411
H412
D417
I428
R433
N450
—
60
L351
T352
N353
Y354
L366
R368
R372
L383
V385
V396
Y404
E410
Y411
D416
L427
R432
A449
—
61
L349
T350
T351
F352
A364
D366
R370
L381
V383
A394
Y402
A408
A409
E414
F425
R430
R447
V460
62
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
63
F353
T354
L355
F356
M368
K370
I374
M385
I387
L398
Y406
E412
H413
E418
L429
R434
K451
—
64
L363
S364
A365
F366
G378
D380
L384
L395
I397
R408
Y416
E422
L423
S428
L439
R444
E461
—
65
L373
T374
V375
Y376
R388
A390
V394
L405
V407
T418
Y426
E432
S433
A438
F449
R454
E471
—
66
I359
N360
V361
M362
L374
E376
R380
L391
V393
H404
L412
I418
V419
I424
L435
R440
E457
—
67
L350
T351
T352
F353
R365
N367
I371
L382
L384
R395
Y403
E409
I410
D415
F426
R431
E448
—
68
L343
T344
V345
F346
P358
T360
L364
L375
I377
H388
Y396
K402
V403
T408
L419
R424
N441
—
69
T360
I361
N362
L363
Q375
D377
I381
I392
A394
R405
Y413
E419
A420
E425
F436
R441
E458
A471
70
L372
T373
V374
L375
M387
L389
M393
L404
V406
V417
Y425
R431
F432
E437
V448
R453
R470
—
71
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
72
L388
T389
V390
L391
M403
L405
M409
L420
V422
V433
Y441
R447
F448
E453
V464
R469
R486
—
73
T355
T356
T357
L358
R370
D372
I376
L387
I389
R400
Y408
E414
R415
V420
F431
R436
E453
A466
74
L355
T356
V357
F358
A370
D372
E376
L387
V389
R400
L408
E414
T415
R420
L429
L434
E451
—
75
L355
T356
V357
F358
A370
D372
E376
L387
V389
R400
L408
E414
T415
R420
L429
L434
E451
—
76
L354
T355
T356
Y357
L369
Q371
L375
L386
I388
L399
P407
D413
R414
S419
I430
I435
—
—
77
T356
I357
T358
L359
Q371
E373
I377
L388
I390
R401
Y409
E415
R416
E421
L432
R437
E454
A467
78
I353
R354
V355
M356
L368
D370
V374
M385
I387
K398
Y406
E412
R413
F418
L429
Y434
R451
—
79
I352
R353
A354
M355
A367
E369
A373
M384
I386
K397
Y405
E411
K412
F417
L428
Y433
E450
—
80
L372
S373
N374
Y375
M387
D389
V393
L404
A406
L418
Y426
N432
R433
D438
L449
I454
S471
N484
81
L355
T356
V357
F358
T370
D372
E376
L387
V389
R400
F408
E414
T415
R420
L429
L434
E451
—
82
L352
T353
V354
M355
L367
Q369
L373
L384
L386
S397
Y405
P411
W412
K417
F428
V433
E450
—
83
L354
T355
T356
F357
L369
Q371
L375
L386
I388
L399
P407
D413
R414
S419
I430
I435
—
—
84
F370
L371
S372
F373
L394
P396
A400
L411
T413
L425
Y433
Q439
R440
Q445
V457
L462
Q479
—
85
F341
T342
T343
F344
Q356
E358
K362
—
D374
L387
Y395
—
R398
H403
S416
R421
D438
—
86
I339
T340
S341
Y342
L354
E356
L360
—
Q372
L385
F393
—
Y396
H401
I414
Y419
A436
—
87
L359
T360
V361
F362
A374
D376
L380
L391
V393
R404
S412
E418
R419
E424
L433
L438
E455
—
88
Y348
A349
F350
F351
K363
D365
R369
L380
Y382
R395
Y403
S409
R410
D415
I426
K431
E448
—
89
I333
T334
S335
F336
L348
D350
K354
—
K366
R378
Y386
—
D389
H394
V407
E412
E429
—
90
L362
R363
S364
M365
L377
D379
M383
M394
I396
R407
Y415
K421
R422
Q427
L438
Y443
D460
—
91
L382
I383
S384
F385
L406
P408
A412
L423
T425
L437
Y445
Q451
R452
Q457
V469
L474
Q491
—
92
L364
T365
N366
F367
L379
N381
V385
L396
K398
L410
Y418
L424
R425
N430
L441
T446
P463
—
93
L378
R379
T380
F381
H393
D395
N399
L410
V412
R423
Y431
Q437
L438
E443
L454
Y459
E476
—
94
L364
R365
S366
M367
L379
D381
V385
M396
I398
R409
Y417
R423
R424
E429
L440
Y445
D462
—
95
L343
T344
A345
F346
R357
D359
L363
L374
I376
T387
Y395
D401
R402
D407
F418
R423
Q440
—
96
Y347
A348
C349
F350
K362
D364
I368
L379
Y381
R394
Y402
A408
R409
D414
I425
K430
E447
—
97
I353
R354
V355
M356
K368
E370
T374
M385
I387
R398
Y406
E412
K413
F418
F429
Y434
E451
—
98
L422
L423
N424
Y425
K437
E439
V443
L454
S458
V469
F477
D483
L484
K489
L502
V507
T524
—
99
Y404
T405
T406
F407
A419
T421
K425
L436
I438
Y449
Y457
S463
V464
D469
Y480
R485
D502
N515
100
L425
L426
N427
Y428
K440
E442
V446
L457
N461
V472
F480
D486
L487
K492
L505
V510
A527
—
101
Y390
T391
T392
F393
A405
T407
K411
L422
V424
Y435
Y443
S449
V450
E455
Y466
K471
D488
K500
102
L421
L422
N423
Y424
K436
E438
V442
L453
K457
V468
F476
D482
I483
K488
L501
V506
A523
—
103
Y385
T386
T387
F388
A400
R402
K406
L417
A419
Y430
Y438
L444
V445
E450
Y461
K466
E483
K496
104
I451
L452
S453
F454
K466
Q468
A472
L483
S487
V498
F506
D512
L513
K518
L531
V536
A553
—
105
L417
L418
N419
Y420
K432
E434
V438
L449
T453
V464
F472
D478
R479
K484
L497
V502
S519
—
106
F397
T398
T399
F400
A412
T414
K418
L429
V431
H442
Y450
S456
A457
G462
Y473
K478
D495
D508
107
L425
L426
N427
Y428
K440
E442
V446
L457
N461
V472
F480
D486
L487
K492
L505
V510
A527
—
108
I446
L447
S448
Y449
K461
K463
A467
L478
D482
V493
F501
D507
L508
K513
L526
V531
A548
—
109
L416
L417
N418
Y419
K431
E433
V437
L448
R452
V463
F471
D477
R478
K483
L496
V501
S518
—
110
F442
T443
T444
F445
A457
T459
K463
L474
V476
H487
Y495
L501
A502
G507
Y518
K523
D540
G549
111
L417
L418
N419
Y420
K432
E434
V438
L449
R453
V464
F472
D478
R479
K484
L497
V502
S519
—
112
Y386
T387
T388
F389
A401
R403
K407
L418
A420
C431
Y439
S445
V446
D451
Y462
K467
D484
S494
113
L432
L433
N434
Y435
K447
E449
V453
L464
R468
V479
F487
D493
R494
K499
L512
V517
S534
—
114
L425
L426
N427
Y428
K440
E442
V446
L457
N461
V472
F480
D486
L487
K492
L505
V510
A527
—
115
Y390
T391
T392
F393
A405
T407
K411
L422
V424
Y435
Y443
S449
V450
E455
Y466
K471
D488
S497
116
I446
L447
S448
Y449
K461
K463
A467
L478
D482
V493
F501
D507
L508
K513
L526
V531
A548
—
117
Y391
T392
T393
F394
A406
T408
K412
L423
V425
Y436
Y444
S450
V451
D456
Y467
R472
D489
K502
Assays to test for the functionality of such mutants are readily available in the art, and respectively, described in the Example section of the present invention.
In a preferred embodiment, the mutated PPO refers to a polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, or 117, in which the amino acid sequence differs at Mutated site 1 (which corresponds to Arg128 of SEQ ID NO: 1) according to Table 2a, and/or at Mutated site 5 (which corresponds to Phe420 of SEQ ID NO: 1) according to Table 2a.
Examples of differences at these amino acid positions include, but are not limited to, one or more of the following:
the amino acid at Mutated site 1 is other than Arginine (or Tryosine, or Cysteine; as the case may be according to Table 2a);
the amino acid at Mutated site 5 is other than Phenylalanine (or Methionine, or Tyrosine, or Leucine, as the case may be according to Table 2a),
In particularly preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 (which corresponds to Arg128 of SEQ ID NO: 1) is Leu, Ala, Val, Ile, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His, and the amino acid at Mutated site 5 (which corresponds to Phe420 of SEQ ID NO: 1) is Ala, Leu, Val, le, or Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Leu, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Leu, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Leu, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Leu, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Leu, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ala, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ala, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ala, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ala, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ala, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Val, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Val, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Val, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Val, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Val, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ile, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ile, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ile, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ile, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ile, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Met, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Met, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Met, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Met, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Met, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Tyr, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Tyr, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Tyr, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Tyr, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Tyr, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gly, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gly, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gly, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gly, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gly, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Asn, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Asn, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Asn, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Asn, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Asn, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Cys, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Cys, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Cys, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Cys, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Cys, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Phe, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 1 is Phe, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Phe, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Phe, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Phe, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ser, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ser, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ser, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ser, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Ser, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Thr, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Thr, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Thr, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Thr, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Thr, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gln, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gln, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gln, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gln, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is Gln, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is His, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is His, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is His, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is His, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 1 is His, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO refers to a polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116,or 117, in which the amino acid sequence differs at Mutated site 4 (which corresponds to Leu397 of SEQ ID NO: 1) according to Table 2a, and/or at Mutated site 5 (which corresponds to Phe420 of SEQ ID NO: 1) according to Table 2a.
Examples of differences at these amino acid positions include, but are not limited to, one or more of the following:
the amino acid at Mutated site 4 is other than Leucine (or Alanine, or Serine, or Phenylalanine, as the case may be according to Table 2a);
the amino acid at Mutated site 5 is other than Phenylalanine (or Methionine, or Tyrosine, or Leucine, as the case may be according to Table 2a),
In particularly preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 (which corresponds to Leu397 of SEQ ID NO: 1) is Ala, Arg, Val, Ile, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Gly, Pro, Phe, Tyr, or Trp, and the amino acid at Mutated site 5 (which corresponds to Phe420 of SEQ ID NO: 1) is Ala, Leu, Val, lie, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Gly, Pro, Arg, Tyr, or Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ala, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Arg, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Val, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ile, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ile, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ile, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ile, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is lie, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Met, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is His, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Lys, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asp, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Glu, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Ser, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Thr, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Asn, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gln, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Cys, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Gly, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which: the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Pro, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Phe, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Tyr, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Leu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Val.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Ile.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Met.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is His.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Lys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Asp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Glu.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Ser.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Thr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Asn.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Gln.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Cys.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Gly.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Pro.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Tyr.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 4 is Trp, and the amino acid at Mutated site 5 is Trp.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 3 is other than Gly, is preferably Arg.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 6 is other than Gly, is preferably Ala.
In another preferred embodiment, the mutated PPO comprises a sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, a variant, derivative, orthologue, paralogue or homologue thereof, in which:
the amino acid at Mutated site 7 is other than Leu, is preferably Phe.
It will be within the knowledge of the skilled artisan to identify conserved regions and motifs shared between the homologues, orthologues and paralogues of PPO polypeptides comprising SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637. Having identified such conserved regions that may represent suitable binding motifs, amino acids corresponding to the amino acids listed in Table 2a and 2b can be chosen to be substituted by any other amino acid, preferably by the amino acids listed under mutated sites 1, 2, 3, 4, or 5.
Accordingly, preferred motifs shared between the homologues, orthologues and paralogues of PPO polypeptides described above are Motif 1: SQ[N/K/H]KRYI, wherein the Arg at position 5 within said motif is substituted by Leu, Ala, Val, lie, Met, Tyr, Gly, Asn, Cys, Phe, Ser, Thr, Gln, or His; Motif 2: TLGTLFSS, wherein the Leu at position 2 within said motif is substituted by Ala, Arg, Val, lie, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Gly, Pro, Phe, Tyr, or Trp; and/or wherein the Gly at position 3 within said motif is substituted by Ala, Arg, Val, Ile, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Leu, Pro, Phe, Tyr, or Trp; and/or wherein the Leu at position 5 within said motif is substituted by Ala, Arg, Val, le, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Gly, Pro, Phe, Tyr, or Trp; Motif 3: [F/Y]TTF[V/I]GG, wherein the Phe at position 4 within said motif is substituted by Ala, Leu, Val, lie, Met, His, Lys, Asp, Glu, Ser, Thr, Asn, Gln, Cys, Gly, Pro, Arg, Tyr, or Trp.
Further examples of such homologues, orthologues and paralogues are PPO polypeptides comprising the amino acid sequence of SEQ ID NO: 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637.
In addition, the present invention refers to a method for identifying a uracilpyridine herbicide by using a herbicide tolerant PPO polypeptide as defined SUPRA.
a) generating a transgenic cell or plant comprising a nucleic acid encoding a mutated PPO, wherein the mutated PPO is expressed;
b) applying a uracilpyridine herbicide to the transgenic cell or plant of a) and to a control cell or plant of the same variety;
c) determining the growth or the viability of the transgenic cell or plant and the control cell or plant after application of said uracilpyridine herbicide, and
d) selecting “uracilpyridine herbicides” which confer reduced growth to the control cell or plant as compared to the growth of the transgenic cell or plant.
Said method comprises the steps of:
By “control cell” or “similar, wild-type, plant, plant tissue, plant cell or host cell” is intended a plant, plant tissue, plant cell, or host cell, respectively, that lacks the herbicide-resistance characteristics and/or particular polynucleotide of the invention that are disclosed herein. The use of the term “wild-type” is not, therefore, intended to imply that a plant, plant tissue, plant cell, or other host cell lacks recombinant DNA in its genome, and/or does not possess herbicide-resistant characteristics that are different from those disclosed herein.
a) generating a library of mutated PPO-encoding nucleic acids,
b) screening a population of the resulting mutated PPO-encoding nucleic acids by expressing each of said nucleic acids in a cell or plant and treating said cell or plant with a uracilpyridine herbicide,
c) comparing the uracilpyridine herbicide-tolerance levels provided by said population of mutated PPO encoding nucleic acids with the uracilpyridine herbicide-tolerance level provided by a control PPO-encoding nucleic acid,
d) selecting at least one mutated PPO-encoding nucleic acid that provides a significantly increased level of tolerance to a uracilpyridine herbicide as compared to that provided by the control PPO-encoding nucleic acid.
Another object refers to a method of identifying a nucleotide sequence encoding a mutated PPO which is resistant or tolerant to a uracilpyridine herbicide, the method comprising:
In a preferred embodiment, the mutated PPO-encoding nucleic acid selected in step d) provides at least 2-fold as much resistance or tolerance of a cell or plant to a uracilpyridine herbicide as compared to that provided by the control PPO-encoding nucleic acid.
In a further preferred embodiment, the mutated PPO-encoding nucleic acid selected in step d) provides at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, at least 500-fold, as much resistance or tolerance of a cell or plant to a uracilpyridine herbicide as compared to that provided by the control PPO-encoding nucleic acid.
The resistance or tolerance can be determined by generating a transgenic plant or host cell, preferably a plant cell, comprising a nucleic acid sequence of the library of step a) and comparing said transgenic plant with a control plant or host cell, preferably a plant cell.
a) identifying an effective amount of a uracilpyridine herbicide in a culture of plant cells or green algae that leads to death of said cells.
b) treating said plant cells or green algae with a mutagenizing agent,
c) contacting said mutagenized cells population with an effective amount of uracilpyridine herbicide, identified in a),
d) selecting at least one cell surviving these test conditions,
e) PCR-amplification and sequencing of PPO genes from cells selected in d) and comparing such sequences to wild-type PPO gene sequences, respectively.
Another object refers to a method of identifying a plant or algae containing a nucleic acid comprising a nucleotide sequence encoding a wild-type or mutated PPO which is resistant or tolerant to a uracilpyridine herbicide, the method comprising:
In a preferred embodiment, said mutagenizing agent is ethylmethanesulfonate (EMS). Many methods well known to the skilled artisan are available for obtaining suitable candidate nucleic acids for identifying a nucleotide sequence encoding a mutated PPO from a variety of different potential source organisms including microbes, plants, fungi, algae, mixed cultures etc. as well as environmental sources of DNA such as soil. These methods include inter alia the preparation of cDNA or genomic DNA libraries, the use of suitably degenerate oligonucleotide primers, the use of probes based upon known sequences or complementation assays (for example, for growth upon tyrosine) as well as the use of mutagenesis and shuffling in order to provide recombined or shuffled mutated PPO-encoding sequences.
Arabidopsis
E. coli
Nucleic acids comprising candidate and control PPO encoding sequences can be expressed in yeast, in a bacterial host strain, in an alga or in a higher plant such as tobacco or and the relative levels of inherent tolerance of the PPO encoding sequences screened according to a visible indicator phenotype of the transformed strain or plant in the presence of different concentrations of the selected uracilpyridine herbicide. Dose responses and relative shifts in dose responses associated with these indicator phenotypes (formation of brown color, growth inhibition, herbicidal effect etc) are conveniently expressed in terms, for example, of GR50 (concentration for 50% reduction of growth) or MIC (minimum inhibitory concentration) values where increases in values correspond to increases in inherent tolerance of the expressed PPO. For example, in a relatively rapid assay system based upon transformation of a bacterium such as , each mutated PPO encoding sequence may be expressed, for example, as a DNA sequence under expression control of a controllable promoter such as the lacZ promoter and taking suitable account, for example by the use of synthetic DNA, of such issues as codon usage in order to obtain as comparable a level of expression as possible of different PPO sequences. Such strains expressing nucleic acids comprising alternative candidate PPO sequences may be plated out on different concentrations of the selected uracilpyridine herbicide in, optionally, a tyrosine supplemented medium and the relative levels of inherent tolerance of the expressed PPO enzymes estimated on the basis of the extent and MIC for inhibition of the formation of the brown, ochronotic pigment.
Agrobacterium
Arabidopsis
In another embodiment, candidate nucleic acids are transformed into plant material to generate a transgenic plant, regenerated into morphologically normal fertile plants which are then measured for differential tolerance to selected uracilpyridine herbicides as described in the Example section hereinafter. Many suitable methods for transformation using suitable selection markers such as kanamycin, binary vectors such as from and plant regeneration as, for example, from tobacco leaf discs are well known in the art. Optionally, a control population of plants is likewise transformed with a nucleic acid expressing the control PPO. Alternatively, an untransformed dicot plant such as or Tobacco can be used as a control since this, in any case, expresses its own endogenous PPO. The average, and distribution, of herbicide tolerance levels of a range of primary plant transformation events or their progeny to uracilpyridine herbicides described supra are evaluated in the normal manner based upon plant damage, meristematic bleaching symptoms etc. at a range of different concentrations of herbicides. These data can be expressed in terms of, for example, GR50 values derived from dose/response curves having “dose” plotted on the x-axis and “percentage kill”, “herbicidal effect”, “numbers of emerging green plants” etc. plotted on the y-axis where increased GR50 values correspond to increased levels of inherent tolerance of the expressed PPO. Herbicides can suitably be applied pre-emergence or post-emergence.
In another embodiment, the invention refers to a plant cell transformed by a nucleic acid encoding a herbicide tolerant PPO polypeptide disclosed herein or to a plant cell which has been mutated to obtain a plant expressing a nucleic acid encoding a mutated PPO polypeptide according to the present invention, wherein expression of the nucleic acid in the plant cell results in increased resistance or tolerance to a uracilpyridine herbicide as compared to a wild type variety of the plant cell.
The term “expression/expressing” or “gene expression” means the transcription of a specific gene or specific genes or specific genetic construct. The term “expression” or “gene expression” in particular means the transcription of a gene or genes or genetic construct into structural RNA (rRNA, tRNA) or mRNA with or without subsequent translation of the latter into a protein. The process includes transcription of DNA and processing of the resulting mRNA product.
To obtain the desired effect, i.e. plants that are tolerant or resistant to the uracilpyridine herbicide derivative herbicide of the present invention, it will be understood that the at least one nucleic acid is “over-expressed” by methods and means known to the person skilled in the art.
The term “increased expression” or “overexpression” as used herein means any form of expression that is additional to the original wild-type expression level. Methods for increasing expression of genes or gene products are well documented in the art and include, for example, overexpression driven by appropriate promoters, the use of transcription enhancers or translation enhancers. Isolated nucleic acids which serve as promoter or enhancer elements may be introduced in an appropriate position (typically upstream) of a non-heterologous form of a polynucleotide so as to upregulate expression of a nucleic acid encoding the polypeptide of interest. For example, endogenous promoters may be altered in vivo by mutation, deletion, and/or substitution (see, Kmiec, U.S. Pat. No. 5,565,350; Zarling et al., WO9322443), or isolated promoters may be introduced into a plant cell in the proper orientation and distance from a gene of the present invention so as to control the expression of the gene.
If polypeptide expression is desired, it is generally desirable to include a polyadenylation region at the 3′-end of a polynucleotide coding region. The polyadenylation region can be derived from the natural gene, from a variety of other plant genes, or from T-DNA. The 3′ end sequence to be added may be derived from, for example, the nopaline synthase or octopine synthase genes, or alternatively from another plant gene, or less preferably from any other eukaryotic gene.
An intron sequence may also be added to the 5′ untranslated region (UTR) or the coding sequence of the partial coding sequence to increase the amount of the mature message that accumulates in the cytosol. Inclusion of a spliceable intron in the transcription unit in both plant and animal expression constructs has been shown to increase gene expression at both the mRNA and protein levels up to 1000-fold (Buchman and Berg (1988) Mol. Cell biol. 8: 4395-4405; Callis et al. (1987) Genes Dev 1:1183-1200). Such intron enhancement of gene expression is typically greatest when placed near the 5′ end of the transcription unit. Use of the maize introns Adh1-S intron 1, 2, and 6, the Bronze-1 intron are known in the art. For general information see: The Maize Handbook, Chapter 116, Freeling and Walbot, Eds., Springer, N.Y. (1994)
The term “introduction” or “transformation” as referred to herein encompasses the transfer of an exogenous polynucleotide into a host cell, irrespective of the method used for transfer. Plant tissue capable of subsequent clonal propagation, whether by organogenesis or embryogenesis, may be transformed with a genetic construct of the present invention and a whole plant regenerated there from. The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed. Exemplary tissue targets include leaf disks, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing meristematic tissue (e.g., apical meristem, axillary buds, and root meristems), and induced meristem tissue (e.g., cotyledon meristem and hypocotyl meristem). The polynucleotide may be transiently or stably introduced into a host cell and may be maintained non-integrated, for example, as a plasmid. Alternatively, it may be integrated into the host genome. The resulting transformed plant cell may then be used to regenerate a transformed plant in a manner known to persons skilled in the art.
The transfer of foreign genes into the genome of a plant is called transformation. Transformation of plant species is now a fairly routine technique. Advantageously, any of several transformation methods may be used to introduce the gene of interest into a suitable ancestor cell. The methods described for the transformation and regeneration of plants from plant tissues or plant cells may be utilized for transient or for stable transformation. Transformation methods include the use of liposomes, electroporation, chemicals that increase free DNA uptake, injection of the DNA directly into the plant, particle gun bombardment, transforrmation using viruses or pollen and microprojection.
Agrobacterium
Agrobacterium
Agrobacterium tumefaciens
Arabidopsis
Arabidopsis thaliana
Agrobacterium tumefaciens
Methods may be selected from the calcium/polyethylene glycol method for protoplasts (Krens, F. A. et al., (1982) Nature 296, 72-74; Negrutiu I et al. (1987) Plant Mol Biol 8: 363-373); electroporation of protoplasts (Shillito R. D. et al. (1985) Bio/Technol 3, 1099-1102); microinjection into plant material (Crossway A et al., (1986) Mol. Gen Genet 202: 179-185); DNA or RNA-coated particle bombardment (Klein T M et al., (1987) Nature 327: 70) infection with (non-integrative) viruses and the like. Transgenic plants, including transgenic crop plants, are preferably produced via -mediated transformation. An advantageous transformation method is the transformation in planta. To this end, it is possible, for example, to allow the agrobacteria to act on plant seeds or to inoculate the plant meristem with agrobacteria. It has proved particularly expedient in accordance with the invention to allow a suspension of transformed agrobacteria to act on the intact plant or at least on the flower primordia. The plant is subsequently grown on until the seeds of the treated plant are obtained (Clough and Bent, Plant J. (1998) 16, 735-743). Methods for -mediated transformation of rice include well known methods for rice transformation, such as those described in any of the following: European patent application EP 1198985 A1, Aldemita and Hodges (Planta 199: 612-617, 1996); Chan et al. (Plant Mol Biol 22 (3): 491-506, 1993), Hiei et al. (Plant J 6 (2): 271-282, 1994), which disclosures are incorporated by reference herein as if fully set forth. In the case of corn transformation, the preferred method is as described in either Ishida et al. (Nat. Biotechnol 14(6): 745-50, 1996) or Frame et al. (Plant Physiol 129(1): 13-22, 2002), which disclosures are incorporated by reference herein as if fully set forth. Said methods are further described by way of example in B. Jenes et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1, Engineering and Utilization, eds. S. D. Kung and R. Wu, Academic Press (1993) 128-143 and in Potrykus Annu. Rev. Plant Physiol. Plant Molec. Biol. 42 (1991) 205-225). The nucleic acids or the construct to be expressed is preferably cloned into a vector, which is suitable for transforming , for example pBin19 (Bevan et al., Nucl. Acids Res. 12 (1984) 8711). Agrobacteria transformed by such a vector can then be used in known manner for the transformation of plants, such as plants used as a model, like (is within the scope of the present invention not considered as a crop plant), or crop plants such as, by way of example, tobacco plants, for example by immersing bruised leaves or chopped leaves in an agrobacterial solution and then culturing them in suitable media. The transformation of plants by means of is described, for example, by Höfgen and Willmitzer in Nucl. Acid Res. (1988) 16, 9877 or is known inter ala from F. F. White, Vectors for Gene Transfer in Higher Plants; in Transgenic Plants, Vol. 1, Engineering and Utilization, eds. S. D. Kung and R. Wu, Academic Press, 1993, pp. 15-38.
Arabidopsis
Arabidopsis
Arabidopsis
In addition to the transformation of somatic cells, which then have to be regenerated into intact plants, it is also possible to transform the cells of plant meristems and in particular those cells which develop into gametes. In this case, the transformed gametes follow the natural plant development, giving rise to transgenic plants. Thus, for example, seeds of are treated with agrobacteria and seeds are obtained from the developing plants of which a certain proportion is transformed and thus transgenic [Feldman, K A and Marks M D (1987). Mol Gen Genet 208:274-289; Feldmann K (1992). In: C Koncz, N-H Chua and J Shell, eds, Methods in Research. Word Scientific, Singapore, pp. 274-289]. Alternative methods are based on the repeated removal of the inflorescences and incubation of the excision site in the center of the rosette with transformed agrobacteria, whereby transformed seeds can likewise be obtained at a later point in time (Chang (1994). Plant J. 5: 551-558; Katavic (1994). Mol Gen Genet, 245: 363-370). However, an especially effective method is the vacuum infiltration method with its modifications such as the “floral dip” method. In the case of vacuum infiltration of , intact plants under reduced pressure are treated with an agrobacterial suspension [Bechthold, N (1993). C R Acad Sci Paris Life Sci, 316: 1194-1199], while in the case of the “floral dip” method the developing floral tissue is incubated briefly with a surfactant-treated agrobacterial suspension [Clough, S J and Bent A F (1998) The Plant J. 16, 735-743]. A certain proportion of transgenic seeds are harvested in both cases, and these seeds can be distinguished from non-transgenic seeds by growing under the above-described selective conditions. In addition the stable transformation of plastids is of advantages because plastids are inherited maternally is most crops reducing or eliminating the risk of transgene flow through pollen. The transformation of the chloroplast genome is generally achieved by a process which has been schematically displayed in Klaus et al., 2004 [Nature Biotechnology 22 (2), 225-229]. Briefly the sequences to be transformed are cloned together with a selectable marker gene between flanking sequences homologous to the chloroplast genome. These homologous flanking sequences direct site specific integration into the plastome. Plastidal transformation has been described for many different plant species and an overview is given in Bock (2001) Transgenic plastids in basic research and plant biotechnology. J Mol Biol. 2001 Sep. 21; 312 (3):425-38 or Maliga, P (2003) Progress towards commercialization of plastid transformation technology. Trends Biotechnol. 21, 20-28. Further biotechnological progress has recently been reported in form of marker free plastid transformants, which can be produced by a transient co-integrated maker gene (Klaus et al., 2004, Nature Biotechnology 22(2), 225-229). The genetically modified plant cells can be regenerated via all methods with which the skilled worker is familiar. Suitable methods can be found in the abovementioned publications by S. D. Kung and R. Wu, Potrykus or Höfgen and Willmitzer.
Generally after transformation, plant cells or cell groupings are selected for the presence of one or more markers which are encoded by plant-expressible genes co-transferred with the gene of interest, following which the transformed material is regenerated into a whole plant. To select transformed plants, the plant material obtained in the transformation is, as a rule, subjected to selective conditions so that transformed plants can be distinguished from untransformed plants. For example, the seeds obtained in the above-described manner can be planted and, after an initial growing period, subjected to a suitable selection by spraying. A further possibility consists in growing the seeds, if appropriate after sterilization, on agar plates using a suitable selection agent so that only the transformed seeds can grow into plants. Alternatively, the transformed plants are screened for the presence of a selectable marker such as the ones described above.
Following DNA transfer and regeneration, putatively transformed plants may also be evaluated, for instance using Southern analysis, for the presence of the gene of interest, copy number and/or genomic organisation. Alternatively or additionally, expression levels of the newly introduced DNA may be monitored using Northern and/or Western analysis, both techniques being well known to persons having ordinary skill in the art.
The generated transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, a first generation (or T1) transformed plant may be selfed and homozygous second-generation (or T2) transformants selected, and the T2 plants may then further be propagated through classical breeding techniques. The generated transformed organisms may take a variety of forms. For example, they may be chimeras of transformed cells and non-transformed cells; clonal transformants (e.g., all cells transformed to contain the expression cassette); grafts of transformed and untransformed tissues (e.g., in plants, a transformed rootstock grafted to an untransformed scion).
Preferably, the wild-type or mutated PPO nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide encoding a polypeptide as shown in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637,or a variant or derivative thereof; b) a polynucleotide comprising at least 60 consecutive nucleotides of any of a); and c) a polynucleotide complementary to the polynucleotide of any of a) through b).
Preferably, the expression of the nucleic acid in the plant results in the plant's increased resistance to uracilpyridine herbicide as compared to a wild type variety of the plant.
In another embodiment, the invention refers to a plant, preferably a transgenic plant, comprising a plant cell according to the present invention, wherein expression of the nucleic acid in the plant results in the plant's increased resistance to uracilpyridine herbicide as compared to a wild type variety of the plant.
(a) the nucleic acid sequences encoding proteins useful in the methods of the invention, or
(b) genetic control sequence(s) which is operably linked with the nucleic acid sequence according to the invention, for example a promoter, or
(c) a) and b)
The plants described herein can be either transgenic crop plants or non-transgenic plants. For the purposes of the invention, “transgenic”, “transgene” or “recombinant” means with regard to, for example, a nucleic acid sequence, an expression cassette, gene construct or a vector comprising the nucleic acid sequence or an organism transformed with the nucleic acid sequences, expression cassettes or vectors according to the invention, all those constructions brought about by recombinant methods in which either
are not located in their natural genetic environment or have been modified by recombinant methods, it being possible for the modification to take the form of, for example, a substitution, addition, deletion, inversion or insertion of one or more nucleotide residues in order to allow for the expression of the mutated PPO of the present invention. The natural genetic environment is understood as meaning the natural genomic or chromosomal locus in the original plant or the presence in a genomic library. In the case of a genomic library, the natural genetic environment of the nucleic acid sequence is preferably retained, at least in part. The environment flanks the nucleic acid sequence at least on one side and has a sequence length of at least 50 bp, preferably at least 500 bp, especially preferably at least 1000 bp, most preferably at least 5000 bp. A naturally occurring expression cassette—for example the naturally occurring combination of the natural promoter of the nucleic acid sequences with the corresponding nucleic acid sequence encoding a polypeptide useful in the methods of the present invention, as defined above—becomes a transgenic expression cassette when this expression cassette is modified by non-natural, synthetic (“artificial”) methods such as, for example, mutagenic treatment. Suitable methods are described, for example, in U.S. Pat. No. 5,565,350 or WO 00/15815.
A transgenic plant for the purposes of the invention is thus understood as meaning, as above, that the nucleic acids of the invention are not at their natural locus in the genome of said plant, it being possible for the nucleic acids to be expressed homologously or heterologously. However, as mentioned, transgenic also means that, while the nucleic acids according to the invention or used in the inventive method are at their natural position in the genome of a plant, the sequence has been modified with regard to the natural sequence, and/or that the regulatory sequences of the natural sequences have been modified. Transgenic is preferably understood as meaning the expression of the nucleic acids according to the invention at an unnatural locus in the genome, i.e. homologous or, preferably, heterologous expression of the nucleic acids takes place. Preferred transgenic plants are mentioned herein. Furthermore, the term “transgenic” refers to any plant, plant cell, callus, plant tissue, or plant part, that contains all or part of at least one recombinant polynucleotide. In many cases, all or part of the recombinant polynucleotide is stably integrated into a chromosome or stable extra-chromosomal element, so that it is passed on to successive generations. For the purposes of the invention, the term “recombinant polynucleotide” refers to a polynucleotide that has been altered, rearranged, or modified by genetic engineering. Examples include any cloned polynucleotide, or polynucleotides, that are linked or joined to heterologous sequences. The term “recombinant” does not refer to alterations of polynucleotides that result from naturally occurring events, such as spontaneous mutations, or from non-spontaneous mutagenesis followed by selective breeding.
Plants containing mutations arising due to non-spontaneous mutagenesis and selective breeding are referred to herein as non-transgenic plants and are included in the present invention. In embodiments wherein the plant is transgenic and comprises multiple mutated PPO nucleic acids, the nucleic acids can be derived from different genomes or from the same genome. Alternatively, in embodiments wherein the plant is non-transgenic and comprises multiple mutated PPO nucleic acids, the nucleic acids are located on different genomes or on the same genome.
As used herein, “mutagenized” refers to an organism or DNA thereof having alteration(s) in the biomolecular sequence of its native genetic material as compared to the sequence of the genetic material of a corresponding wild-type organism or DNA, wherein the alteration(s) in genetic material were induced and/or selected by human action. Methods of inducing mutations can induce mutations in random positions in the genetic material or can induce mutations in specific locations in the genetic material (i.e., can be directed mutagenesis techniques), such as by use of a genoplasty technique.
In certain embodiments, the present invention involves herbicide-resistant plants that are produced by mutation breeding. Such plants comprise a polynucleotide encoding a mutated PPO and are tolerant to one or more uracilpyridine herbicides. Such methods can involve, for example, exposing the plants or seeds to a mutagen, particularly a chemical mutagen such as, for example, ethyl methanesulfonate (EMS) and selecting for plants that have enhanced tolerance to at least one or more uracilpyridine herbicide.
However, the present invention is not limited to herbicide-tolerant plants that are produced by a mutagenesis method involving the chemical mutagen EMS. Any mutagenesis method known in the art may be used to produce the herbicide-resistant plants of the present invention. Such mutagenesis methods can involve, for example, the use of any one or more of the following mutagens: radiation, such as X-rays, Gamma rays (e.g., cobalt 60 or cesium 137), neutrons, (e.g., product of nuclear fission by uranium 235 in an atomic reactor), Beta radiation (e.g., emitted from radioisotopes such as phosphorus 32 or carbon 14), and ultraviolet radiation (preferably from 2500 to 2900 nm), and chemical mutagens such as base analogues (e.g., 5-bromo-uracil), related compounds (e.g., 8-ethoxy caffeine), antibiotics (e.g., streptonigrin), alkylating agents (e.g., sulfur mustards, nitrogen mustards, epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, or acridines. Herbicide-resistant plants can also be produced by using tissue culture methods to select for plant cells comprising herbicide-resistance mutations and then regenerating herbicide-resistant plants therefrom. See, for example, U.S. Pat. Nos. 5,773,702 and 5,859,348, both of which are herein incorporated in their entirety by reference. Further details of mutation breeding can be found in “Principals of Cultivar Development” Fehr, 1993 Macmillan Publishing Company the disclosure of which is incorporated herein by reference
Genome
scale engineering for systems and synthetic biology”, Mol Syst Biol.
Precision editing of large animal genomes” Adv Genet.
Gene targeting in plants:
years later”, Int. J. Dev. Biol.
Rewriting the book of life: a new era in precision genome editing”, Boston Consulting Group
Alternatively, herbicide-resistant plants according to the present invention can also be produced by using genome editing methods to select for plant cells comprising herbicide-resistance mutations and then regenerating herbicide-resistant plants therefrom. “Genome Editing” refers to a type of genetic engineering in which DNA is inserted, deleted or replaced in the genome of an organism using engineered nucleases. These nucleases are known to the skilled artisan to create site-specific double-strand breaks at desired locations in the genome. The induced double-strand breaks are repaired through nonhomologous end-joining or homologous recombination, resulting in targeted mutations. Known in the art are currently four families of engineered nucleases which can be used for the purposes of the present invention: meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector-based nucleases (TALEN), and the CRISPR-Cas system. For references, see, for example, Esvelt, K M. and Wang, H H. (2013) “-9 (1): 641; Tan, W S. et al., (2012) “80: 37-97; Puchta, H. and Fauser, F. (2013) “25 57 629-637 Boglioli, Elsy and Richard, Magali “, Retrieved Nov. 30, 2015; Method of the Year 2011. Nat Meth 9 (1), 1-1.
Consequently, in another embodiment, the invention refers to a non-transgenic plant, comprising a plant cell according to the present invention, wherein expression of the nucleic acid encoding a mutated PPO in the plant results in the plant's increased resistance to uracilpyridine herbicide as compared to a wild type variety of the plant.
In addition to the definition above, the term “plant” is intended to encompass crop plants at any stage of maturity or development, as well as any tissues or organs (plant parts) taken or derived from any such plant unless otherwise clearly indicated by context. Plant parts include, but are not limited to, stems, roots, flowers, ovules, stamens, leaves, embryos, meristematic regions, callus tissue, anther cultures, gametophytes, sporophytes, pollen, microspores, protoplasts, and the like.
The plant of the present invention comprises at least one mutated PPO nucleic acid or over-expressed wild-type PPO nucleic acid, and has increased tolerance to a uracilpyridine herbicide as compared to a wild-type variety of the plant. It is possible for the plants of the present invention to have multiple wild-type or mutated PPO nucleic acids from different genomes since these plants can contain more than one genome. For example, a plant contains two genomes, usually referred to as the A and B genomes. Because PPO is a required metabolic enzyme, it is assumed that each genome has at least one gene coding for the PPO enzyme (i.e. at least one PPO gene). As used herein, the term “PPO gene locus” refers to the position of an PPO gene on a genome, and the terms “PPO gene” and “PPO nucleic acid” refer to a nucleic acid encoding the PPO enzyme. The PPO nucleic acid on each genome differs in its nucleotide sequence from an PPO nucleic acid on another genome. One of skill in the art can determine the genome of origin of each PPO nucleic acid through genetic crossing and/or either sequencing methods or exonuclease digestion methods known to those of skill in the art.
The present invention includes plants comprising one, two, three, or more mutated PPO alleles, wherein the plant has increased tolerance to a uracilpyridine herbicide as compared to a wild-type variety of the plant. The mutated PPO alleles can comprise a nucleotide sequence selected from the group consisting of a polynucleotide encoding a polypeptide as defined in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565,or a variant or derivative, homologue, orthologue, paralogue thereof, a polynucleotide comprising at least 60 consecutive nucleotides of any of the aforementioned polynucleotides; and a polynucleotide complementary to any of the aforementioned polynucleotides.
Oryza sativa
In a particularly preferred embodiment, the mutagenized plant refers to a rice plant of the species which comprises a mutated PPO allele which comprises a polynucleotide encoding a polypeptide as defined in SEQ ID NO: 628, 629, 630, 631, 632, 633, 634, 635, or 636.
Preferably, the polynucleotide comprises a nucleic acid sequence as defined in SEQ ID NO: 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648,or 649.
Said mutated rice lines were deposited under the terms of the Budapest Treaty on Nov. 8, 2017, and have been assigned the NCIMB accession numbers 42873, 42876, 42878, 42871, 42874, 42875, 42870, 42877,and 42872.
“Alleles” or “allelic variants” are alternative forms of a given gene, located at the same chromosomal position. Allelic variants encompass Single Nucleotide Polymorphisms (SNPs), as well as Small Insertion/Deletion Polymorphisms (INDELs). The size of INDELs is usually less than 100 bp. SNPs and INDELs form the largest set of sequence variants in naturally occurring polymorphic strains of most organisms
The term “variety” refers to a group of plants within a species defined by the sharing of a common set of characteristics or traits accepted by those skilled in the art as sufficient to distinguish one cultivar or variety from another cultivar or variety. There is no implication in either term that all plants of any given cultivar or variety will be genetically identical at either the whole gene or molecular level or that any given plant will be homozygous at all loci. A cultivar or variety is considered “true breeding” for a particular trait if, when the true-breeding cultivar or variety is self-pollinated, all of the progeny contain the trait. The terms “breeding line” or “line” refer to a group of plants within a cultivar defined by the sharing of a common set of characteristics or traits accepted by those skilled in the art as sufficient to distinguish one breeding line or line from another breeding line or line. There is no implication in either term that all plants of any given breeding line or line will be genetically identical at either the whole gene or molecular level or that any given plant will be homozygous at all loci. A breeding line or line is considered “true breeding” for a particular trait if, when the true-breeding line or breeding line is self-pollinated, all of the progeny contain the trait. In the present invention, the trait arises from a mutation in a PPO gene of the plant or seed.
In some embodiments, traditional plant breeding is employed whereby the uracilpyridine herbicides-tolerant trait is introduced in the progeny plant resulting therefrom. In one embodiment, the present invention provides a method for producing a uracilpyridine herbicides-tolerant progeny plant, the method comprising: crossing a parent plant with a uracilpyridine herbicides-tolerant plant to introduce the uracilpyridine herbicides-tolerance characteristics of the uracilpyridine herbicides-tolerant plant into the germplasm of the progeny plant, wherein the progeny plant has increased tolerance to the uracilpyridine herbicides relative to the parent plant. In other embodiments, the method further comprises the step of introgressing the uracilpyridine herbicides-tolerance characteristics through traditional plant breeding techniques to obtain a descendent plant having the uracilpyridine herbicides-tolerance characteristics.
Arabidopsis thaliana
The herbicide-resistant plants of the invention that comprise polynucleotides encoding mutated PPO polypeptides also find use in methods for increasing the herbicide-resistance of a plant through conventional plant breeding involving sexual reproduction. The methods comprise crossing a first plant that is a herbicide-resistant plant of the invention to a second plant that may or may not be resistant to the same herbicide or herbicides as the first plant or may be resistant to different herbicide or herbicides than the first plant. The second plant can be any plant that is capable of producing viable progeny plants (i.e., seeds) when crossed with the first plant. Typically, but not necessarily, the first and second plants are of the same species. The methods can optionally involve selecting for progeny plants that comprise the mutated PPO polypeptides of the first plant and the herbicide resistance characteristics of the second plant. The progeny plants produced by this method of the present invention have increased resistance to a herbicide when compared to either the first or second plant or both. When the first and second plants are resistant to different herbicides, the progeny plants will have the combined herbicide tolerance characteristics of the first and second plants. The methods of the invention can further involve one or more generations of backcrossing the progeny plants of the first cross to a plant of the same line or genotype as either the first or second plant. Alternatively, the progeny of the first cross or any subsequent cross can be crossed to a third plant that is of a different line or genotype than either the first or second plant. The present invention also provides plants, plant organs, plant tissues, plant cells, seeds, and non-human host cells that are transformed with the at least one polynucleotide molecule, expression cassette, or transformation vector of the invention. Such transformed plants, plant organs, plant tissues, plant cells, seeds, and non-human host cells have enhanced tolerance or resistance to at least one herbicide, at levels of the herbicide that kill or inhibit the growth of an untransformed plant, plant tissue, plant cell, or non-human host cell, respectively. Preferably, the transformed plants, plant tissues, plant cells, and seeds of the invention are and crop plants.
In other aspects, plants of the invention include those plants which, in addition to being tolerant to uracilpyridine herbicides, have been subjected to further genetic modifications by breeding, mutagenesis or genetic engineering, e.g. have been rendered tolerant to applications of specific other classes of herbicides, such as AHAS inhibitors; auxinic herbicides; bleaching herbicides such as hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors or phytoene desaturase (PDS) inhibitors; EPSPS inhibitors such as glyphosate; glutamine synthetase (GS) inhibitors such as glufosinate; lipid biosynthesis inhibitors such as acetyl CoA carboxylase (ACCase) inhibitors; or oxynil {i.e. bromoxynil or ioxynil) herbicides as a result of conventional methods of breeding or genetic engineering, Thus, uracilpyridine herbicides-tolerant plants of the invention can be made resistant to multiple classes of herbicides through multiple genetic modifications, such as resistance to both glyphosate and glufosinate or to both glyphosate and a herbicide from another class such as HPPD inhibitors, AHAS inhibitors, or ACCase inhibitors. These herbicide resistance technologies are, for example, described in Pest Management Science (at volume, year, page):61, 2005, 246; 61, 2005, 258; 61, 2005, 277; 61, 2005, 269; 61, 2005, 286; 64, 2008, 326; 64, 2008, 332; Weed Science 57, 2009, 108; Australian Journal of Agricultural Research 58, 2007, 708; Science 316, 2007, 1185; and references quoted therein. For example, uracilpyridine herbicides-tolerant plants of the invention, in some embodiments, may be tolerant to ACCase inhibitors, such as “dims” {e.g., cycloxydim, sethoxydim, clethodim, or tepraloxydim), “fops” {e.g., clodinafop, diclofop, fluazifop, haloxyfop, or quizalofop), and “dens” (such as pinoxaden); to auxinic herbicides, such as dicamba; to EPSPS inhibitors, such as glyphosate; to other PPO inhibitors; and to GS inhibitors, such as glufosinate.
In addition to these classes of inhibitors, uracilpyridine herbicides-tolerant plants of the invention may also be tolerant to herbicides having other modes of action, for example, chlorophyll/carotenoid pigment inhibitors, cell membrane disrupters, photosynthesis inhibitors, cell division inhibitors, root inhibitors, shoot inhibitors, and combinations thereof.
Such tolerance traits may be expressed, e.g.: as mutant or wildtype PPO proteins, as mutant AHASL proteins, mutant ACCase proteins, mutant EPSPS proteins, or mutant glutamine synthetase proteins; or as mutant native, inbred, or transgenic aryloxyalkanoate dioxygenase (AAD or DHT), haloarylnitrilase (BXN), 2,2-dichloropropionic acid dehalogenase (DEH), glyphosate-N-acetyltransferase (GAT), glyphosate decarboxylase (GDC), glyphosate oxidoreductase (GOX), glutathione-S-transferase (GST), phosphinothricin acetyltransferase (PAT or bar), or CYP450s proteins having an herbicide-degrading activity.
uracilpyridine herbicides-tolerant plants hereof can also be stacked with other traits including, but not limited to, pesticidal traits such as Bt Cry and other proteins having pesticidal activity toward coleopteran, lepidopteran, nematode, or other pests; nutrition or nutraceutical traits such as modified oil content or oil profile traits, high protein or high amino acid concentration traits, and other trait types known in the art.
Bacillus
Bacillus thuringiensis
Photorhabdus
Furthermore, in other embodiments, uracilpyridine herbicides-tolerant plants are also covered which are, by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such characteristics, rendered able to synthesize one or more insecticidal proteins, especially those known from the bacterial genus , particularly from , such as [delta]-endotoxins, e.g. CryIA(b), CryIA(c), CryIF, CryIF(a2), CryIIA(b), CryIIIA, CryIIIB(b1) or Cry9c; vegetative insecticidal proteins (VIP), e.g. VIP1, VIP2, VIP3 or VIP3A; insecticidal proteins of bacteria colonizing nematodes, e.g. spp. or Xenorhabdus spp.; toxins produced by animals, such as scorpion toxins, arachnid toxins, wasp toxins, or other insect-specific neurotoxins; toxins produced by fungi, such streptomycete toxins; plant lectins, such as pea or barley lectins; agglutinins; proteinase inhibitors, such as trypsin inhibitors, serine protease inhibitors, patatin, cystatin or papain inhibitors; ribosome-inactivating proteins (RIP), such as ricin, maize-RIP, abrin, luffin, saporin or bryodin; steroid metabolism enzymes, such as 3-hydroxy-steroid oxidase, ecdysteroid-IDP-glycosyl-transferase, cholesterol oxidases, ecdysone inhibitors or HMG-CoA-reductase; ion channel blockers, such as blockers of sodium or calcium channels; juvenile hormone esterase; diuretic hormone receptors (helicokinin receptors); stilben synthase, bibenzyl synthase, chitinases or glucanases. In the context of the present invention these insecticidal proteins or toxins are to be understood expressly also as pre-toxins, hybrid proteins, truncated or otherwise modified proteins. Hybrid proteins are characterized by a new combination of protein domains, (see, e.g. WO 02/015701). Further examples of such toxins or genetically modified plants capable of synthesizing such toxins are disclosed, e.g., in EP-A 374 753, WO 93/007278, WO 95/34656, EP-A 427 529, EP-A 451 878, WO 03/18810 und WO 03/52073. The methods for producing such genetically modified plants are generally known to the person skilled in the art and are described, e.g. in the publications mentioned above. These insecticidal proteins contained in the genetically modified plants impart to the plants producing these proteins tolerance to harmful pests from all taxonomic groups of arthropods, especially to beetles (Coeloptera), two-winged insects (Diptera), and moths (Lepidoptera) and to nematodes (Nematoda).
Acanthoscelides obtectus
Agriotes lineatus, Agriotes obscurus, Agriotes
solstitialis
Anthonomus
Atomaria linearis
Anthrenus
Attagenus
Carpophilus hemipterus
Ceutorhynchus assimilis
Ceutorhynchus picitarsis
Conoderus vespertinus
Conoderus falli
Cosmopolites sordidus
Costelytra zealandica
Cotinis nitida
Cylindrocopturus adspersus
Dermestes lardarius
Diabrotica virgifera, Diabrotica virgifera virgifera
Diabrotica barberi
Epilachna varivestis
Hylotropes bajulus
Hypera postica
Gibbium psylloides
Lasioderma serricorne
Leptinotarsa decemlineata
Lyctus
Meligethes aeneus
Melolontha melolontha
Mezium americanum
Oryzaephilus surinamensis
Oryzaephilus Mercator
Otiorhynchus sulcatus
Phaedon cochleariae
Phyllotreta cruciferae
Phyllotreta striolata
Psylliodes chrysocephala; Ptinus
Rhizopertha dominica
Sitona lineatus
Sitophilus oryzae
Sitophilus granaries
Smicronyx fulvus
Stegobium paniceum
Tenebrio molitor
Tribolium castaneum
Tribolium confusum
Trogoderma
Zygogramma exclamationis; Dermaptera
Forficula auricularia
Labidura riparia; Dictyoptera
Blatta orientalis
Oxidus gracilis
Pegomyia betae
Oscinella frit
Dacus
Drosophila
Isoptera
Lygus lineolaris
Aphis fabae
Aphis gossypii
Aphis pomi
Aleurocanthus spiniferus
Bemesia tabaci
Brevicoryne brassicae
psylla Cacopsylla pyricola
Cryptomyzus ribis
phylloxera Daktulosphaira vitifoliae
psylla Diaphorina citri
Empoasca fabae
Empoasca
Empoasca vitis
Eriosoma lanigerum
Eulecanium corni
Hyalopterus arundinis
Laodelphax striatellus
Macrosiphum euphorbiae
Myzus persicae
Nephotettix cinticeps
Nilaparvata lugens
Phorodon humuli
Rhopalosiphum padi
Sitobion avenae
Adoxophyes orana
tortrix
Archips podana
tortrix
Bucculatrix
Bucculatrix thurberiella
Bupalus piniarius
Carpocapsa pomonella
Chilo suppressalis
Choristoneura fumiferana
Cochylis hospes
Diatraea grandiosella
Eupoecilia ambiguella
Helicoverpa armigera
Helicoverpa zea
Heliothis
Homeosoma electellum
Homona magnanima
tortrix
Lithocolletis blancardella
Lymantria dispar
Malacosoma neustria
Mamestra brassicae
Mamestra configurata
Operophtera brumata
Ostrinia nubilalis
Panolis flammea
Phyllocnistis citrella
Pieris brassicae
Rachiplusia ni
Spodoptera exigua
Spodoptera littoralis
Sylepta derogata
Trichoplusia ni
Orthoptera
Acheta domesticus
Anacridium
Locusta migratoria
Melanoplus bivittatus
Melanoplus
Melanoplus femurrubrum
Melanoplus sanguinipes
Neocurtilla hexadectyla
Nomadacris septemfasciata
Scapteriscus abbreviatus
Scapteriscus borellii
Scapteriscus vicinus
Schistocerca gregaria; Symphyla
Scutigerella immaculata; Thysanoptera
thrips Frankliniella fusca
thrips Frankliniella intonsa
thrips Frankliniella
thrips Frankliniella schultzei
thrips Hercinothrips femoralis
thrips Neohydatothrips variabilis
thrips Pezothrips kellyanus
thrips Scirtothrips perseae
thrips Thrips palmi
thrips Thrips tabaci
In some embodiments, expression of one or more protein toxins (e.g., insecticidal proteins) in the uracilpyridine herbicides-tolerant plants is effective for controlling organisms that include, for example, members of the classes and orders: Coleoptera such as the American bean weevil ; the leaf beetle Agelastica alni; click beetles (bicolor); the grain beetle Ahasverus advena; the summer schafer Amphimallon ; the furniture beetle Anobium punctatum; spp. (weevils); the Pygmy mangold beetle ; carpet beetles (spp., spp.); the cowpea weevil Callosobruchus maculates; the fried fruit beetle ; the cabbage seedpod weevil ; the rape winter stem weevil ; the wireworms and ; the banana weevil ; the New Zealand grass grub ; the June beetle ; the sunflower stem weevil ; the larder beetle ; the corn rootworms , and ; the Mexican bean beetle ; the old house borer ; the lucerne weevil ; the shiny spider beetle ; the cigarette beetle ; the Colorado potato beetle ; Lyctus beetles {spp., the pollen beetle ; the common cockshafer ; the American spider beetle ; the golden spider beetle Niptus hololeuc s; the grain beetles and ; the black vine weevil ; the mustard beetle , the crucifer flea beetle ; the striped flea beetle ; the cabbage steam flea beetle spp. (spider beetles); the lesser grain borer ; the pea and been weevil ; the rice and granary beetles and ; the red sunflower seed weevil ; the drugstore beetle ; the yellow mealworm beetle , the flour beetles and ; warehouse and cabinet beetles {spp.); the sunflower beetle (earwigs) such as the European earwig and the striped earwig such as the oriental cockroach ; the greenhouse millipede ; the beet fly ; the frit fly ; fruitflies (spp., spp.); (termites) including species from the familes Hodotermitidae, Kalotermitidae, Mastotermitidae, Rhinotermitidae, Serritermitidae, Termitidae, Termopsidae; the tarnished plant bug ; the black bean aphid ; the cotton or melon aphid ; the green apple aphid ; the citrus spiny whitefly ; the sweet potato whitefly ; the cabbage aphid ; the pear ; the currant aphid ; the grape ; the citrus ; the potato leafhopper ; the bean leafhopper Solana; the vine leafhopper ; the woolly aphid ; the European fruit scale ; the mealy plum aphid ; the small brown planthopper ; the potato aphid ; the green peach aphid ; the green rice leafhopper ; the brown planthopper ; the hop aphid ; the bird-cherry aphid ; the grain aphid ; Lepidoptera such as (summer fruit moth); (fruit tree moth); pyrivorella (pear leafminer); (cotton leaf perforator); (pine looper); (codling moth); (striped rice borer); (eastern spruce budworm); (banded sunflower moth); (southwestern corn borer); (European grape berry moth); (cotton bollworm); (cotton bollworm); vires cens (tobacco budworm), (sunflower moth); (oriental tea tree moth); (spotted tentiform leafminer); (gypsy moth); (tent caterpillar); (cabbage armyworm); (Bertha armyworm); (winter moth); (European corn borer), (pine beauty moth), (citrus leafminer); (cabbage white butterfly); (soybean looper); (beet armyworm); (cotton leafworm); (cotton leaf roller); (cabbage looper); such as the common cricket , tree locusts (spp.), the migratory locust , the twostriped grasshopper , the differential grasshopper differ entialis, the redlegged grasshopper , the migratory grasshopper , the northern mole cricket , the red locust , the shortwinged mole cricket , the southern mole cricket , the tawny mole cricket , and the desert locust such as the garden symphylan such as the tobacco , the flower , the western flower occidentalism the cotton bud , the banded greenhouse , the soybean , Kelly's citrus , the avocado , the melon , and the onion ; and the like, and combinations comprising one or more of the foregoing organisms.
Phyllotreta
Phyllotreta cruciferae
Phyllotreta triolata
Lygus
In some embodiments, expression of one or more protein toxins (e.g., insecticidal proteins) in the uracilpyridine herbicides-tolerant plants is effective for controlling flea beetles, i.e. members of the flea beetle tribe of family Chrysomelidae, preferably against spp., such as and/or . In other embodiments, expression of one or more protein toxins {e.g., insecticidal proteins) in the uracilpyridine herbicides-tolerant plants is effective for controlling cabbage seedpod weevil, the Bertha armyworm, bugs, or the diamondback moth.
Furthermore, in one embodiment, uracilpyridine herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, rendered able to synthesize one or more proteins to increase the resistance or tolerance of those plants to bacterial, viral or fungal pathogens. The methods for producing such genetically modified plants are generally known to the person skilled in the art.
Furthermore, in another embodiment, uracilpyridine herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, rendered able to synthesize one or more proteins to increase the productivity (e.g. oil content), tolerance to drought, salinity or other growth-limiting environmental factors or tolerance to pests and fungal, bacterial or viral pathogens of those plants.
Furthermore, in other embodiments, uracilpyridine herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, altered to contain a modified amount of one or more substances or new substances, for example, to improve human or animal nutrition, e.g. oil crops that produce health-promoting long-chain omega-3 fatty acids or unsaturated omega-9 fatty acids (e.g. Nexera® rape, Dow Agro Sciences, Canada).
Furthermore, in some embodiments, uracilpyridine herbicides-tolerant plants are also covered which are, e.g. by the use of recombinant DNA techniques and/or by breeding and/or otherwise selected for such traits, altered to contain increased amounts of vitamins and/or minerals, and/or improved profiles of nutraceutical compounds.
In one embodiment, uracilpyridine herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: glucosinolates (e.g., glucoraphanin (4-methylsulfinylbutyl-glucosinolate), sulforaphane, 3-indolylmethyl-glucosinolate(glucobrassicin), I-methoxy-3-indolylmethyl-glucosinolate (neoglucobrassicin)); phenolics (e.g., flavonoids (e.g., quercetin, kaempferol), hydroxycinnamoyl derivatives (e.g., 1,2,2′-trisinapoylgentiobiose, 1,2-diferuloylgentiobiose, I,2′-disinapoyl-2-feruloylgentiobiose, 3-0-caffeoyl-quinic (neochlorogenic acid)); and vitamins and minerals (e.g., vitamin C, vitamin E, carotene, folic acid, niacin, riboflavin, thiamine, calcium, iron, magnesium, potassium, selenium, and zinc).
In another embodiment, uracilpyridine herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: progoitrin; isothiocyanates; indoles (products of glucosinolate hydrolysis); glutathione; carotenoids such as beta-carotene, lycopene, and the xanthophyll carotenoids such as lutein and zeaxanthin; phenolics comprising the flavonoids such as the flavonols (e.g. quercetin, rutin), the flavans/tannins (such as the procyanidins comprising coumarin, proanthocyanidins, catechins, and anthocyanins); flavones; phytoestrogens such as coumestans, lignans, resveratrol, isoflavones e.g. genistein, daidzein, and glycitein; resorcyclic acid lactones; organosulphur compounds; phytosterols; terpenoids such as carnosol, rosmarinic acid, glycyrrhizin and saponins; chlorophyll; chlorphyllin, sugars, anthocyanins, and vanilla. In other embodiments, uracilpyridine herbicides-tolerant plants of the present invention, relative to a wild-type plant, comprise an increased amount of, or an improved profile of, a compound selected from the group consisting of: vincristine, vinblastine, taxanes (e.g., taxol (paclitaxel), baccatin III, 10-desacetylbaccatin III, 10-desacetyl taxol, xylosyl taxol, 7-epitaxol, 7-epibaccatin III, 10-desacetylcephalomannine, 7-epicephalomannine, taxotere, cephalomannine, xylosyl cephalomannine, taxagifine, 8-benxoyloxy taxagifine, 9-acetyloxy taxusin, 9-hydroxy taxusin, taiwanxam, taxane Ia, taxane Ib, taxane Ic, taxane Id, GMP paclitaxel, 9-dihydro 13-acetylbaccatin III, 10-desacetyl-7-epitaxol, tetrahydrocannabinol (THC), cannabidiol (CBD), genistein, diadzein, codeine, morphine, quinine, shikonin, ajmalacine, serpentine, and the like.
It is to be understood that the plant of the present invention can comprise a wild type PPO nucleic acid in addition to a mutated PPO nucleic acid. It is contemplated that the uracilpyridine herbicide tolerant lines may contain a mutation in only one of multiple PPO isoenzymes. Therefore, the present invention includes a plant comprising one or more mutated PPO nucleic acids in addition to one or more wild type PPO nucleic acids.
In another embodiment, the invention refers to a seed produced by a transgenic plant comprising a plant cell of the present invention, wherein the seed is true breeding for an increased resistance to a uracilpyridine herbicide as compared to a wild type variety of the seed.
In another embodiment, the invention refers to a method of producing a transgenic plant cell with an increased resistance to a uracilpyridine herbicide as compared to a wild type variety of the plant cell comprising, transforming the plant cell with an expression cassette comprising a mutated PPO nucleic acid.
In another embodiment, the invention refers to a method of producing a transgenic plant comprising, (a) transforming a plant cell with an expression cassette comprising a mutated PPO nucleic acid, and (b) generating a plant with an increased resistance to uracilpyridine herbicide from the plant cell.
Consequently, mutated PPO nucleic acids of the invention are provided in expression cassettes for expression in the plant of interest. The cassette will include regulatory sequences operably linked to a mutated PPO nucleic acid sequence of the invention. The term “regulatory element” as used herein refers to a polynucleotide that is capable of regulating the transcription of an operably linked polynucleotide. It includes, but not limited to, promoters, enhancers, introns, 5′ UTRs, and 3′ UTRs. By “operably linked” is intended a functional linkage between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences being linked are contiguous and, where necessary to join two protein coding regions, contiguous and in the same reading frame. The cassette may additionally contain at least one additional gene to be cotransformed into the organism. Alternatively, the additional gene(s) can be provided on multiple expression cassettes.
In a preferred embodiment, the expression cassette comprises the sequence of SEQ ID NO:240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 264.
Such an expression cassette is provided with a plurality of restriction sites for insertion of the mutated PPO nucleic acid sequence to be under the transcriptional regulation of the regulatory regions. The expression cassette may additionally contain selectable marker genes.
The expression cassette of the present invention will include in the 5′-3′ direction of transcription, a transcriptional and translational initiation region (i.e., a promoter), a mutated PPO encoding nucleic acid sequence of the invention, and a transcriptional and translational termination region (i.e., termination region) functional in plants. The promoter may be native or analogous, or foreign or heterologous, to the plant host and/or to the mutated PPO nucleic acid sequence of the invention. Additionally, the promoter may be the natural sequence or alternatively a synthetic sequence. Where the promoter is “foreign” or “heterologous” to the plant host, it is intended that the promoter is not found in the native plant into which the promoter is introduced. Where the promoter is “foreign” or “heterologous” to the mutated PPO nucleic acid sequence of the invention, it is intended that the promoter is not the native or naturally occurring promoter for the operably linked mutated PPO nucleic acid sequence of the invention. As used herein, a chimeric gene comprises a coding sequence operably linked to a transcription initiation region that is heterologous to the coding sequence.
While it may be preferable to express the mutated PPO nucleic acids of the invention using heterologous promoters, the native promoter sequences may be used. Such constructs would change expression levels of the mutated PPO protein in the plant or plant cell. Thus, the phenotype of the plant or plant cell is altered.
A. tumefaciens
The termination region may be native with the transcriptional initiation region, may be native with the operably linked mutated PPO sequence of interest, may be native with the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the mutated PPO nucleic acid sequence of interest, the plant host, or any combination thereof). Convenient termination regions are available from the Ti-plasmid of , such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262: 141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991) Genes Dev. 5: 141-149; Mogen et al. (1990) Plant Cell 2: 1261-1272; Munroe et al. (1990) Gene 91: 151-158; Ballas t al. (1989) Nucleic Acids Res. 17:7891-7903; and Joshi et al. (1987) Nucleic Acid Res. 15:9627-9639. Where appropriate, the gene(s) may be optimized for increased expression in the transformed plant. That is, the genes can be synthesized using plant-preferred codons for improved expression. See, for example, Campbell and Gowri (1990) Plant Physiol. 92: 1-11 for a discussion of host-preferred codon usage. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Pat. Nos. 5,380,831, and 5,436,391, and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference.
Additional sequence modifications are known to enhance gene expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon-intron splice site signals, transposon-like repeats, and other such well-characterized sequences that may be deleterious to gene expression. The G-C content of the sequence may be adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell. When possible, the sequence is modified to avoid predicted hairpin secondary mRNA structures. Nucleotide sequences for enhancing gene expression can also be used in the plant expression vectors. These include the introns of the maize Adhl, intronl gene (Callis et al. Genes and Development 1: 1183-1200, 1987), and leader sequences, (W-sequence) from the Tobacco Mosaic virus (TMV), Maize Chlorotic Mottle Virus and Alfalfa Mosaic Virus (Gallie et al. Nucleic Acid Res. 15:8693-8711, 1987 and Skuzeski et al. Plant Mol. Biol. 15:65-79, 1990). The first intron from the shrunken-1 locus of maize, has been shown to increase expression of genes in chimeric gene constructs. U.S. Pat. Nos. 5,424,412 and 5,593,874 disclose the use of specific introns in gene expression constructs, and Gallie et al. (Plant Physiol. 106:929-939, 1994) also have shown that introns are useful for regulating gene expression on a tissue specific basis. To further enhance or to optimize mutated PPO gene expression, the plant expression vectors of the invention may also contain DNA sequences containing matrix attachment regions (MARs). Plant cells transformed with such modified expression systems, then, may exhibit overexpression or constitutive expression of a nucleotide sequence of the invention.
The expression cassettes of the present invention may additionally contain 5′ leader sequences in the expression cassette construct. Such leader sequences can act to enhance translation. Translation leaders are known in the art and include: picornavirus leaders, for example, EMCV leader (Encephalomyocarditis 5′ noncoding region) (Elroy-Stein et al. (1989) Proc. Natl. Acad. ScL USA 86:6126-6130); potyvirus leaders, for example, TEV leader (Tobacco Etch Virus) (Gallie et al. (1995) Gene 165(2):233-238), MDMV leader (Maize Dwarf Mosaic Virus) (Virology 154:9-20), and human immunoglobulin heavy-chain binding protein (BiP) (Macejak et al. (1991) Nature 353:90-94); untranslated leader from the coat protein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling et al. (1987) Nature 325:622-625); tobacco mosaic virus leader (TMV) (Gallie et al. (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York), pp. 237-256); and maize chlorotic mottle virus leader (MCMV) (Lommel et al. (1991) Virology 81:382-385). See also, Della-Cioppa et al. (1987) Plant Physiol. 84:965-968. Other methods known to enhance translation can also be utilized, for example, introns, and the like.
In preparing the expression cassette, the various DNA fragments may be manipulated, so as to provide for the DNA sequences in the proper orientation and, as appropriate, in the proper reading frame. Toward this end, adapters or linkers may be employed to join the DNA fragments or other manipulations may be involved to provide for convenient restriction sites, removal of superfluous DNA, removal of restriction sites, or the like. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, resubstitutions, e.g., transitions and trans versions, may be involved.
A number of promoters can be used in the practice of the invention. The promoters can be selected based on the desired outcome. The nucleic acids can be combined with constitutive, tissue-preferred, or other promoters for expression in plants. Such constitutive promoters include, for example, the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO 99/43838 and U.S. Pat. No. 6,072,050; the core CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2: 163-171); ubiquitin (Christensen et al. (1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) Plant Mol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet. 81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730); ALS promoter (U.S. Pat. No. 5,659,026), and the like. Other constitutive promoters include, for example, U.S. Pat. Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611.
Tissue-preferred promoters can be utilized to target enhanced mutated PPO expression within a particular plant tissue. Such tissue-preferred promoters include, but are not limited to, leaf-preferred promoters, root-preferred promoters, seed-preferred promoters, and stem-preferred promoters. Tissue-preferred promoters include Yamamoto et al. (1997) Plant J. 12(2):255-265; Kawamata et al. (1997) Plant Cell Physiol. 38(7):792-803; Hansen et al. (1997) Mol. Gen Genet. 254(3):337-343; Russell et al. (1997) Transgenic Res. 6(2): 157-168; Rinehart et al. (1996) Plant Physiol. 112(3): 1331-1341; Van Camp et al. (1996) Plant Physiol. 112(2):525-535; Canevascini et al. (1996) Plant Physiol. 112(2):513-524; Yamamoto et al. (1994) Plant Cell Physiol. 35(5):773-778; Lam (1994) Results Probl. Cell Differ. 20: 181-196; Orozco et al. (1993) Plant Mol Biol. 23(6): 1129-1138; Matsuoka e/[alpha]/. (1993) Proc Natl. Acad. Sci. USA 90(20):9586-9590; and Guevara-Garcia et al. (1993) Plant J. 4(3):495-505. Such promoters can be modified, if necessary, for weak expression. In one embodiment, the nucleic acids of interest are targeted to the chloroplast for expression. In this manner, where the nucleic acid of interest is not directly inserted into the chloroplast, the expression cassette will additionally contain a chloroplast-targeting sequence comprising a nucleotide sequence that encodes a chloroplast transit peptide to direct the gene product of interest to the chloroplasts. Such transit peptides are known in the art. With respect to chloroplast-targeting sequences, “operably linked” means that the nucleic acid sequence encoding a transit peptide (i.e., the chloroplast-targeting sequence) is linked to the mutated PPO nucleic acid of the invention such that the two sequences are contiguous and in the same reading frame. See, for example, Von Heijne et al. (1991) Plant Mol. Biol. Rep. 9: 104-126; Clark et al. (1989) J. Biol. Chem. 264:17544-17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem. Biophys. Res. Commun. 196:1414-1421; and Shah et al. (1986) Science 233:478-481. While the mutated PPO proteins of the invention include a native chloroplast transit peptide, any chloroplast transit peptide known in the art can be fused to the amino acid sequence of a mature mutated PPO protein of the invention by operably linking a choloroplast-targeting sequence to the 5′-end of a nucleotide sequence encoding a mature mutated PPO protein of the invention. Chloroplast targeting sequences are known in the art and include the chloroplast small subunit of ribulose-1,5-bisphosphate carboxylase (Rubisco) (de Castro Silva Filho et al. (1996) Plant Mol. Biol. 30:769-780; Schnell et al. (1991) J. Biol. Chem. 266(5):3335-3342); 5-(enolpyruvyl)shikimate-3-phosphate synthase (EPSPS) (Archer et al. (1990) J. Bioenerg. Biomemb. 22(6):789-810); tryptophan synthase (Zhao et al. (1995) J. Biol. Chem. 270(11):6081-6087); plastocyanin (Lawrence et al. (1997) J. Biol. Chem. 272(33):20357-20363); chorismate synthase (Schmidt et al. (1993) J. Biol. Chem. 268(36):27447-27457); and the light harvesting chlorophyll a/b binding protein (LHBP) (Lamppa et al. (1988) J. Biol. Chem. 263: 14996-14999). See also Von Heijne et al. (1991) Plant Mol. Biol. Rep. 9: 104-126; Clark et al. (1989) J. Biol. Chem. 264:17544-17550; Della-Cioppa et al. (1987) Plant Physiol. 84:965-968; Romer et al. (1993) Biochem. Biophys. Res. Commun. 196: 1414-1421; and Shah et al. (1986) Science 233:478-481.
In a preferred embodiment, the transit peptide comprises the amino acid sequence of SEQ ID NO: 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226.
Amaranthus
Preferred expression cassettes comprise a transit peptide from PPO-2 fused to PPO or PPO like polypeptides, such as chimeric expression cassettes having the SEQ ID NO:240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262,or 264
Methods for transformation of chloroplasts are known in the art. See, for example, Svab et al. (1990) Proc. Natl. Acad. ScL USA 87:8526-8530; Svab and Maliga (1993) Proc. Natl. Acad. Sci. USA 90:913-917; Svab and Maliga (1993) EMBO J. 12:601-606. The method relies on particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination. Additionally, plastid transformation can be accomplished by transactivation of a silent plastid-borne transgene by tissue-preferred expression of a nuclear-encoded and plastid-directed RNA polymerase. Such a system has been reported in McBride et al. (1994) Proc. Natl. Acad. Sci. USA 91:7301-7305. The nucleic acids of interest to be targeted to the chloroplast may be optimized for expression in the chloroplast to account for differences in codon usage between the plant nucleus and this organelle. In this manner, the nucleic acids of interest may be synthesized using chloroplast-preferred codons. See, for example, U.S. Pat. No. 5,380,831, herein incorporated by reference.
In a preferred embodiment, the mutated PPO nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide encoding a polypeptide as shown in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, or 637,or a variant or derivative thereof; b) a polynucleotide comprising at least 60 consecutive nucleotides of any of a); and c) a polynucleotide complementary to the polynucleotide of any of a) through b)
Preferably, the expression cassette of the present invention further comprises a transcription initiation regulatory region and a translation initiation regulatory region that are functional in the plant.
While the polynucleotides of the invention find use as selectable marker genes for plant transformation, the expression cassettes of the invention can include another selectable marker gene for the selection of transformed cells. Selectable marker genes, including those of the present invention, are utilized for the selection of transformed cells or tissues. Marker genes include, but are not limited to, genes encoding antibiotic resistance, such as those encoding neomycin phosphotransferase II (NEO) and hygromycin phosphotransferase (HPT), as well as genes conferring resistance to herbicidal compounds, such as glufosinate ammonium, bromoxynil, imidazolinones, and 2,4-dichlorophenoxyacetate (2,4-D). See generally, Yarranton (1992) Curr. Opin. Biotech. 3: 506-511; Christophers on et al (1992) Proc. Natl. Acad. ScL USA 89:6314-6318; Yao et al. (1992) Cell 71:63-72; Reznikoff (1992) Mol Microbiol 6:2419-2422; Barkley et al (1980) in The Operon, pp. 177-220; Hu et al (1987) Cell 48:555-566; Brown et al (1987) Cell 49:603-612; Figge et al (1988) Cell 52:713-722; Deuschle et al (1989) Proc. Natl Acad. AcL USA 86:5400-5404; Fuerst et al (1989) Proc. Natl Acad. ScL USA 86:2549-2553; Deuschle et al (1990) Science 248:480-483; Gossen (1993) Ph.D. Thesis, University of Heidelberg; Reines et al (1993) Proc. Natl Acad. ScL USA 90: 1917-1921; Labow et al (1990) Mol Cell Biol 10:3343-3356; Zambretti et al (1992) Proc. Natl Acad. ScL USA 89:3952-3956; Bairn et al (1991) Proc. Natl Acad. ScL USA 88:5072-5076; Wyborski et al (1991) Nucleic Acids Res. 19:4647-4653; Hillenand-Wissman (1989) Topics Mol Struc. Biol 10: 143-162; Degenkolb et al (1991) Antimicrob. Agents Chemother. 35: 1591-1595; Kleinschnidt et al (1988) Biochemistry 27: 1094-1104; Bonin (1993) Ph.D. Thesis, University of Heidelberg; Gossen et al (1992) Proc. Natl Acad. ScL USA 89:5547-5551; Oliva et al (1992) Antimicrob. Agents Chemother. 36:913-919; Hlavka et al (1985) Handbook of Experimental Pharmacology, Vol. 78 (Springer-Verlag, Berlin); Gill et al (1988) Nature 334:721-724. Such disclosures are herein incorporated by reference. The above list of selectable marker genes is not meant to be limiting. Any selectable marker gene can be used in the present invention.
The invention further provides an isolated recombinant expression vector comprising the expression cassette containing a mutated PPO nucleic acid as described above, wherein expression of the vector in a host cell results in increased tolerance to a uracilpyridine herbicide as compared to a wild type variety of the host cell. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “expression vectors.” In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, “plasmid” and “vector” can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), which serve equivalent functions.
The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operably linked to the nucleic acid sequence to be expressed. Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of the nucleotide sequence only in certain host cells or under certain conditions. It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce polypeptides or peptides, including fusion polypeptides or peptides, encoded by nucleic acids as described herein (e.g., mutated PPO polypeptides, fusion polypeptides, etc.).
In a preferred embodiment of the present invention, the mutated PPO polypeptides are expressed in plants and plants cells such as unicellular plant cells (such as algae) (See Falciatore et al., 1999, Marine Biotechnology 1(3):239-251 and references therein) and plant cells from higher plants (e.g., the spermatophytes, such as crop plants). A mutated PPO polynucleotide may be “introduced” into a plant cell by any means, including transfection, transformation or transduction, electroporation, particle bombardment, agroinfection, biolistics, and the like.
Agrobacterium
manihot
tagetes
Vicia
Suitable methods for transforming or transfecting host cells including plant cells can be found in Sambrook et al. (Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989) and other laboratory manuals such as Methods in Molecular Biology, 1995, Vol. 44, protocols, ed: Gartland and Davey, Humana Press, Totowa, N.J. As increased tolerance to uracilpyridine herbicides is a general trait wished to be inherited into a wide variety of plants like maize, wheat, rye, oat, triticale, rice, barley, soybean, peanut, cotton, rapeseed and canola, , pepper, sunflower and , solanaceous plants like potato, tobacco, eggplant, and tomato, species, pea, alfalfa, bushy plants (coffee, cacao, tea), Salix species, trees (oil palm, coconut), perennial grasses, and forage crops, these crop plants are also preferred target plants for a genetic engineering as one further embodiment of the present invention. In a preferred embodiment, the plant is a crop plant. Forage crops include, but are not limited to, Wheatgrass, Canarygrass, Bromegrass, Wildrye Grass, Bluegrass, Orchardgrass, Alfalfa, Salfoin, Birdsfoot Trefoil, Alsike Clover, Red Clover, and Sweet Clover.
Agrobacterium
Agrobacterium
Agrobacterium tumefaciens
Agrobacterium
Agrobacterium
Agrobacterium
In one embodiment of the present invention, transfection of a mutated PPO polynucleotide into a plant is achieved by mediated gene transfer. One transformation method known to those of skill in the art is the dipping of a flowering plant into an Agrobacteria solution, wherein the Agrobacteria contains the mutated PPO nucleic acid, followed by breeding of the transformed gametes. mediated plant transformation can be performed using for example the GV3101(pMP90) (Koncz and Schell, 1986, Mol. Gen. Genet. 204:383-396) or LBA4404 (Clontech) strain. Transformation can be performed by standard transformation and regeneration techniques (Deblaere et al., 1994, Nucl. Acids. Res. 13:4777-4788; Gelvin, Stanton B. and Schilperoort, Robert A, Plant Molecular Biology Manual, 2nd Ed.—Dordrecht: Kluwer Academic Publ., 1995.— in Sect., Ringbuc Zentrale Signatur: BT11-P ISBN 0-7923-2731-4; Glick, Bernard R. and Thompson, John E., Methods in Plant Molecular Biology and Biotechnology, Boca Raton: CRC Press, 1993 360 S., ISBN 0-8493-5164-2). For example, rapeseed can be transformed via cotyledon or hypocotyl transformation (Moloney et al., 1989, Plant Cell Report 8:238-242; De Block et al., 1989, Plant Physiol. 91:694-701). Use of antibiotics for and plant selection depends on the binary vector and the strain used for transformation. Rapeseed selection is normally performed using kanamycin as selectable plant marker. mediated gene transfer to flax can be performed using, for example, a technique described by Mlynarova et al., 1994, Plant Cell Report 13:282-285. Additionally, transformation of soybean can be performed using for example a technique described in European Patent No. 0424 047, U.S. Pat. No. 5,322,783, European Patent No. 0397 687, U.S. Pat. No. 5,376,543, or U.S. Pat. No. 5,169,770. Transformation of maize can be achieved by particle bombardment, polyethylene glycol mediated DNA uptake, or via the silicon carbide fiber technique. (See, for example, Freeling and Walbot “The maize handbook” Springer Verlag: New York (1993) ISBN 3-540-97826-7). A specific example of maize transformation is found in U.S. Pat. No. 5,990,387, and a specific example of wheat transformation can be found in PCT Application No. WO 93/07256.
Triticum
According to the present invention, the introduced mutated PPO polynucleotide may be maintained in the plant cell stably if it is incorporated into a non-chromosomal autonomous replicon or integrated into the plant chromosomes. Alternatively, the introduced mutated PPO polynucleotide may be present on an extra-chromosomal non-replicating vector and be transiently expressed or transiently active. In one embodiment, a homologous recombinant microorganism can be created wherein the mutated PPO polynucleotide is integrated into a chromosome, a vector is prepared which contains at least a portion of an PPO gene into which a deletion, addition, or substitution has been introduced to thereby alter, e.g., functionally disrupt, the endogenous PPO gene and to create a mutated PPO gene. To create a point mutation via homologous recombination, DNA-RNA hybrids can be used in a technique known as chimeraplasty (Cole-Strauss et al., 1999, Nucleic Acids Research 27(5):1323-1330 and Kmiec, 1999, Gene therapy American Scientist 87(3):240-247). Other homologous recombination procedures in species are also well known in the art and are contemplated for use herein.
Physcomitrella patens
In the homologous recombination vector, the mutated PPO gene can be flanked at its 5′ and 3′ ends by an additional nucleic acid molecule of the PPO gene to allow for homologous recombination to occur between the exogenous mutated PPO gene carried by the vector and an endogenous PPO gene, in a microorganism or plant. The additional flanking PPO nucleic acid molecule is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several hundreds of base pairs up to kilobases of flanking DNA (both at the 5′ and 3′ ends) are included in the vector (see e.g., Thomas, K. R., and Capecchi, M. R., 1987, Cell 51:503 for a description of homologous recombination vectors or Strepp et al., 1998, PNAS, 95(8):4368-4373 for cDNA based recombination in ). However, since the mutated PPO gene normally differs from the PPO gene at very few amino acids, a flanking sequence is not always necessary. The homologous recombination vector is introduced into a microorganism or plant cell (e.g., via polyethylene glycol mediated DNA), and cells in which the introduced mutated PPO gene has homologously recombined with the endogenous PPO gene are selected using art-known techniques.
In another embodiment, recombinant microorganisms can be produced that contain selected systems that allow for regulated expression of the introduced gene. For example, inclusion of a mutated PPO gene on a vector placing it under control of the lac operon permits expression of the mutated PPO gene only in the presence of IPTG. Such regulatory systems are well known in the art.
C. glutamicum
C. glutamicum
Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but they also apply to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein. A host cell can be any prokaryotic or eukaryotic cell. For example, a mutated PPO polynucleotide can be expressed in bacterial cells such as , insect cells, fungal cells, or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells), algae, ciliates, plant cells, fungi or other microorganisms like . Other suitable host cells are known to those skilled in the art.
A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) a mutated PPO polynucleotide. Accordingly, the invention further provides methods for producing mutated PPO polypeptides using the host cells of the invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding a mutated PPO polypeptide has been introduced, or into which genome has been introduced a gene encoding a wild-type or mutated PPO polypeptide) in a suitable medium until mutated PPO polypeptide is produced. In another embodiment, the method further comprises isolating mutated PPO polypeptides from the medium or the host cell. Another aspect of the invention pertains to isolated mutated PPO polypeptides, and biologically active portions thereof. An “isolated” or “purified” polypeptide or biologically active portion thereof is free of some of the cellular material when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. The language “substantially free of cellular material” includes preparations of mutated PPO polypeptide in which the polypeptide is separated from some of the cellular components of the cells in which it is naturally or recombinantly produced. In one embodiment, the language “substantially free of cellular material” includes preparations of a mutated PPO polypeptide having less than about 30% (by dry weight) of non-mutated PPO material (also referred to herein as a “contaminating polypeptide”), more preferably less than about 20% of non-mutated PPO material, still more preferably less than about 10% of non-mutated PPO material, and most preferably less than about 5% non-mutated PPO material.
C. glutamicum
When the mutated PPO polypeptide, or biologically active portion thereof, is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the volume of the polypeptide preparation. The language “substantially free of chemical precursors or other chemicals” includes preparations of mutated PPO polypeptide in which the polypeptide is separated from chemical precursors or other chemicals that are involved in the synthesis of the polypeptide. In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of a mutated PPO polypeptide having less than about 30% (by dry weight) of chemical precursors or non-mutated PPO chemicals, more preferably less than about 20% chemical precursors or non-mutated PPO chemicals, still more preferably less than about 10% chemical precursors or non-mutated PPO chemicals, and most preferably less than about 5% chemical precursors or non-mutated PPO chemicals. In preferred embodiments, isolated polypeptides, or biologically active portions thereof, lack contaminating polypeptides from the same organism from which the mutated PPO polypeptide is derived. Typically, such polypeptides are produced by recombinant expression of, for example, a mutated PPO polypeptide in plants other than, or in microorganisms such as , ciliates, algae, or fungi.
In other aspects, a method for treating a plant of the present invention is provided.
In some embodiments, the method comprises contacting the plant with an agronomically acceptable composition.
In another aspect, the present invention provides a method for preparing a descendent seed. The method comprises planting a seed of or capable of producing a plant of the present invention. In one embodiment, the method further comprises growing a descendent plant from the seed; and harvesting a descendant seed from the descendent plant. In other embodiments, the method further comprises applying a uracilpyridine herbicides herbicidal composition to the descendent plant.
In another embodiment, the invention refers to harvestable parts of the transgenic plant according to the present invention. Preferably, the harvestable parts comprise the PPO nucleic acid or PPO protein of the present invention. The harvestable parts may be seeds, roots, leaves and/or flowers comprising the PPO nucleic acid or PPO protein or parts thereof. Preferred parts of soy plants are soy beans comprising the PPO nucleic acid or PPO protein.
In another embodiment, the invention refers to products derived from a plant according to the present invention, parts thereof or harvestable parts thereof. A preferred plant product is fodder, seed meal, oil, or seed-treatment-coated seeds. Preferably, the meal and/or oil comprises the mutated PPO nucleic acids or PPO proteins of the present invention.
a) growing the plants of the invention or obtainable by the methods of invention and
b) producing said product from or by the plants of the invention and/or parts, e.g. seeds, of these plants.
In another embodiment, the invention refers to a method for the production of a product, which method comprises
a) growing the plants of the invention,
b) removing the harvestable parts as defined above from the plants and
c) producing said product from or by the harvestable parts of the invention.
In a further embodiment the method comprises the steps
The product may be produced at the site where the plant has been grown, the plants and/or parts thereof may be removed from the site where the plants have been grown to produce the product. Typically, the plant is grown, the desired harvestable parts are removed from the plant, if feasible in repeated cycles, and the product made from the harvestable parts of the plant. The step of growing the plant may be performed only once each time the methods of the invention is performed, while allowing repeated times the steps of product production e.g. by repeated removal of harvestable parts of the plants of the invention and if necessary further processing of these parts to arrive at the product. It is also possible that the step of growing the plants of the invention is repeated and plants or harvestable parts are stored until the production of the product is then performed once for the accumulated plants or plant parts. Also, the steps of growing the plants and producing the product may be performed with an overlap in time, even simultaneously to a large extend or sequentially. Generally the plants are grown for some time before the product is produced.
In one embodiment the products produced by said methods of the invention are plant products such as, but not limited to, a foodstuff, feedstuff, a food supplement, feed supplement, fiber, cosmetic and/or pharmaceutical Foodstuffs are regarded as compositions used for nutrition and/or for supplementing nutrition. Animal feedstuffs and animal feed supplements, in particular, are regarded as foodstuffs.
In another embodiment the inventive methods for the production are used to make agricultural products such as, but not limited to, plant extracts, proteins, amino acids, carbohydrates, fats, oils, polymers, vitamins, and the like.
It is possible that a plant product consists of one or more agricultural products to a large extent.
−1
−1
−1
−1
As described above, the present invention teaches compositions and methods for increasing the PPO-inhibiting tolerance of a crop plant or seed as compared to a wild-type variety of the plant or seed. In a preferred embodiment, the PPO-inhibiting tolerance of a crop plant or seed is increased such that the plant or seed can withstand a uracilpyridine herbicide application of preferably approximately 1-1000 g ai ha, more preferably 1-200 g ai ha, even more preferably 5-150 g ai ha, and most preferably 10-100 g ai ha. As used herein, to “withstand” a uracilpyridine herbicide application means that the plant is either not killed or only moderately injured by such application. It will be understood by the person skilled in the art that the application rates may vary, depending on the environmental conditions such as temperature or humidity, and depending on the chosen kind of herbicide (active ingredient ai).
Furthermore, the present invention provides methods that involve the use of at least one uracilpyridine herbicide, optionally in combination with one or more herbicidal compounds B, and, optionally, a safener C, as described in detail supra.
In these methods, the uracilpyridine herbicide can be applied by any method known in the art including, but not limited to, seed treatment, soil treatment, and foliar treatment. Prior to application, the uracilpyridine herbicide can be converted into the customary formulations, for example solutions, emulsions, suspensions, dusts, powders, pastes and granules. The use form depends on the particular intended purpose; in each case, it should ensure a fine and even distribution of the compound according to the invention.
By providing plants having increased tolerance to uracilpyridine herbicide, a wide variety of formulations can be employed for protecting plants from weeds, so as to enhance plant growth and reduce competition for nutrients. A uracilpyridine herbicide can be used by itself for pre-emergence, post-emergence, pre-planting, and at-planting control of weeds in areas surrounding the crop plants described herein, or a uracilpyridine herbicide formulation can be used that contains other additives. The uracilpyridine herbicide can also be used as a seed treatment. Additives found in a uracilpyridine herbicide formulation include other herbicides, detergents, adjuvants, spreading agents, sticking agents, stabilizing agents, or the like. The uracilpyridine herbicide formulation can be a wet or dry preparation and can include, but is not limited to, flowable powders, emulsifiable concentrates, and liquid concentrates. The uracilpyridine herbicide and herbicide formulations can be applied in accordance with conventional methods, for example, by spraying, irrigation, dusting, or the like.
Suitable formulations are described in detail in PCT/EP2009/063387 and PCT/EP2009/063386, which are incorporated herein by reference.
It should also be understood that the foregoing relates to preferred embodiments of the present invention and that numerous changes may be made therein without departing from the scope of the invention. The invention is further illustrated by the following examples, which are not to be construed in any way as imposing limitations upon the scope thereof.
On the contrary, it is to be clearly understood that resort may be had to various other embodiments, modifications, and equivalents thereof, which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the present invention and/or the scope of the appended claims.
EXAMPLES
Example 1: Site-Directed Mutagenesis PPO
Example 2: Expression and Purification of Recombinant Wildtype and Mutant PPO
Example 3: PPO Enzyme Assay (Non-Recombinant)
Example 4: PPO Enzyme Assay (Recombinant)
Example 5: Engineering PPO-Derivative Herbicide Tolerant Plants Having Wildtype or Mutated PPO Sequences
Example 6: Tissue Culture Conditions
Example 7: Selection of Herbicide-Tolerant Calli
Example 8: Regeneration of Plants
Example 9: Sequence Analysis
Example 10: Demonstration of Herbicide-Tolerance
Example 11: Demonstration of Herbicide Tolerance in Mutagenized Non-Transgenic Rice
Example 12: Herbicide Selection Using Tissue Culture
Example 13: Maize Whole Plant Transformation and PPO Inhibitor Tolerance Testing
Example 14: Soybean Transformation and PPO Inhibitor Tolerance Testing
Uracilpyridine 1
ethyl 2-[[3-[2-chloro-4-fluoro-5-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-
yl]phenoxy]-2-pyridyl]oxy]acetate
Uracilpyridine 2
ethyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 3
2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]-2-pyridyl]oxy]acetic acid
Uracilpyridine 4
ethyl 2-[2-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetate
Uracilpyridine 5
2-[2-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetic acid
Uracilpyridine 6
ethyl 2-[2-[[3-bromo-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetate
Uracilpyridine 7
ethyl 2-[2-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]-4-fluoro-phenoxy]acetate
Uracilpyridine 8
ethyl 2-[2-[[3,5-difluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetate
Uracilpyridine 9
2-[2-[[3,5-difluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetic acid
Uracilpyridine 10
2-[2-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]-N-methylsulfonyl-acetamide
Uracilpyridine 11
ethyl 2-[[3-[[3-chloro-6-[3,5-dimethyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-5-
fluoro-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 12
ethyl 2-[2-[[3-chloro-6-[3,5-dimethyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-5-fluoro-
2-pyridyl]oxy]phenoxy]acetate
Uracilpyridine 13
allyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 14
prop-2-ynyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-
1-yl]-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 15
cyclopropylmethyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-
(trifluoromethyl)pyrimidin-1-yl]-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 16
2,2-difluoroethyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-
(trifluoromethyl)pyrimidin-1-yl]-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 17
isobutyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-
2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 18
(2-ethoxy-2-oxo-ethyl) 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-
(trifluoromethyl)pyrimidin-1-yl]-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 19
2-methoxyethyl 2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-
(trifluoromethyl)pyrimidin-1-yl]-2-pyridyl]oxy]-2-pyridyl]oxy]acetate
Uracilpyridine 20
2-[[3-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]-2-pyridyl]oxy]-N-methylsulfonyl-acetamide
Uracilpyridine 21
methyl 2-[2-[[3-chloro-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-
2-pyridyl]oxy]phenoxy]acetate
Uracilpyridine 22
ethyl 2-[2-[[3-bromo-5-fluoro-6-[3-methyl-2,6-dioxo-4-(trifluoromethyl)pyrimidin-1-yl]-2-
pyridyl]oxy]phenoxy]acetate
E. coli
All nucleic acid coding sequence and all single and double mutants encoding a herbicide tolerant PPO polypeptides are synthesized and cloned by Geneart (Geneart AG, Regensburg, Germany). Rational design mutants are synthesized by Geneart. Random PPO gene libraries are synthesized by Geneart. Plasmids are isolated from TOP10 by performing a plasmid minpreparation and confirmed by DNA sequencing.
E. coli
(Taken from: Franck E. Dayan, Pankaj R. Daga, Stephen O. Duke, Ryan M. Lee, Patrick J. Tranel, Robert J. Doerksen. Biochemical and structural consequences of aglycine deletion in the α-8 helix of protoporphyrinogen oxidase. Biochimica et Biophysica Acta 1804 (2010), 1548-56) Clones in pRSET vector are transformed into BL21(DE3)-pLysS strain of . Cells are grown in 250 mL of LB with 100 μgmL-1 of carbenicillin, shaking overnight at 37° C. Cultures are diluted in 1 L of LB with antibiotic and grown at 37° C. shaking for 2 h, induced with 1 mM IPTG and grown at 25° C. shaking for 5 more hours. The cells are harvested by centrifugation at 1600×g, washed with 0.09% NaCl, and stored at −80° C. Cells are lysed using a French press at 140 MPa in 50 mM sodium phosphate pH 7.5, 1 M NaCl, 5 mM imidazole, 5% glycerol, and 1 μg mL-1 leupeptin. Following lysis, 0.5 U of benzonase (Novagen, EMD Chemicals, Inc., Gibbstown, N.J.) and PMSF (final concentration of 1 mM) are added. Cell debris is removed by centrifugation at 3000×g. His-tagged PPO proteins are purified on a nickel activated Hitrap Chelating HP column (GE Healthcare Bio-Sciences Corp., Piscataway, N.J.) equilibrated with 20 mM sodium phosphate pH 8.0, 50 mM NaCl, 5 mM imidazole, 5 mM MgCl2, 0.1 mM EDTA, and 17% glycerol. PPO is eluted with 250 mM imidazole. The active protein is desalted on a PD-10 column (GE Healthcare Bio-Sciences Corp., Piscataway, N.J.) equilibrated with a 20 mM sodium phosphate buffer, pH 7.5, 5 mM MgCl2, 1 mM EDTA and 17% glycerol. Each litre of culture provided approximately 10 mg of pure PPO, which is stored at −20° C. until being used in assays.
−1
−1
50
PPO protein (EC 1.3.3.4) is extracted from coleoptiles or shoots (150 g fresh weight) of dark-grown corn, black nightshade, morning glory, and velvetleaf seedlings as described previously (Grossmann et al. 2010). Before harvesting, the seedlings are allowed to green for 2 hours in the light in order to achieve the highest specific enzyme activities in the thylakoid fractions at low chlorophyll concentrations. At high chlorophyll concentrations significant quenching of fluorescence occurs, which limits the amount of green thylakoids that can be used in the test. Plant materials are homogenized in the cold with a Braun blender using a fresh-weight-to-volume ratio of 1:4. Homogenization buffer consisted of tris(hydroxymethyl)aminomethane (Tris)-HCl (50 mM; pH 7.3), sucrose (0.5 M), magnesium chloride (1 mM), ethylenediaminetetraacetic acid (EDTA) (1 mM) and bovine serum albumin (2 g L). After filtration through four layers of Miracloth, crude plastid preparations are obtained after centrifugation at 10 000×g for 5 min and resuspension in homogenization buffer before centrifugation at 150×g for 2 min to remove crude cell debris. The supernatant is centrifuged at 4000×g for 15 min and the pellet fraction is resuspended in 1 ml of a buffer containing Tris-HCl (50 mM; pH 7.3), EDTA (2 mM), leupeptin (2 μM), pepstatin (2 μM) and glycerol (200 ml L) and stored at −80° C. until use. Protein is determined in the enzyme extract with bovine serum albumin as a standard. PPO activity is assayed fluorometrically by monitoring the rate of Proto formation from chemically reduced protoporphyrinogen IX under initial velocity conditions. The assay mixture consisted of Tris-HCl (100 mM; pH 7.3), EDTA (1 mM), dithiothreitol (5 mM), Tween 80 (0.085%), protoporphyrinogen IX (2 μM), and 40 μg extracted protein in a total volume of 200 μl. The reaction is initiated by addition of substrate protoporphyrinogen IX at 22° C. The uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control are prepared in dimethyl sulfoxide (DMSO) solution (0.1 mM concentration of DMSO in the assay) and added to the assay mixture in concentrations of 0.005 μM to 5 μM before incubation. Fluorescence is monitored directly from the assay mixture using a POLARstar Optima/Galaxy (BMG) with excitation at 405 nm and emission monitored at 630 nm. Non-enzymatic activity in the presence of heat-inactivated extract is negligible. Inhibition of enzyme activity induced by the herbicide is expressed as percentage inhibition relative to untreated controls. Molar concentrations of compound required for 50% enzyme inhibition (ICvalues) are calculated by fitting the values to the dose-response equation using non-linear regression analysis.
50
Proto is purchased from Sigma-Aldrich (Milwaukee, Wis.). Protogen is prepared according to Jacobs and Jacobs (N.J. Jacobs, J. M. Jacobs, Assay for enzymatic protoporphyrinogen oxidation, a late step in heme synthesis, Enzyme 28 (1982) 206-219). Assays are conducted in 100 mM sodium phosphate pH 7.4 with 0.1 mM EDTA, 0.1% Tween 20, 5 μM FAD, and 500 mM imidazole. Dose-response curves with the uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control, and MC-15608 are obtained in the presence of 150 μM Protogen. The excitation and emission bandwidths are set at 1.5 and 30 nm, respectively. All assays are made in duplicates or triplicates and measured using a POLARstar Optima/Galaxy (BMG) with excitation at 405 nm and emission monitored at 630 nm. Molar concentrations of compound required for 50% enzyme inhibition (ICvalues) are calculated by fitting the values to the dose-response equation using non-linear regression analysis. The results are shown in Table x.
TABLE x
Variant PPO
Enzyme and IC50 (M)
Wild Type
Tolerance
Based on SEQ ID NO: 1
PPO Enzyme
Factor
Compound
(Amaranthus PPO2)
IC50 (M)
(R/S)
Uracilpyridine
R128A, F420V
2.80E−07
1.77E−10
1579
1
Uracilpyridine
R128A, F420V
1.62E−07
1.77E−10
915
1
Uracilpyridine
R128A, F420M
6.16E−08
1.77E−10
347
1
Uracilpyridine
L397Q, F420V
2.26E−06
1.77E−10
12739
1
Uracilpyridine
L397Q, F420M
4.29E−07
1.77E−10
2418
1
Uracilpyridine
L397Q
2.38E−09
1.77E−10
13
1
Uracilpyridine
F420V
6.64E−08
1.77E−10
374
1
Uracilpyridine
F420M
3.81E−08
1.77E−10
215
1
Uracilpyridine
F420M
4.69E−08
1.77E−10
265
1
Uracilpyridine
R128A, F420V
2.97E−07
1.73E−10
1718
4
Uracilpyridine
R128A, F420M
6.94E−08
1.73E−10
401
4
Uracilpyridine
F420M
9.44E−09
1.73E−10
55
4
Uracilpyridine
R128A, F420V
1.49E−07
1.87E−10
796
2
Uracilpyridine
R128A, F420V
2.07E−07
1.87E−10
1105
2
Uracilpyridine
R128A, F420M
9.74E−08
1.87E−10
521
2
Uracilpyridine
L397Q, F420V
2.56E−06
1.87E−10
13678
2
Uracilpyridine
L397Q, F420M
9.05E−07
1.87E−10
4843
2
Uracilpyridine
L397Q
1.29E−09
1.87E−10
7
2
Uracilpyridine
F420V
1.06E−07
1.87E−10
567
2
Uracilpyridine
F420M
4.57E−08
1.87E−10
244
2
Uracilpyridine
F420M
7.76E−08
1.87E−10
415
2
Uracilpyridine
R128A, F420V
2.25E−06
4.96E−10
4531
8
Uracilpyridine
F420M
6.91E−08
4.96E−10
139
8
Uracilpyridine
R128A, F420M
1.67E−06
2.12E−10
7879
10
Uracilpyridine
L397E, F420M
6.51E−06
2.12E−10
30714
10
Uracilpyridine
R128A, F420M
1.14E−07
2.12E−10
536
12
Uracilpyridine
R128A, F420M
9.18E−08
2.18E−10
421
13
Uracilpyridine
R128A, F420M
1.58E−07
2.34E−10
675
14
Uracilpyridine
R128A, F420M
1.25E−07
2.48E−10
503
19
Uracilpyridine
R128A, F420M
4.29E−08
2.20E−10
195
15
Uracilpyridine
R128A, F420M
8.90E−08
1.75E−10
508
16
Uracilpyridine
R128A, F420M
4.61E−08
1.13E−10
406
17
Uracilpyridine
R128A, F420M
1.16E−07
1.54E−10
753
18
Glyceine max
Arabidopsis thaliana
Agrobacterium tumefaciens
Agrobacterium
Agrobacterium
PPO-derivative herbicide tolerant soybean (), plants are produced by a method as described by Olhoft et al. (US patent 2009/0049567). For transformation of soybean or , Wildtype or Mutated PPO sequences encoding herbicide tolerant PPO polypeptides are cloned with standard cloning techniques as described in Sambrook eta. (Molecular cloning (2001) Cold Spring Harbor Laboratory Press) in a binary vector containing resistance marker gene cassette (AHAS) and mutated PPO sequence (marked as GOI) in between ubiquitin promoter (PcUbi) and nopaline synthase terminator (NOS) sequence. For corn transformation, Wildtype or Mutated PPO sequences are cloned with standard cloning techniques as described in Sambrook et a. (Molecular cloning (2001) Cold Spring Harbor Laboratory Press) in a binary vector containing resistance marker gene cassette (AHAS) and mutated PPO sequence (marked as GOI) in between corn ubiquitin promoter (ZmUbi) and nopaline synthase terminator (NOS) sequence. Binary plasmids are introduced to for plant transformation. Plasmid constructs are introduced into soybean's axillary meristem cells at the primary node of seedling explants via -mediated transformation. After inoculation and co-cultivation with Agrobacteria, the explants are transferred to shoot introduction media without selection for one week. The explants are subsequently transferred to a shoot induction medium with 1-3 μM imazapyr (Arsenal) for 3 weeks to select for transformed cells. Explants with healthy callus/shoot pads at the primary node are then transferred to shoot elongation medium containing 1-3 μM imazapyr until a shoot elongated or the explant died. Transgenic plantlets are rooted, subjected to TaqMan analysis for the presence of the transgene, transferred to soil and grown to maturity in greenhouse. Plant transformation vector constructs containing mutated PPO sequences are introduced into maize immature embryos via -mediated transformation according to the procedure outlined in Peng et al. (WO2006/136596).
Arabidopsis thaliana
Arabidopsis
Oryza sativa
Transformed cells are selected in selection media supplemented with 0.5-1.5 μM imazethapyr for 3-4 weeks. Transgenic plantlets are regenerated on plant regeneration media and rooted afterwards. Transgenic plantlets are subjected to TaqMan analysis for the presence of the transgene before being transplanted to potting mixture and grown to maturity in greenhouse. are transformed with wildtype or mutated PPO sequences by floral dip method as described by McElver and Singh (WO 2008/124495). Transgenic plants are subjected to TaqMan analysis for analysis of the number of integration loci. Transformation of (rice) are done by protoplast transformation as described by Peng et a. (U.S. Pat. No. 6,653,529) T0 or T1 transgenic plants of soybean, corn, and rice containing mutated PPO sequences are tested for improved tolerance to PPO-derived herbicides in greenhouse studies and mini-plot studies with the uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control.
An in vitro tissue culture mutagenesis assay has been developed to isolate and characterize plant tissue (e.g., maize, rice tissue) that is tolerant to protoporphyrinogen oxidase inhibiting herbicides, e.g. the uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control). The assay utilizes the somaclonal variation that is found in in vitro tissue culture. Spontaneous mutations derived from somaclonal variation can be enhanced by treatment with a chemical mutagen (e.g. Ethyl methanesulfonate, N-ethyl-N-nitrosourea, N-Nitroso-N-methylurea) and subsequent selection in a stepwise manner, on increasing concentrations of herbicide.
The present invention provides tissue culture conditions for encouraging growth of friable, embryogenic maize or rice callus that is regenerable. Calli are initiated from rice cultivar Indica (Indica I). Seeds are surface sterilized in 70% ethanol for approximately 1 min followed by 20% commercial Clorox bleach for 20 minutes. Seeds are rinsed with sterile water and plated on R001 M media. The ingredient lists for the media tested are presented in Table y.
TABLE y
Ingredients
Supplier
R001M
R025M
R026M
R327M
R008S
N6 Salts
Phytotech
4 g/L
4 g/L
4.3 g/L
4.3 g/L
3.25 g/L
N6 vitamins
Phytotech
1X
1X
1X
1X
Maltose
VWR
30 g/L
30 g/L
30 g/L
30 g/L
Casamino Acid
BD
0.3 g/L
0.3 g/L
2 g/L
Proline
Sigma
2.9 g/L
0.5 g/L
2,4-D
ICN
2 mg/L
MES
Sigma
0.5 g/L
0.5 g/L
0.5 g/L
0.5 g/L
0.5 g/L
MS Salts
Phytotech
MS Vitamins
Phytotech
Sorbitol
Sigma
30 g/L
Sucrose
Sigma
20 g/L
Nicotinic Acid
Sigma
0.5 mg/L
Pyridoxine HCL
Sigma
0.5 mg/L
Thiamine HCL
Sigma
1 mg/L
Myo-inositol
Sigma
0.1 g/L
R001M callus induction media is selected after testing numerous variations. Cultures are kept in the dark at 30° C. Embryogenic callus is subcultured to fresh media after 10-14 days.
Once tissue culture conditions are determined, further establishment of selection conditions are established through the analysis of tissue survival in kill curves with saflufenacil, trifludimoxazin, sulfentrazone, and the uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control. Careful consideration of accumulation of the herbicide in the tissue, as well as its persistence and stability in the cells and the culture media is performed. Through these experiments, a sub-lethal dose has been established for the initial selection of mutated material. After the establishment of the starting dose of uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control in selection media, the tissues are selected in a step-wise fashion by increasing the concentration of the PPO inhibitor with each transfer until cells are recovered that grew vigorously in the presence of toxic doses. The resulting calli are further subcultured every 3-4 weeks to R001M with selective agent. Over 26,000 calli are subjected to selection for 4-5 subcultures until the selective pressure is above toxic levels as determined by kill curves and observations of continued culture. Alternatively, liquid cultures initiated from calli in MS711R with slow shaking and weekly subcultures. Once liquid cultures are established, selection agent is added directly to the flask at each subculture. Following 2-4 rounds of liquid selection, cultures are transferred to filters on solid R001M media for further growth.
Oryza sative
Tolerant tissue, e.g. rice tissue, is regenerated and characterized molecularly for PPO gene, e.g. PPO2 sequence mutations and/or biochemically for altered PPO activity in the presence of the selective agent. In addition, genes involved directly and/or indirectly in tetrapyrrole biosynthesis and/or metabolism pathways are also sequenced to characterize mutations. Finally, enzymes that change the fate (e.g. metabolism, translocation, transportation) are also sequence to characterized mutations. Following herbicide selection, calli are regenerated using a media regime of R025M for 10-14 days, R026M for ca. 2 weeks, R327M until well formed shoots are developed, and R008S until shoots are well rooted for transfer to the greenhouse. Regeneration is carried out in the light. No selection agent is included during regeneration. Once strong roots are established, M0 regenerants are transplant to the greenhouse in square or round pots. Transplants are maintained under a clear plastic cup until they are adapted to greenhouse conditions. The greenhouse is set to a day/night cycle of 27° C./21° C. (80° F./70° F.) with 600 W high pressure sodium lights supplementing light to maintain a 14 hour day length. Plants are watered according to need, depending in the weather and fertilized daily.
Leaf tissue is collected from clonal plants separated for transplanting and analyzed as individuals. Genomic DNA is extracted using a Chloropure Nucleic acid extraction kit (Agencourt, U.S. Pat. Nos. 5,898,071; 5,705,628; 6,534,262) as directed by the manufacturer. Isolated DNA is PCR amplified using the appropriate forward and reverse primer. PCR amplification is performed using LongAmp HotStart Taq DNA Polymerase Mix (New England Biolabs) using thermocycling program as follows: 94° C. for 30 sec, followed by 35 cycles (94° C., 30 sec; 54° C., 30 sec; 65° C., 300 sec), 10 min at 65° C.
PCR products are verified for concentration and fragment size via agarose gel electrophoresis. Dephosphorylated PCR products are analyzed by direct sequence using the PCR primers (Genewiz or GenScript). Chromatogram trace files (.scf) are analyzed for mutation relative to the wild-type gene using Sequencher (Gene Codes) or Vector NTI Advance 10™ (Invitrogen). Based on sequence information, mutations are identified in several individuals. Sequence analysis is performed on the representative chromatograms and corresponding alignment with default settings and edited to call secondary peaks.
T0 or T1 transgenic plant of soybean, corn, Canola varieties and rice containing PPO1 and or PPO2 sequences are tested for improved tolerance to herbicides in greenhouse studies and mini-plot studies with the uracilpyridines disclosed SUPRA, and photosynthesis inhibitor diuron as negative control. For the pre-emergence treatment, the herbicides are applied directly after sowing by means of finely distributing nozzles. The containers are irrigated gently to promote germination and growth and subsequently covered with transparent plastic hoods until the plants have rooted. This cover causes uniform germination of the test plants, unless this has been impaired by the herbicides. For post emergence treatment, the test plants are first grown to a height of 3 to 15 cm, depending on the plant habit, and only then treated with the herbicides. For this purpose, the test plants are either sown directly, and grown in the same containers or they are first grown separately and transplanted into the test containers a few days prior to treatment.
For testing of T0 plants, cuttings can be used. In the case of soybean plants, an optimal shoot for cutting is about 7.5 to 10 cm tall, with at least two nodes present. Each cutting is taken from the original transformant (mother plant) and dipped into rooting hormone powder (indole-3-butyric acid, IBA). The cutting is then placed in oasis wedges inside a bio-dome. Wild type cuttings are also taken simultaneously to serve as controls. The cuttings are kept in the bio-dome for 5-7 days and then transplanted to pots and then acclimated in the growth chamber for two more days. Subsequently, the cuttings are transferred to the greenhouse, acclimated for approximately 4 days, and then subjected to spray tests as indicated. Depending on the species, the plants are kept at 10-25° C. or 20-35° C. The test period extends over 3 weeks. During this time, the plants are tended and their response to the individual treatments is evaluated. Herbicide injury evaluations are taken at 2 and 3 weeks after treatment. Plant injury is rated on a scale of 0% to 100%, 0% being no injury and 100% being complete death.
Arabidopsis thaliana
Physiologia Plantarum
−2
−1
Transgenic plants are assayed for improved tolerance to the uracilpyridines disclosed SUPRA, in 48-well plates. Therefore, T2 seeds are surface sterilized by stirring for 5 min in ethanol+water (70+30 by volume), rinsing one time with ethanol+water (70+30 by volume) and two times with sterile, deionized water. The seeds are resuspended in 0.1% agar dissolved in water (w/v) Four to five seeds per well are plated on solid nutrient medium consisting of half-strength murashige skoog nutrient solution, pH 5.8 (Murashige and Skoog (1962) 15: 473-497). Compounds are dissolved in dimethylsulfoxid (DMSO) and added to the medium prior solidification (final DMSO concentration 0.1%). Multi well plates are incubated in a growth chamber at 22° C., 75% relative humidity and 110 μmol Phot*m*swith 14: 10 h light: dark photoperiod. Growth inhibition is evaluated seven to ten days after seeding in comparison to wild type plants. The results are shown in Table i)
Compound
Gene Species Source
Construct Name
Tolerance Factor*
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_WT
6
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_F420M
110
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_L397D
50
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_L397D_F420V
41
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_R128A_F420M
50
Uracilpyridine 1
<i>Amaranthus tuberculatus</i>
AMATU_PPO2_R128A_F420V
75
Uracilpyridine 1
<i>Arabidopsis thaliana</i>
ARBTH_PPO1_WT
50
Uracilpyridine 1
<i>Arabidopsis thaliana</i>
ARBTH_PPO1_S305L_Y426M
250
Uracilpyridine 3
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
1600
Uracilpyridine 9
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
1600
Uracilpyridine 4
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
810000
Uracilpyridine 5
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
1400
Uracilpyridine 2
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
6400
Uracilpyridine 8
<i>Escherichia coli</i>
AMATU_PPO2_TP_hemG
8202
Transgenic T2 <i>Arabidopsis</i> plants harboring PPO inhibitor tolerance trait in lab based germination assay.
Arabidopsi
FIG. 1
Additionally, transgenic T2 plants are tested for improved tolerance to herbicides in greenhouse studies with the uracilpyridines as disclosed SUPRA, and photosynthesis inhibitor diuron as negative control. Results are shown in table ii), table iii), and .
TABLE ii
Transgenic T2 Arabidopsis plants, sprayed post in the greenhouse with the indicated
amount of PPO inhibitor + 1% (v/v) MSO. Evaluation performed 14 Days After Treatment (DAT)
and is shown as injury (%) relative to non-transgenic treated plants.
ARBTH WT MC24 - non-transgenic
ARBTH_PPO1_WT
AMATU_PPO2_WT
ALOMY_PPO2_TP_AMATU_PPO2_R128M_F420I
AMATU_PPO2_R128M_F420I
Compound
gai/ha
/
Q
Q
R
K
Uracilpyridine 2
200
100
75
70
100
100
45
58
80
100
50
100
45
18
30
100
100
100
10
100
100
100
5
100
100
100
1
100
15
23
Uracilpyridine 3
80
100
30
100
100
100
10
100
98
100
5
100
83
80
1
88
10
48
Uracilpyridine 4
200
100
80
73
100
100
60
58
80
100
50
100
40
30
30
100
100
100
10
100
100
100
5
100
99
88
1
100
33
25
Uracilpyridine 5
80
100
30
100
100
100
10
100
99
100
5
100
90
90
1
100
45
28
Uracilpyridine 1
200
100
84
75
100
100
92
73
50
100
60
60
Uracilpyridine 10
200
100
50
Uracilpyridine 6
200
100
50
Uracilpyridine 7
200
100
50
Uracilpyridine 8
80
100
30
100
100
100
10
100
55
100
5
100
30
55
1
88
10
10
Uracilpyridine 9
80
100
30
100
98
100
10
100
70
100
5
100
50
68
1
85
10
38
Uracilpyridine 10
80
100
30
100
100
100
10
100
100
100
5
100
98
100
1
100
50
58
AMATU_PPO2_R128A_F420M
AMATU_PPO2_TP_hemG
AMATU_PPO2_TP_hemG
ALOMY_PPO2_R137L_F438V
ALOMY_PPO2_R137L_F438V
Compound
gai/ha
Q
R
R
F
G
Uracilpyridine 2
200
65
35
30
23
20
100
15
40
33
18
10
80
50
10
55
28
18
0
30
10
5
1
Uracilpyridine 3
80
30
10
5
1
Uracilpyridine 4
200
60
65
20
8
100
60
38
20
15
80
50
55
30
8
8
30
10
5
1
Uracilpyridine 5
80
30
10
5
1
Uracilpyridine 1
200
75
53
65
28
100
60
63
33
75
50
100
55
60
20
5
Uracilpyridine 10
200
10
5
100
8
0
50
0
0
Uracilpyridine 6
200
18
0
100
0
0
50
0
0
Uracilpyridine 7
200
13
8
100
18
0
50
0
0
Uracilpyridine 8
80
30
10
5
1
Uracilpyridine 9
80
30
10
5
1
Uracilpyridine 10
80
30
10
5
1
AMATU_PPO2_R128A_F420V
AMATU_PPO2_R128A_F420V
AMATU_PPO2_L397E_F420V
AMATU_PPO2_L397E_F420V
ARBTH_PPO1_S305L_Y426M
Compound
gai/ha
A
D
D
L
O
Uracilpyridine 2
200
100
80
45
50
30
33
10
0
5
0
1
Uracilpyridine 3
80
13
30
49
10
0
5
0
1
Uracilpyridine 4
200
100
80
95
50
30
0
10
0
5
0
1
Uracilpyridine 5
80
73
30
68
10
0
5
0
1
Uracilpyridine 1
200
100
50
100
Uracilpyridine 10
200
68
88
13
20
100
53
78
5
53
50
38
30
0
8
Uracilpyridine 6
200
30
28
8
15
100
43
38
0
0
50
0
0
0
0
Uracilpyridine 7
200
70
75
10
33
100
50
65
0
0
50
13
33
0
0
Uracilpyridine 8
80
0
30
0
10
0
5
0
1
Uracilpyridine 9
80
0
30
0
10
0
5
0
1
Uracilpyridine 10
80
99
30
99
10
8
5
10
1
TABLE iii
Transgenic T2 Arabidopsis plants, sprayed post in the greenhouse with
the indicated amount of PPO inhibitor + 1% (v/v) MSO. Evaluation performed
14 Days After Treatment (DAT) and is shown as injury (%) relative to non-transgenic treated plants.
Compound
g ai/ha
ARBTH WT MC 24 (non-transgenic)
ALOMY_PPO2_R137L_F438M
ALOMY_PPO2_R137L_F438M
Uracilpyridine
100
100
0
0
2
50
100
0
0
Uracilpyridine
100
100
0
0
4
50
100
0
0
Uracilpyridine
100
100
20
0
1
50
100
0
0
Compound
g ai/ha
ALOMY_PPO2_R137L_F438M
ALOMY_PPO2_R137L_F438M
ALOMY_PPO2_R137L_F438V
Uracilpyridine
100
0
0
0
2
50
0
0
0
Uracilpyridine
100
0
0
0
4
50
0
0
10
Uracilpyridine
100
0
0
0
1
50
0
0
5
Compound
g ai/ha
ALOMY_PPO2_R137L_F438V
ALOMY_PPO2_R137L_F438V
ALOMY_PPO2_R137L_F438V
Uracilpyridine
100
0
0
0
2
50
0
0
0
Uracilpyridine
100
10
10
10
4
50
0
0
0
Uracilpyridine
100
25
35
40
1
50
50
15
0
Compound
g ai/ha
AMATU_PPO2_L397E_F420V
AMATU_PPO2_L397E_F420V
AMATU_PPO2_L397E_F420V
Uracilpyridine
100
0
0
0
2
50
0
0
0
Uracilpyridine
100
0
30
0
4
50
0
10
0
Uracilpyridine
100
0
35
55
1
50
0
10
0
Compound
g ai/ha
AMATU_PPO2_L397E_F420V
AMATU_PPO2_L397E_F420M
AMATU_PPO2_L397E_F420M
Uracilpyridine
100
0
10
35
2
50
15
0
0
Uracilpyridine
100
0
15
35
4
50
10
0
25
Uracilpyridine
100
0
15
30
1
50
35
0
10
Compound
g ai/ha
AMATU_PPO2_L397E_F420M
AMATU_PPO2_L397E_F420M
AMATU_PPO2_L397Q_F420V
Uracilpyridine
100
0
20
80
2
50
0
0
45
Uracilpyridine
100
60
20
80
4
50
0
0
60
Uracilpyridine
100
60
55
85
1
50
15
40
40
Compound
g ai/ha
ARBTH WT MC 24 (non-transgenic)
AMATU_PPO2_L397Q_F420V
AMATU_PPO2_R128A_F420L
Uracilpyridine
100
100
65
60
2
50
100
0
0
Uracilpyridine
100
100
—
10
4
50
100
—
0
Uracilpyridine
100
100
65
60
1
50
100
75
65
Compound
g ai/ha
AMATU_PPO2_R128A_F420L
AMATU_PPO2_R128A_F420L
AMATU_PPO2_R128A_F420L
Uracilpyridine
100
35
15
30
2
50
0
0
15
Uracilpyridine
100
65
20
20
4
50
10
0
0
Uracilpyridine
100
90
95
80
1
50
75
5
75
Compound
g ai/ha
AMATU_PPO2_R128A_F420I
AMATU_PPO2_R128A_F420I
AMATU_PPO2_R128A_F420I
Uracilpyridine
100
65
65
65
2
50
0
45
65
Uracilpyridine
100
65
50
55
4
50
0
50
40
Uracilpyridine
100
65
80
95
1
50
70
75
60
Compound
g ai/ha
AMATU_PPO2_R128A_F420I
AMATU_PPO2_R128A_F420M
AMATU_PPO2_R128A_F420M
Uracilpyridine
100
30
0
65
2
50
0
0
0
Uracilpyridine
100
65
25
65
4
50
50
0
15
Uracilpyridine
100
60
35
80
1
50
60
55
65
Compound
g ai/ha
AMATU_PPO2_R128A_F420M
AMATU_PPO2_R128A_F420M
AMATU_PPO2_R128A F420V
Uracilpyridine
100
30
0
85
2
50
30
0
0
Uracilpyridine
100
40
45
15
4
50
25
30
75
Uracilpyridine
100
65
65
95
1
50
25
40
90
Compound
g ai/ha
AMATU_PPO2_R128A F420V
AMATU_PPO2_R128A F420V
AMATU_PPO2_R128A F420V
Uracilpyridine
100
70
25
65
2
50
0
20
0
Uracilpyridine
100
75
25
40
4
50
70
60
75
Uracilpyridine
100
98
95
80
1
50
55
65
95
FIGS. 1 to 4
Selected mutants are transferred to pots and grown in the greenhouse for seed production. The resulting M1 progeny are screened for the presence and zygosity of tolerance conferring mutations in the OsPPO2 gene, e.g. mutated OsPPO2 polypeptides as shown in SEQ ID Nos: 628, 629, 630, 631, 632, 633, 634, 635, 636, or 637. M1 plants containing a homozygous in the OsPPO2 gene are kept for seed production to generate a stable, homozygous, herbicide tolerant line. Homozygous lines ca. 2 weeks old are sprayed using a track sprayer with uracilpyridine 2 or uracilpyridine 4 supplemented with 0.1% methylated seed oil. Once sprayed, plants are kept on drought conditions for 24 hours before being watered and fertilized again. Sprayed plants are photographed and rated for herbicide injury at 3 days and 7 days treatment. The results are shown in .
Media is selected for use and kill curves developed as specified above. For selection, different techniques are utilized. Either a step wise selection is applied, or an immediate lethal level of herbicide is applied. In either case, all of the calli are transferred for each new round of selection. Selection is 4-5 cycles of culture with 3-5 weeks for each cycle. Cali are placed onto nylon membranes to facilitate transfer (200 micron pore sheets, Biodesign, Saco, Me.). Membranes are cut to fit 100×20 mm Petri dishes and are autoclaved prior to use 25-35 calli (average weight/calli being 22 mg) are utilized in every plate. In addition, one set of calli are subjected to selection in liquid culture media with weekly subcultures followed by further selection on semi-solid media. Mutant lines are selected using the uracilpyridines as disclosed SUPRA, and photosynthesis inhibitor diuron as negative control. Efficiencies of obtaining mutants is high either based on a percentage of calli that gave rise to a regenerable, mutant line or the number of lines as determined by the gram of tissue utilized.
Immature embryos are transformed according to the procedure outlined in Peng et al. (WO2006/136596). Plants are tested for the presence of the T-DNA by Taqman analysis with the target being the nos terminator which is present in all constructs. Healthy looking plants are sent to the greenhouse for hardening and subsequent spray testing. The plants are individually transplanted into MetroMix 360 soil in 4″ pots. Once in the greenhouse (day/night cycle of 27° C./21° C. with 14 hour day length supported by 600 W high pressure sodium lights), they are allowed to grow for 14 days. T0 or T1 plants are sprayed with a treatment of the uracilpyridines as disclosed SUPRA and photosynthesis inhibitor diuron as negative control. Herbicide injury evaluations are taken at 7, 14 and 21 days after treatment. Herbicide injury evaluations are taken 2, 7, 14 and 21 days post-spray to look for injury to new growth points and overall plant health. The top survivors are transplanted into gallon pots filled with MetroMix 360 for seed production.
FIGS. 7 and 8
Transgenic T2 Corn plants harbouring PPO inhibitor tolerance trait, were sprayed post with the indicated amount of uracilpyridine+1% (v/v) MSO. Evaluation performed 14 Days After Treatment (DAT) and shown as injury (0-9) relative to non-transgenic treated plants. Injury Scale: 0-9, 0=no injury, 9=maximum injury. Note: segregating lines. The results are shown in the following Table and in .
Corn Trial
R128A,
R128A,
R128A,
J553
Rate
F420I
F420L
F420L
non
Compound
(g/ha)
Event 1
Event 2
Event 3
transgenic
Check
0
0
1
5
1
1
1
6
1
2
0
6
1% MSO
0
0
0
1
1
0
0
2
1
1
1
1
Uracilpyridine
25
1
0
1
9
2
0
1
0
9
1
0
0
9
Uracilpyridine
50
0
1
0
9
2
1
1
0
9
0
1
1
9
Uracilpyridine
100
1
1
0
9
2
0
1
1
9
1
1
1
9
Uracilpyridine
25
0
0
0
9
4
0
0
1
7
0
0
0
7
Uracilpyridine
50
0
1
1
9
4
1
0
0
7
0
1
0
9
Uracilpyridine
100
1
0
1
9
4
0
1
0
9
0
1
0
9
Transgenic T3Maize plants harboring a PPO inhibitor tolerance trait sprayed post in the field at the V3 leaf stage with 100 g/ha of PPO inh. 2+1% (v/v) MSO. Evaluation performed 2 and 12 Days After Treatment (DAT) and is shown as injury (%) relative to non-transgenic treated plants. Pictures taken 2 DAT.
Plant Injury (%)
J553/TR5753
Non-
AMATU_PPO2_
AMATU_PPO2_
DAT
transgenic
R128A_F420L
R128A_F420I
2
95
0
0
12
100
0
0
Soybean cv Jake is transformed as previously described by Siminszky et al., Phytochem Rev. 5:445-458 (2006). After regeneration, transformants are transplanted to soil in small pots, placed in growth chambers (16 hr day/8 hr night; 25° C. day/23° C. night; 65% relative humidity; 130-150 microE m-2 s-1) and subsequently tested for the presence of the T-DNA via Taqman analysis. After a few weeks, healthy, transgenic positive, single copy events are transplanted to larger pots and allowed to grow in the growth chamber. An optimal shoot for cutting is about 3-4 inches tall, with at least two nodes present. Each cutting is taken from the original transformant (mother plant) and dipped into rooting hormone powder (indole-3-butyric acid, IBA). The cutting is then placed in oasis wedges inside a bio-dome. The mother plant is taken to maturity in the greenhouse and harvested for seed. Wild type cuttings are also taken simultaneously to serve as negative controls. The cuttings are kept in the bio-dome for 5-7 days and then transplanted to 3 inch pots and then acclimated in the growth chamber for two more days. Subsequently, the cuttings are transferred to the greenhouse, acclimated for approximately 4 days prior to spray. T0 or 2-week old T1 plants are sprayed with a treatment of the uracilpyridines as disclosed SUPRA and photosynthesis inhibitor diuron as negative control. Herbicide injury evaluations are taken at 2, 7, 14 and 21 days after treatment.
Transgenic T2 or T3 Soybean plants harbouring PPO inhibitor tolerance trait, were sprayed post in the greenhouse with the indicated amount of uracilpyridine+1% (v/v) MSO. Evaluation performed 7 Days After Treatment (DAT) and shown as injury (0-9) relative to non-transgenic treated plants. Injury Scale: 0-9, 0=no injury, 9=maximum injury. Note: segregating lines.
R128A,
R128A,
L397E,
L397E,
L397Q,
Jake
Rate
F420L
F420I
F420V
F420M
F420M
non
Compound
(g/ha)
Event 1
Event 2
Event 3
Event 4
Event 5
transgenic
Uracilpyridine
25
1
1
2
1
1
9
1
1
1
1
2
2
9
1
*
1
1
2
9
1
9
1
1
1
9
Uracilpyridine
50
1
1
9
2
1
9
1
1
*
1
0
2
9
1
*
1
1
1
9
1
3
2
1
2
9
Uracilpyridine
100
1
1
1
1
3
9
1
1
2
1
2
2
9
1
*
9
*
*
9
4
1
9
1
3
9
Uracilpyridine
200
1
1
9
4
3
9
1
1
1
2
1
3
9
1
1
9
2
*
9
1
1
1
2
4
9
1% MSO
1
*
1
1
1
3
1
*
2
1
2
2
1
1
1
2
2
2
2
*
1
1
1
3
Check
1
*
0
1
0
1
0
0
1
1
2
0
0
*
*
1
*
1
1
*
2
1
*
1
FIGS. 7 and 8
Transgenic T2 or T3 Soybean plants harbouring PPO inhibitor tolerance trait, were sprayed post with the indicated amount of uracilpyridine+1% (v/v) MSO. Evaluation performed 14 Days After Treatment (DAT) and shown as injury (0-9) relative to non-transgenic treated plants. Injury Scale: 0-9, 0=no injury, 9=maximum injury. Note: segregating lines. The results are shown in the following Table and in .
Soybean Trial
Rate
L397E, F420V
L397E, F420M
L397Q, F420M
Jake
Compound
(g/ha)
Event 1
Event 2
Event 3
non transgenic
Check
0
9
9
9
0
0
2
9
0
2
1
9
1% MSO
0
0
0
1
0
0
0
1
0
0
0
1
Uracilpyridine
25
0
0
0
9
2
0
0
9
9
0
0
3
9
Uracilpyridine
50
0
0
3
9
2
0
0
9
9
0
0
1
9
Uracilpyridine
100
0
0
1
9
2
0
0
3
9
0
0
2
9
Uracilpyridine
25
0
0
1
9
4
0
0
9
9
0
0
0
9
Uracilpyridine
50
0
0
9
9
4
0
0
9
9
0
0
3
9
Uracilpyridine
100
0
0
3
9
4
0
0
4
9
0
1
3
9
FIGS. 9 through 12
Transgenic T4 Soybean plants harbouring PPO inhibitor tolerance trait, were sprayed post with the indicated amount of uracilpyridine+1% (v/v) MSO. Evaluation performed 7 Days After Treatment (DAT) and shown as injury (0-9) relative to non-transgenic treated plants. Injury Scale: 0-9, 0=no injury, 9=maximum injury. Asterisks indicates plants that did not germinate. Results are also shown in .
R128A +
R128A +
L397E +
L397E +
L397Q +
Jake
F420L
F420I
F420V
F420M
F420M
non-
Compound
Rate (g/ha)
Event 1
Event 2
Event 3
Event 4
Event 5
transgenic
Uracilpyridine 10
25
0
1
1
0
3
9
1
0
1
1
*
9
0
2
0
0
3
9
0
1
0
0
5
9
1
1
*
0
*
*
0
1
*
0
3
9
Uracilpyridine 10
50
1
1
*
0
4
9
1
2
1
1
4
9
0
1
*
0
4
9
1
2
*
0
4
9
0
2
0
0
4
9
1
2
1
1
4
9
Uracilpyridine 10
100
2
2
*
1
9
1
2
0
2
4
9
2
2
0
1
*
9
2
3
0
1
5
9
2
3
1
0
5
9
2
3
*
1
*
9
Uracilpyridine 20
25
2
2
0
0
4
9
0
2
1
0
3
9
1
2
0
1
*
9
0
2
*
0
3
9
1
2
0
0
*
9
2
1
*
0
3
9
Uracilpyridine 20
50
3
3
0
0
*
9
3
3
0
1
*
9
2
3
0
0
3
9
1
2
1
1
4
9
2
3
1
0
4
9
2
3
0
0
4
9
Uracilpyridine 20
100
3
4
*
0
*
9
4
4
0
0
4
9
5
4
0
1
*
9
4
*
1
4
9
5
4
0
1
7
9
*
4
2
1
*
9
Uracilpyridine 21
25
0
0
0
0
3
9
1
1
0
0
2
9
1
1
*
0
2
9
1
1
0
0
3
9
1
1
*
0
*
9
1
1
*
1
2
9
Uracilpyridine 21
50
1
2
0
0
3
9
2
2
*
0
*
9
1
1
0
0
3
9
1
2
0
0
3
9
2
2
0
1
3
9
1
2
0
1
*
9
Uracilpyridine 21
100
2
3
*
0
3
9
1
3
*
0
3
9
4
2
0
0
*
9
2
3
2
0
3
9
2
2
0
0
3
9
2
3
0
1
*
9
Uracilpyridine 22
25
0
1
*
1
*
9
0
0
0
0
1
9
0
0
0
0
*
9
1
1
0
0
1
9
0
2
0
0
2
9
1
1
1
1
1
9
Uracilpyridine 22
50
1
0
0
1
3
9
1
0
0
0
3
9
1
3
0
2
3
9
1
2
0
1
2
9
2
2
*
0
*
9
1
2
*
1
1
9
Uracilpyridine 22
100
1
2
*
0
*
9
1
3
0
0
4
9
1
1
1
0
4
9
1
2
0
1
*
9
2
2
0
1
4
9
2
3
1
0
4
9
Transgenic T3 Soybean plants harboring a PPO inhibitor tolerance trait, sprayed post in the field at the V3 leaf stage with 100 g/ha of PPO inh. 2+1% (v/v) MSO. Evaluation performed 2 and 12 Days After Treatment (DAT) and is shown as injury (%) relative to non-transgenic treated plants. Pictures taken 2 DAT.
Plant Injury (%)
Jake
Non-
AMATU_PPO2_R128A,
AMATU_PPO2_L397E,
AMATU_PPO2_L397E,
AMATU_PPO2_L397Q,
EXP
transgenic
F420I
F420V
F420M
F420M
DAT
PPO
92
10
0
5
5
2
Inh. 2
PPO
100
5
0
0
0
12
Inh. 2
The following gives a definition of the injury scores measured above:
Score
Description of injury
0
No Injury
1
Minimal injury, only a few patches of leaf injury or chlorosis.
2
Minimal injury with slightly stronger chlorosis. Overall growth points remain undamaged.
3
Slightly stronger injury on secondary leaf tissue, but primary leaf and growth points are
still undamaged.
4
Overall plant morphology is slightly different, some chlorosis and necrosis in secondary
growth points and leaf tissue. Stems are intact. Regrowth is highly probable within 1 week.
5
Overall plant morphology is clearly different, some chlorosis and necrosis on a few leaves
and growth points, but primary growth point is intact. Stem tissue is still green. Regrowth is
highly probably within 1 week.
6
Strong injury can be seen on the new leaflet growth. Plant has a high probability to
survive only through regrowth at different growth points. Most of the leaves are chlorotic/
necrotic but stem tissue is still green. May have regrowth but with noticeable injured
appearance.
7
Most of the active growth points are necrotic. There may be a single growth point that
could survive and may be partially chlorotic or green and partially necrotic. Two leaves may still
be chlorotic with some green; the rest of the plant including stem is necrotic.
8
Plant will likely die, and all growth points are necrotic. One leaf may still be chlorotic with
some green. The remainder of the plant is necrotic.
9
Plant is dead. | |
Wendell Wallach - Moral Machines: Teaching Robots Right from Wrong
Computers are already approving financial transactions, controlling electrical supplies, and driving trains. Soon, service robots will be taking care of the elderly in their homes, and military robots will have their own targeting and firing protocols. Colin Allen and Wendell Wallach argue that as robots take on more and more responsibility, they must be programmed with moral decision-making abilities, for our own safety. Taking a fast paced tour through the latest thinking about philosophical ethics and artificial intelligence, the authors argue that even if full moral agency for machines is a long way off, it is already necessary to start building a kind of functional morality, in which artificial moral agents have some basic ethical sensitivity. But the standard ethical theories don't seem adequate, and more socially engaged and engaging robots will be needed. As the authors show, the quest to build machines that are capable of telling right from wrong has begun.
| |
Asexual reproduction is a form of reproduction in which an organism creates a genetically-similar or identical copy of itself without a contribution of genetic material from another individual. It does not involve meiosis, ploidy reduction, or fertilization, and only one parent is involved genetically. A more stringent definition is agamogenesis, which refers to reproduction without the fusion of gametes.
Asexual reproduction is the primary form of reproduction for single-celled organisms such the archaea, bacteria, and protists. However, while all prokaryotes reproduce asexually (without the formation and fusion of gametes), there also exist mechanisms for lateral gene transfer, such as conjugation, transformation, and transduction, whereby genetic material is exchanged between organisms. Biological processes involving lateral gene transfer sometimes are likened to sexual reproduction (Narra and Ochman 2006). The reproductive variances in bacteria and protists also may be symbolized by + and - signs (rather than being called male and female), and referred to as "mating strains" or "reproductive types" or similar appellations.
Many plants and fungi reproduce asexually as well, and asexual reproduction has been cited in some animals, including bdelloid rotifers, which only are known to reproduce asexually, and various animals that exhibit parthenogenesis under certain conditions. In parthenogenesis, such as found in some invertebrates and vertebrates, an embryo is produced without fertilization by a male. Generally, parthenogenesis is considered a form of asexual reproduction because it does not involve fusion of gametes of opposite sexes, nor any exchange of genetic material from two different sources (Mayr 2001) however, some authorities (McGraw-Hill 2004) classify parthenogenesis as sexual reproduction on the basis that it involves gametes or does not produce an offspring genetically identical to the parent (such as a female domestic turkey producing male offspring).
A wide spectrum of mechanisms may be exhibited. For example, many plants alternate between sexual and asexual reproduction (see Alternation of generations), and the freshwater crustacean Daphnia reproduces by parthenogenesis in the spring to rapidly populate ponds, then switches to sexual reproduction as the intensity of competition and predation increases. Many protists and fungi alternate between sexual and asexual reproduction.
A lack of sexual reproduction is relatively rare among multicellular organisms, which exhibit the characteristics of being male or female. Biological explanations for this phenomenon are not completely settled. Current hypotheses suggest that, while asexual reproduction may have short term benefits when rapid population growth is important or in stable environments, sexual reproduction offers a net advantage by allowing more rapid generation of genetic diversity, allowing adaptation to changing environments.
Contents
- 1 Costs and benefits
- 2 Types of asexual reproduction
- 3 Alternation between sexual and asexual reproduction
- 4 Examples in animals
- 5 References
- 6 Credits
Costs and benefits
In asexual reproduction, an individual can reproduce without involvement with another individual, there is no fusion of gametes, and the new organism produced inherits all of its chromosomes from one parent and thus is a genetically-similar or identical copy of the parent.
Because asexual reproduction does not require the formation of gametes (often in separate individuals) and bringing them together for fertilization, nor involvement of another organism, it occurs much faster than sexual reproduction and requires less energy. Asexual lineages can increase their numbers rapidly because all members can reproduce viable offspring. In sexual populations with two genders, some of the individuals are male and cannot themselves produce offspring. This means that an asexual lineage will have roughly double the rate of population growth under ideal conditions when compared with a sexual population half composed of males. This is known as the two-fold cost of sex. Other advantages include the ability to reproduce without a partner in situations where the population density is low (such as for some desert lizards), reducing the chance of finding a mate, or during colonization of isolated habitats such as oceanic islands, where a single (female) member of the species is enough to start a population. There does not need to be energy spent in finding and courting a partner for reproduction.
A consequence of asexual reproduction, which may have both benefits and costs, is that offspring are typically genetically similar to their parent, with as broad a range as that individual receives from one parent. The lack of genetic recombination results in fewer genetic alternatives than with sexual reproduction. Many forms of asexual reproduction, for example budding or fragmentation, produce an exact replica of the parent. This genetic similarity may be beneficial if the genotype is well-suited to a stable environment, but disadvantageous if the environment is changing. For example, if a new predator or pathogen appears and a genotype is particularly defenseless against it, an asexual lineage is more likely to be completely wiped out by it. In contrast, a lineage that reproduces sexually has a higher probability of having more members survive due to the genetic recombination that produces a novel genotype in each individual. Similar arguments apply to changes in the physical environment. From an evolutionary standpoint, one could thus argue that asexual reproduction is inferior because it stifles the potential for change. However, there is also a significantly reduced chance of mutation or other complications that can result from the mixing of genes.
Conversely, Heng (2007) proposes that the resolution to the "paradox of sex" is that sexual reproduction actually reduces the drastic genetic diversity at the genome or chromosome level, resulting in the preservation of species identity, rather than the provision of evolutionary diversity for future environmental challenges. He maintains that while genetic recombination contributes to genetic diversity, it does so secondarily and within the framework of the chromosomally defined genome. That is, the asexual process generates more diverse genomes because of the less controlled reproduction systems, while sexual reproduction generates more stable genomes.
A 2004 article in the journal Nature reported that the modern arbuscular mycorrhizas fungi, which reproduces asexually, is identical to fossil records dating back to the Ordovician period, 460 million years ago (Pawlowska and Taylor 2004).
Types of asexual reproduction
Binary fission
Binary fission involves the reproduction of a living cell by division into two parts, which each have the potential to grow to the size of the original cell. Many single-celled organisms (unicellular), such as archaea, bacteria, and protists, reproduce asexually through binary fission. Exceptions are unicellular fungi such as fission yeast, unicellular algae such as Chlamydomonas, and ciliates and some other protists, which reproduce both sexually and asexually. Some single-celled organisms (unicellular) rely on one or more host organisms in order to reproduce, but most literally divide into two organisms. In addition, mitochondria and chloroplasts of eukaryote cells divide by binary fission. (See also the description under sexual reproduction.)
Budding
Some cells split via budding (for example baker's yeast), resulting in a "mother" and "daughter" cell. Budding is the formation of a new organism by the protrusion of part of another organism, with the protrusion normally staying attached to the primary organism for a while, before becoming free. The new organism is naturally genetically identical to the primary one (a clone) and initially is smaller than the parent. Budding is also known on a multicellular level; an animal example is the hydra, which reproduces by budding. The buds grow into fully matured individuals, which eventually break away from the parent organism.
Vegetative reproduction
Vegetative reproduction is a type of asexual reproduction found in plants where new independent individuals are formed without the production of seeds or spores. Examples for vegetative reproduction include the formation of plantlets on specialized leaves (for example in kalanchoe), the growth of new plants out of rhizomes or stolons (for example in strawberry), or the formation of new bulbs (for example in tulips). The resulting plants form a clonal colony.
Spore formation
Many multicellular organisms form spores during their biological life cycle in a process called sporogenesis. Exceptions are animals and some protists, which undergo gametic meiosis immediately followed by fertilization. Plants and many algae on the other hand undergo sporic meiosis, where meiosis leads to the formation of haploid spores rather than gametes. These spores grow into multicellular individuals (called gametophytes in the case of plants) without a fertilization event. These haploid individuals give rise to gametes through mitosis. Meiosis and gamete formation therefore occur in separate generations or "phases" of the life cycle, referred to as alternation of generations. Since sexual reproduction is often more narrowly defined as the fusion of gametes (fertilization), spore formation in plant sporophytes and algae might be considered a form of asexual reproduction (agamogenesis) despite being the result of meiosis and undergoing a reduction in ploidy. However, both events (spore formation and fertilization) are necessary to complete sexual reproduction in the plant life cycle.
Fungi and some algae can also utilize true asexual spore formation, which involves mitosis giving rise to reproductive cells called mitospores that develop into a new organism after dispersal. This method of reproduction is found, for example, in conidial fungi and the red alga Polysiphonia, and involves sporogenesis without meiosis. Thus, the chromosome number of the spore cell is the same as that of the parent producing the spores. However, mitotic sporogenesis is an exception and most spores, such as those of plants, most Basidiomycota, and many algae, are produced by meiosis.
Fragmentation
Fragmentation is a form of asexual reproduction where a new organism grows from a fragment of the parent. Each fragment develops into a mature, fully grown individual. Fragmentation is seen in many organisms, such as animals (some annelid worms and starfish), fungi, and plants. Some plants have specialized structures for reproduction via fragmentation, such as gemmae in liverworts. Most lichens, which are a symbiotic union of a fungus and photosynthetic algae or bacteria, reproduce through fragmentation to ensure that new individuals contain both symbionts. These fragments can take the form of soredia, dust-like particles consisting of fungal hyphae wrapped around photobiont cells.
Parthenogenesis
Parthenogenesis is a form of agamogenesis in which an unfertilized egg develops into a new individual. Parthenogenesis occurs naturally in many plants, invertebrates (e.g. water fleas, aphids, stick insects, some ants, bees and parasitic wasps), and vertebrates (e.g. some reptiles, amphibians, fish, very rarely birds). In plants, apomixis may or may not involve parthenogenesis.
Parthenogenesis is one example of agamogenesis, the term for any form of reproduction that does not involve a male gamete. An example is apomixis.
Apomixis and nucellar embryony
Apomixis in plants is the formation of a new sporophyte without fertilization. It is important in ferns and in flowering plants, but is very rare in other seed plants. In flowering plants, the term "apomixis" is now most often used for agamospermy, the formation of seeds without fertilization, but was once used to include vegetative reproduction. An example of an apomictic plant would be the triploid European dandelion.
Apomixis mainly occurs in two forms. In gametophytic apomixis, the embryo arises from an unfertilized egg within a diploid embryo sac that was formed without completing meiosis. In nucellar embryony, the embryo is formed from the diploid nucellus tissue surrounding the embryo sac. Nucellar embryony occurs in some citrus seeds. Male apomixis can occur in rare cases, such as the Saharan cypress where the genetic material of the embryo are derived entirely from pollen.
The term "apomixis" is also used for asexual reproduction in some animals, notably water-fleas, Daphnia.
Alternation between sexual and asexual reproduction
Some species alternate between the sexual and asexual strategies, an ability known as heterogamy, depending on conditions. For example, the freshwater crustacean Daphnia reproduces by parthenogenesis in the spring to rapidly populate ponds, then switches to sexual reproduction as the intensity of competition and predation increases.
Many protists and fungi alternate between sexual and asexual reproduction. For example, the slime mold Dictyostelium undergoes binary fission as single-celled amoebae under favorable conditions. However, when conditions turn unfavorable, the cells aggregate and switch to sexual reproduction leading to the formation of spores. The hyphae of the common mold (Rhizopus) are capable of producing both mitotic as well as meiotic spores.
Many algae similarly switch between sexual and asexual reproduction. Asexual reproduction is far less complicated than sexual reproduction. In sexual reproduction one must find a mate.
Examples in animals
A number of invertebrates and some less advanced vertebrates are known to alternate between sexual and asexual reproduction, or be exclusively asexual. Alternation is observed in a few types of insects, such as aphids (which will, under favorable conditions, produce eggs that have not gone through meiosis, essentially cloning themselves) and the cape bee Apis mellifera capensis (which can reproduce asexually through a process called thelytoky). A few species of amphibians and reptiles have the same ability (see parthenogenesis for concrete examples). A very unusual case among more advanced vertebrates is the female turkey's ability to produce fertile eggs in the absence of a male. The eggs result in often sickly, and nearly always male turkeys. This behavior can interfere with the incubation of eggs in turkey farming (Savage 2008).
There are examples of parthenogenesis in the hammerhead shark (Eilperin 2007) and the blacktip shark (Chapman et al. 2008). In both cases, the sharks had reached sexual maturity in captivity in the absence of males, and in both cases the offspring were shown to be genetically identical to the mothers.
Bdelloid rotifers reproduce exclusively asexually, and all individuals in the class Bdelloidea are females. Asexuality arose in these animals millions of years ago and has persisted since. There is evidence to suggest that asexual reproduction has allowed the animals to develop new proteins through the Meselson effect that have allowed them to survive better in periods of dehydration (Pouchkina-Stantcheva et al. 2007).
References
- Chapman, D. D., B. Firchau, and M. S. Shivji. 2008. Parthenogenesis in a large-bodied requiem shark, the blacktip Carcharhinus limbatus. Journal of Fish Biology 73(6): 1473. See report in Science Daily: "Virgin birth" By shark confirmed: Second case ever. Retrieved January 15, 2009.
- Eilperin, J. 2007. Female sharks can reproduce alone, researchers find. Washington Post May 23, 2007, p. A02. Retrieved January 16, 2008.
- Graham, L., J. Graham, and L. Wilcox. 2003. Plant Biology. Upper Saddle River, NJ: Pearson Education. ISBN 0130303712.
- Heng, H. H. 2007. Elimination of altered karyotypes by sexual reproduction preserves species identity. Genome 50: 517-524.
- Mayr, E. 2001. What Evolution Is. New York: Basic Books. ISBN 0465044255.
- McGraw-Hill (Publisher). 2004. McGraw-Hill Concise Encyclopedia of Science and Technology, 5th Edition. McGraw Hill Professionals. ISBN 0071429573.
- Narra, H. P., and H. Ochman. 2006. Of what use is sex to bacteria? Current Biology 16: R705–710. PMID 16950097.
- Pawlowska, T., and J. Taylor. 2004. Organization of genetic variation in individuals of arbuscular mycorrhizal fungi. Nature 427(6976): 733-737.
- Pouchkina-Stantcheva, N. N., B. M. McGee, C. Boschetti, et al. 2007. Functional divergence of former alleles in an ancient asexual invertebrate. Science 318: 268-271. Retrieved January 15, 2009.
- Raven, P. H., R. F. Evert, and S. E. Eichhorn. 2005. Biology of Plants, 7th edition. New York: W.H. Freeman and Company. ISBN 0716710072.
- Savage, T. F. 2008. [http://oregonstate.edu/instruct/ans-tparth/index.html A guide to the recognition of parthenogenesis in incubated turkey eggs. Oregon State Universtiy. Retrieved January 16, 2009.
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In recent years, a global emphasis on combatting climate change has led to an elevated focus on more sustainable processes among the commercial maritime shipping industry. At a macro level, industry collaborations like the Sea Cargo Charter have committed to embracing more sustainable practices. Meanwhile, at a micro level, individual organisations are challenged to find new ways to think and act sustainably in their day-to-day workflows.
For many, however, the process of achieving true sustainability in the maritime industry seems a bit overwhelming, full of complexity and unknowns. However, it doesn’t have to be this way. The industry, and the world at large, is changing, and you can be a part of that movement by making changes at the micro level which ripple into substantial positive impact.
But with so many diverse sustainability-driven initiatives and technologies cropping up, where do you start? What should you focus on? It’s important to remember that a maritime organisation’s sustainability strategy is intrinsically linked to its digitalisation strategy, and that these initiatives are a journey not a destination.
In this infographic we highlight four major breakthroughs that will shape the maritime industry by 2030.
To prepare for future regulations and support more sustainable workflows, maritime shipping organisations benefit from access to high quality data that enables them to evaluate the environmental impact of each and every decision—from pre-fixture through ongoing voyage operations.
In collaborating with hundreds of maritime organizations across the industry on the digital technologies that drive their business forward, we have gathered a list of actionable steps that you can take right now to make progress towards a sustainable future.
1.) Build a foundation for long-term tracking and data management.
It’s exciting to imagine the transformative possibilities the next decade could bring. However, none of these can be achieved without the proper digital foundation. In a data-driven world, any initiative designed to make an impact must be measured, tracked, and reported on. A digital system that can streamline this data collection and management can be the foundation that positions your sustainability efforts for success.
2.) Consider your sustainability goals in context of your business.
Because effective change management cannot come at the expense of business operations, no two paths to organizational sustainability are exactly alike. It’s imperative that you begin your journey with a thorough self-examination. What matters to your organisation? What will enable you to succeed? What are your short-term and long-term goals? Answering these questions helps to identify investments that will deliver immediate value and long-term dividends, and most importantly, they will help you find your first step towards lasting change.
3.) Recognise that sustainability isn’t an isolated initiative.
Instead, it is becoming pervasive and integral to the way business is conducted in the modern world. By expanding your view of sustainability beyond a series of one-time initiatives, you’ll be able to view each of your core functional areas in a new light, bring manageable change to daily decisions, and capitalize on its full potential by incorporating sustainability data into your centralised, digital ecosystem.
4.) Crowdsource your future, internally.
The individuals that understand your needs and requirements are right inside your business. Cross-functional and cross-managerial engagement are integral to every phase of your digital journey—from surfacing needs in the planning phase to promoting utilisation through continuous education, and soliciting feedback to gauge long-term success.
Looking at the volume of work to be done in order to achieve long-term, industry-wide sustainability goals can be intimidating, to say the least. While it is true that breakthroughs like low-emission ships and alternative fuels are time and resource-intensive, there are many ways that maritime shipping organisations can make a measurable impact today.
Don’t let the size of long-term sustainability goals prevent you from taking immediate steps to better understand your emissions and make decisions to reduce your environmental impact; begin your journey by taking manageable steps in line with your existing process.
We at Veson Nautical are ready for a future focused on sustainability. Read our whitepaper to learn more about the next decade’s most exciting advancements and how—with a few small steps—we can make great strides, together. | https://www.tradewindsnews.com/sponsor-content/4-ways-maritime-organisations-can-drive-sustainability/2-1-974971 |
“Your work is so Dada, its just weird…” Even though the sentence was uttered playfully and with no foul intentions, it hit me. It sounded dismissive; in my ears, my friend just admitted disinterest. Calling something “weird” suggests withdrawal. The adjective forecloses a sense of urgency and classifies the work as a shallow event: the work is funny and quirky, slightly odd and soon becomes background noise, ’nuff said. I tried to ignore the one word review, but I will never forget when it was said, or where we were standing. I wish I had responded: “I think we already know too much to make art that is weird.” But unfortunately, I kept quiet.
In his book Noise, Water, Meat (1999), Douglas Kahn writes: “We already know too much for noise to exist.” A good 15 years after Kahn’s writing, we have entered a time dominated by the noise of crises. Hackers, disease, trade stock crashes and brutalist oligarchs make sure there is not a quiet day to be had. Even our geological time is the subject to dispute. But while insecurity dictates, no-one would dare to refer to this time as the heyday of noise. We know there is more at stake than just noise.
This state is reflected in critical art movements: a current generation of radical digital artists is not interested in work that is uninformed by urgency, nor can they afford to create work that is just #weird, or noisy. The work of these artists has departed from the weird and exists in an exchange that is, rather, strange. it invites the viewer to approach with inquisitiveness – it invokes a state of mind: to wonder. Consequently, these works break with tradition and create space for alternative forms, language, organisation and discourse. It is not straightforward: it is the art of creative problem creation(Jon Satrom during GLI.TC/H).
In 2016 it is easy to look at the weird aesthetics of Dada; its eclectic output is no longer unique. The techniques behind these gibberish concoctions have had a hundred years to become cultivated, even familiar. Radical art and punk alike have adopted the techniques of collage and chance and applied them as styles that are no longer inherently progressive or new. As a filter subsumed by time and fashion, Dada-esque forms of art have been morphed into weird commodities that invoke a feel of stale familiarity.
But when I take a closer look at an original Dadaist work, I enter the mind of a stranger. There is structure that looks like language, but it is not my language. It slips in and out of recognition and maybe, if I would have the chance to dialogue or question, it could become more familiar. Maybe I could even understand it. Spending more time with a piece makes it possible to break it down, to recognize its particulates and particularities, but the whole still balances a threshold of meaning and nonsense. I will never fully understand a work of Dada. The work stays a stranger, a riddle from another time, a question without an answer. The historical circumstances that drove the Dadaists to create the work, with a sentiment or mindset that bordered on madness, seems impossible to translate from one period to the next. The urgency that the Dadaists felt, while driven by their historical circumstances, is no longer accessible to me. The meaningful context of these works is left behind in another time. Which makes me question: why are so many works of contemporary digital artists still described—even dismissed—as Dada-esque? Is it even possible to be like Dada in 2016?
The answer to this question is at least twofold: it is not just the artist, but also the audience who can be responsible for claiming that an artwork is a #weird, Dada-esque anachronism. Digital art can turn Dada-esque by invoking Dadaist techniques such as collage during its production. But the work can also turn Dada-esque during its reception, when the viewer decides to describe the work as “weird like Dada.” Consequently, whether or not today a work can be weird like Dada is maybe not that interesting; the answer finally lies within the eye of the beholder. It is maybe a more interesting question to ask what makes the work of art strange? How can contemporary art invoke a mindset of wonder and the power of the critical question in a time in which noise rules and is understood to be too complex to analyse or break down?
The Dadaists invoked this power by using some kind of ellipsis (…): a tactic of strange that involves the withholding of the rules of that tactic. They employed a logic to their art that they did not share with their audience; a logic that has later been described as the logic of the madmen. Today, in a time where our daily reality has changed and our systems have grown more complex, the ellipses of mad logic (dysfunctionality) are commonplace. Weird collage is no longer strange; it is easily understood as a familiar aesthetic technique. Radical Art needs a provocative element, an element of strange that lures the viewer in and makes them think critically; that makes them question again. The art of wonder can no longer lie solely in ellipsis and the ellipsis can no longer be THE art.
This is particularly important for digital art. During the past decades, digital art has matured beyond the Dadaesque mission to create new techniques for quaint collage. Digital artists have slowly established a tradition that inquisitively opens up the more and more hermetically closed—or black boxed—technologies. Groups and movements like Critical Art Ensemble (1987), Tactical Media (1996), Glitch Art (since ±2001, and later a subgenre that is sometimes referred to as Tactical Glitch Art) and #Additivism (2015) (to name just a few) work in a reactionary, critical fashion against the status quo, engaging with the protocols that facilitate and control the fields of, for instance, infrastructure, standardization, or digital economies. The research of these artists takes place within a liminal space, where it pivots between the thresholds of digital language, such as code and algorithms, the frameworks to which data and computation adhere and the languages spoken by humans. Sometimes they use tactics that are similar to the Dadaist ellipsis. As a result, their output can border on Asemic. This practice comes close to the strangeness that was an inherent component of an original power of Dadaist art.
But an artist who still insists on explaining why a work is weirdly styled like Dada is missing out on the strange mindset that formed the inherently progressive element of Dada. Of course a work of art can be strange by other means than the tactics and techniques used in Dada. Dada is not the father of all progressive work. And not all digital art needs to be strange. But strange is a powerful affect from which to depart in a time that is desperate to ask new critical questions to counter the noise.
Thanks to Amy J. Elias and Jonathan P. Eburne
NotesImages in this article are part of the exhibition Filtering Failure curated by Julian van Aalderen and Rosa Menkman in 2011.
Catalogue of exhibition here: | https://www.furtherfield.org/what-is-a-question-a-stranger-like-dada-weird-like-quaint-collage-%C2%AF_-%CD%A1%E0%B0%A0%CA%98%E1%B4%A5-%E2%8A%99%E0%B2%A5%E2%80%B6%CA%94%EF%BE%89%CC%B5%CD%87%CC%BF%CC%BF%CC%BF-%CC%BF-%CC%BF/ |
Once the decision has been made to create a digital image collection, its scope and form need to be tailored to the particular institution: the more time spent in review and analysis before embarking on the first scan, the more successful a project is likely to be. Remember that projects may be carried out in partnership or collaboration with other institutions or initiatives, and that this may allow sharing of both costs and expertise.
Collection Selection
The first step is to select the collection, collections, or part of a collection to be digitized. Consider the level of interest in the selection and its relevance to the scanning institution's mission. Make sure that the scale of the proposed project is practical, considering the broader technical environment: the operating systems, networks, and bandwidth in place, and overall budgets and priorities. It is advisable to think through an overall strategy but to begin with smaller projects and work up gradually to a more ambitious program.
Conservation and Access Status
Collections that are already in good condition and have consistent metadata control make for far less arduous imaging projects. Ensure that the items are not too physically fragile to withstand the imaging process without damage, and decide which scanning method is most appropriate (see
Selecting Scanners
). Appraise the collection's organization: well-organized collections facilitate a robust linking of physical object and digital surrogate through such strategies as consistent file-naming protocols, while chaotic collections do not. Maintaining such relationships between analog and digital assets is crucial for managing hybrid collections. Completing conservation and cataloguing of any selected collection before beginning the scanning process is highly recommended.
Legal Status
Discover whether any legal clearances are required to reproduce the originals or to modify and display the reproductions. Be aware that many license agreements are of limited duration, which may be a problem if the intention is that a digital image collection be available indefinitely. Projects are much more straightforward if clearance requirements are minimal, as for instance when the items to be digitized are in the public domain or the scanning institution owns reproduction rights.
Project Team and Workflow
Identify the team that will be required to complete the project. Digitizing projects generally require the expertise of many different departments and/or individuals, whose availability should be considered when plotting out the project timeline and workflow. Decide which, if any, of the many tasks involvedconservation, photography, scanning, cataloguing, metadata capture, storageare to be outsourced. It will be necessary to review workflow constantly in order to recognize and resolve weaknesses and bottlenecks.
Standards Selection
It will be necessary to decide which imaging (file format, resolution, naming protocols, and so on) and metadata standards to employ, taking into account the nature of the original material, the staff time available for indexing and cataloguing, and the likely users of the collection. (See
Image Capture and Selecting a Metadata Schema
.) Certain standards may already be in place within an institution, and participating in certain partnerships or collaborations may prompt the selection of standards already in use within the larger group.
Digital Asset Management
The standards in use within the larger group may also influence the selection of hardware, software, and, perhaps most critically, an image or digital asset management (
DAM
) system. It is important to remember that DAM software cannot develop asset management strategies (though it can be used to implement or enforce them) and that whatever management system is used, its usefulness will depend on the quality of metadata it contains.
DAM systems can track digital image creation and modification, record the location of master and derivative files, allow search and retrieval of files, monitor migration schedules, control access, and so forth. Turnkey or customizable off-the-shelf DAM systems are available at a broad range of prices and levels of complexity, and it is also possible to utilize desktop database software or more powerful client/server systems to create in-house customized solutions, or to employ some combination of commercial and in-house systems. XML-based solutions such as native-XML or
XML-enabled
databases are likely to become more popular in the future.
The most appropriate image management solution will be dictated by the available budget, the scale of the project and its projected growth, the available technical infrastructure and support, the projected demand, and similar issues. Most institutions will want to incorporate their DAM system into a general, institution-wide automation or digital library plan. This will require some level of integration with existing or planned collection and library management systems, online public access catalogues (
OPAC
), publishing systems, and perhaps business or administrative systems. The use of consistent data structure and content standards ensures flexibility by facilitating the exchange and migration of data and thus promoting interoperability and
resource sharing
within (and between) institutions.
User Requirements
All aspects of a digital imaging project will need to take into consideration the needs of each class of potential user, but these will most particularly guide decisions about presentation and delivery. Understanding user needs requires probing the assumptions of differing groups, which may be achieved through user studies. These may reveal particular requirements or limitations, such as the desired level of image quality, necessary information-searching facilities, or a predefined network infrastructure. For example, medium-resolution images of a particular collection may be sufficient for classroom use by undergraduate students, but they may contain too little information for a conservator exploring the technical construction of a work. Security protocols can be used to give different image and metadata access to the various users of a collection, if this is deemed necessary (see
Security Policies and Procedures
). It will be necessary to select which data elements should display to the various user groups, which should be searchable, and what kinds of searches should be possible.
Other requirements or preferences may also be revealed through user studies. Will users want to integrate image display with other institutional information? For example, would users want to display a record from a curatorial research database or library management system alongside the image? Will users wish to be able to integrate search results into word processing or other documents, and, therefore, should copying or downloading of the image and record be facilitated (which might have legal implications)? Do users require thumbnail images for browsing, and, if so, what type of identification should accompany each image? Would image processing or image manipulation functions (such as changing colors, zooming, or
annotation
) be helpful to users? It may not be desirable or even possible to fulfill all such desires, but it is useful to be aware of them. (See
Delivery
.)
Digital Preservation
It will be absolutely necessary to develop a strategy for ensuring long-term access to, and preservation of, assets. This will require choosing that combination of tacticssuch as documentation,
redundant storage
,
refreshing
, migration, | https://www.getty.edu/research/publications/electronic_publications/introimages/planning.html |
Are we ready to have this confrontational conversation about our relationship with fashion? Fact is, we all have a relationship with fashion. It’s true even for those of us who couldn’t care less about what we drape on our bodies. In one way or another, we all consume and support the fashion industry and this particular industry is one of the most polluting in the world. The fashion industry has a long way to go before it can rectify the damaged it has inflicted. But all gory and sad endings aside, not all is lost. A spark of hope emerges from the gloom and doom. The next generation of aspiring designers – conscious and aware of their social and ethical responsibility to the environment are paving a new way forward.
How did you get into sustainable fashion?
Joshua: I chose to work on sustainable fashion because I feel that sustainable fashion is the future. It’s not a choice anymore if I want to stay in the fashion industry. It’s just the way forward.
Ruby: To be honest, I agree we need to be sustainable now but why I started was because I wanted to save cost. I use dead-stock material and fabric to up-cycle them.
Sarah: In our second year of university, we had to pick a track for our project. I chose fashion ecology which led onto sustainable fashion. I specifically chose an alternative material to work with because I’m interested to see how we can push that further.
What does sustainable fashion mean to you?
Joshua: There are many aspects to sustainable fashion, but my main concern is being environmentally friendly and responsible when designing and producing.
Ruby: Same, it’s the environment. To produce less waste and be mindful of the environment when producing garments.
Sarah: For me, it’s about creating change. The fashion industry has created has a lot of environmental and ethical issues, but designers have the capacity to address these issues.
Describe your individual projects to us.
Joshua: My method is called “spaghetti cutting”. It is a method to use the entire width of the fabric. I came up with a formula and I determined the perfect width to cut the strips of fabric. I found that 7 inches is the best width length and that reduces the amount of waste significantly. The wastage was only from the collar, neck, and armhole which is very little compared to the conventional method of pattern cutting. In a real-life situation, the conventional way will not fully utilize the fabric but the “spaghetti cutting” method will, no matter what the outcome of the garment is.
Sarah: What I did for my project was to explore agar, alginate, and starch-based bioplastics. I explored what was accessible to me at the time. Through the process, I was able to formulate and create many different recipes. I was also able to categorize them in their own significant properties or design. Through that, I was able to apply that to fashion design. I created small embroidery pieces and got them laser cut and had them directly on fabrics. The idea was to rethink and repurpose the garments by taking out the embroidery pieces. I also thought about using starch to make buttons and fasteners. I 3-D printed and created my own silicon mold of buttons and then using tapioca starch to make them so that the garment would be 100% disposable. It’s a different possibility on how we can wear our clothes. Lastly, I also found an alternative material for leather. It had the same durability and texture as leather. Through this process, it has created new challenges for me because these garments are not meant to last or to be repaired. They have some level of durability, but it is disposable at any point of time. To create colors, I used pigment powder. Next for me would be to explore different possibilities on how to dispose of the garments besides just in soil and worm compost.
Ruby: For my project, I learned that people threw away their clothes because they had holes in them. So, I tried to reinvent the holes. I make the holes look “perfect” by adding new elements to the old garment. I thought this was a better way because apart from the hole, the rest of the garment still was in good condition. So, I reinvent the garment by adding reflective threads, embellishments, and embroidery. I tried to take the problem and turning it into a design as opposed to an accident.
Human consumption vs designing process – should we change our consumption, or must designers change their designs?
Joshua: I think both must change. Human consumption is a problem. It’s like supply and demand. If more people want, brands will produce more. But my focus is to reduce waste even in doing fast fashion. Most fashion production has a 25% wastage and that is not inclusive of over-production for example if people don’t buy. My aim is to reduce waste in general. It depends on the design––spaghetti cutting allows me to reduce wastage to only 10%. I think brands have the power to change human mindsets. Fashion wasn’t always so affordable. Fast fashion produces huge quantities at such a low cost which makes it possible for them to sell clothes so cheaply. So, they allow people to buy more because it’s so affordable.
Ruby: I agree with everything that’s been said. I noticed people throw out most of their clothes because of one small hole or rip. I thought to design and up-cycle clothes so they can become something new. I used deadstock fabrics from fashion companies. Essentially I used fabric that was already made instead of producing new material.
Amelia: So it’s a team effort. Designers, consumers, and everyone in between have to play a part in wanting to consciously be sustainable.
Sarah: I was targeting textile waste. Human consumption is ever-present, and it’s accelerated by fast fashion. So, I thought about rethinking the material of the clothes we wear and how long they last. Since people are not keeping their garments for long, I made small batches of clothes made from agar and bioplastics, so the garments are biodegradable. I experimented by placing the clothes in worm compost and it would take about 2 weeks to a month to decompose without any added additives.
What is fashion’s biggest challenge with sustainability?
Joshua: Finding the balance between being sustainable and making profits. Hence why I feel some brands do greenwashing.
Ruby: Being sustainable is more expensive. The way we execute sustainability could also be quite costly. I think cost would be the main problem.
Sarah: I think it’s not easy to be accountable when we try to be sustainable. I think it’s hard to be forward-looking. So, how one sets new goals after completing a specific sustainable goal, to continuously be sustainable. I think that can be quite a laborious and tiring process.
Are we educated enough about sustainability?
Joshua: I think people are educated but not everyone cares. I think people in Southeast Asia know about sustainable fashion. We have exposure to sustainability. However, people are so worried about the cost of a garment. Not how much it costs the environment. The price of the garment comes first. But knowing and wanting to do your part is different. The mentality is “why should I care if everyone is doing it? Me alone, isn’t going to make a difference?” I think that is a hard mindset to change.
Ruby: I think they don’t know in-depth about what is happening. Maybe they know about what’s happening on the surface. For example, they don’t know how much water is used to make cotton. They don’t know in-depth facts on how it’s affecting the earth.
Sarah: We’re still stuck in the cycle of consuming. Also, nowadays, we buy a lot of clothes and then we would sell them all at once. There is a rise in secondhand culture, trading, and re-selling our clothes. While this seems to be good, I think it can also be used as an escape. This can be used as a way for us to brainwash ourselves thinking that we are sustainable but it’s just an escape so that we can buy more clothes. Maybe we need more education on consumption, so we don’t get stuck in this vicious cycle of buying and selling.
Joshua: I would like to add, maybe a lot of brands try to sell that they are sustainable, but we will never know if they really are. I think brands know the importance of sustainability but at the end of the day, it’s a business. Perhaps, I am optimistic now as I am young and fresh out of university, so the sustainability aspect is my priority but who knows if I get into the business, it might be harder to achieve. I have an inner conflict about whether it would be hard to have a business built on sustainable grounds.
Where do you envision yourself and fashion in five years time?
Ruby: I don’t think there’s going to be much of a difference in the fashion industry. Not a drastic change, at least. Perhaps about the same. I think it’s hard for people to adapt to slow fashion. Where I would be is to educate people on how to mend their clothes. Workshops to make new garments out of their existing clothes and crafting. I envision myself doing that.
Sarah: Hopefully I would be more involved in a community where I can have a network to increase my resources. Especially while doing this project, I had to reach out to a lot of people to get resources to see better results. There’s a strong focus on new technology for fashion. I hope I’d be more involved to learn about new technologies to facilitate alternative materials.
Joshua: I don’t see myself as a fashion designer. I would like to be a pattern cutter. I am so heavily invested in pattern cutting because I think how you cut the piece of the pattern is an integral part of where most waste is produced. Hence why, I hope to stay in the production line. I would like to incorporate better strategies to reduce waste. I would want to work with fast fashion brands because they produce the most waste, so I’d like to innovate more methods to reduce waste within the fast fashion industry. | https://mulazine.com/fashion-and-beauty/fashion-fades-style-is-eternal-but-sustainability-is-the-future/ |
Post-traumatic Stress Disorder Awareness Month
The National Institute of Mental Health (NIMH) defines PTSD as a disorder “that develops in some people who have experienced a shocking, scary, or dangerous event.” While feeling stress and anxiety is normal in these situations, those with PTSD continue to experience extreme stress and fear even after the event has passed and the individual is no longer in danger.
What Are the Symptoms of PTSD?
The Anxiety and Depression Association of America (ADAA) states that PTSD is different than other types of stress and anxiety, both in its longevity and in how it manifests.
A PTSD diagnosis includes symptoms that are present for at least one month after the event has passed. These symptoms may not occur right away, and some people experience the onset of PTSD weeks, months, or even years after the traumatic event itself. There are three main symptoms associated with PTSD:
- Re-experiencing the trauma—One of the most common symptoms of PTSD; this occurs through unwanted, troubling recollections, nightmares, and/or flashbacks.
- Avoidance of things that remind the individual of the trauma—Sometimes referred to as emotional numbness, this symptom is the intentional withdrawal from people, places, and activities that may remind the person of the traumatic event.
- Increased arousal—Those with PTSD may have difficulty sleeping, trouble with concentration and focus, feel constantly on edge, and be easily irritated and angered.
Who Is Affected by PTSD?
Anyone who experiences or witnesses a traumatic event is at risk for PTSD. Numerous experiences can cause PTSD and some of the most common include combat, terror attack, natural disasters, abuse, assault, and sudden loss.
According to PTSD Alliance around 5% of men and 10% of women will develop PTSD in the United States. That means 6-7 million Americans have PTSD, with roughly eight percent suffering from the disorder at any given time.
- Children: The National Center for PTSD estimates that 15% to 43% percent of girls and 14% to 43% percent of boys experience at least one traumatic event during their childhood and adolescent years. Of these children, 3% to 15% percent of the girls and 1% to 6% percent of the boys develop PTSD. The disorder appears correlated with the severity of the event, the parents’ reaction, and how close the child was to the trauma.
- Women: Women, overall, experience different traumatic events than men. Five out of 10 women experience a traumatic event.
- Men: Men are less likely to develop symptoms of PTSD and four percent of men who experience trauma are diagnosed with the disorder.
- Military: When it comes to military personnel, the risk of developing PTSD increases. According to Veteran’s Affairs (VA), 10% to 18% of those deployed in Afghanistan and Iraq were diagnosed with PTSD in a given year.
Where to Find Help for PTSD?
When it comes to finding help for PTSD, the most important thing to remember is that it is treatable. The course of treatment for two people may be very different. What works for one person may not work for another and it may take a few different approaches before finding the right one.
The VA offers therapy for PTSD: Cognitive Behavior Therapy (CBT) is considered the most effective treatment and involves learning how thoughts about a traumatic event cause certain reactions to occur, worsening PTSD symptoms. By replacing those specific thoughts, the reactions occur less, and the symptoms dissipate. Other treatments, such as eye movement desensitization and reprocessing (EMDR) and certain medications may also be recommended.
Continue Spreading Awareness
The United States Senate has officially designated the month of June as PTSD Awareness Month. It’s a month dedicated to raise public awareness of PTSD and the treatments that are available. By focusing on education, outreach, and spreading the word about PTSD throughout the month, supporters can help others overcome this severe, yet treatable, disorder.
Information on OnlineCounselingPrograms.com is not intended to be a substitute for professional medical advice. Always consult your physician or other qualified professionals with any questions you may have regarding a Post-traumatic Stress disorder. | https://onlinecounselingprograms.com/resources/posttraumatic-stress-disorder-awareness-month/ |
Why now?
Decarbonisation and collaboration in wind and aviation
To further decarbonise, a collaborative approach between wind and aviation is crucial. Following two successful years of workshop- style discussions, RenewableUK’s Wind and Aviation event is evovling to a webinars series where stakeholders and industry experts discuss potential challenges and suggest practical solutions on a range of crucial topics including helicopter operations, drones & UAS, lighting & marking and surveillance & radar.
Dates and topics
- Thursday 29 April | Helicopter operations: Wind as a customer of aviation services
- Thursday 19 May | Unmanned aerial systems
- Wednesday 16 June | Lighting & Marking
- Thursday 9 September | SurveIllance & Radar
Why attend?
RenewableUK’s new Wind & Aviation webinar series is the only dedicated event of its kind offering:
- Four, short, focussed webinars homing in on business-critical discussions
- Access to aviation stakeholders and industry experts in exclusive sessions and interactive Q&As
- Access to exclusive business intelligence and cutting-edge tech insights
- Free to all RenewableUK members
- Supported and designed by the industry for the industry.
Who will you meet?
Key aviation stakeholders, airports, aircraft, radar, drone and UAS manufacturers and operators, project developers, government, consultancies, national and regional business clusters, training providers, health & safety experts, professional associations, and trade press. | https://events.renewableuk.com/aviation21 |
On September 3, the Federal Motor Carrier Safety Administration (FMCSA) proposed a pilot program providing temporary relief from the 14-hour driving window applicable to drivers of commercial motor vehicles (CMVs), and on September 10, FMCSA proposed a second pilot program to allow drivers aged between 18 and 20 to operate CMVs in intrastate commerce.
June 2020
Beyond NAFTA: USMCA Goes into Effect July 1
Key Notes:
- Manufacturers across North America must reanalyze their products to determine whether they continue to qualify for duty-free treatment under USMCA.
- New USMCA certificates of origin are required to replace NAFTA certificates of origin.
- Many companies must examine their supply chains to confirm continuing eligibility for duty-free treatment.
On July 1, the United States-Mexico-Canada Agreement (USMCA) goes into effect. It will have a significant impact on manufacturers across North America as they reanalyze their products to determine whether they qualify for duty-free treatment.
May 2020
FMCSA Amends Its Driver Hours of Service Rules
On May 14, 2020, the Federal Motor Carrier Safety Administration (FMCSA) issued a final rule revising the hours of service (HOS) regulations for commercial motor vehicle drivers that is intended “to provide greater flexibility for drivers subject to those rules without adversely affecting safety.” We provide a summary of the changes to the HOS regulations, which include expanding the short-haul exception and the driving window during adverse driving conditions, requiring a 30-minute break after eight hours of driving time (instead of on-duty time) and modifying the sleeper berth exception.
April 2020
Government Measures Worldwide in Response to COVID-19
As countries respond to the COVID-19 pandemic, business operations must adapt to changing rules and new restrictions. We are teaming with a group of international law firms to provide and regularly update a chart compiling key measures implemented by countries around the world.
Our chart addresses:
- Economic measures, including loan facilities, tax deferrals, stimulus offerings and similar actions.
- Labor and employment and employee benefit measures.
- Health and safety measures, including travel restrictions, business and other closures, and essential business categories.
- Export and import developments, including new restrictions on the export of personal protective equipment (PPE) and medicine and reductions on import duties.
The summary is not intended as legal advice but as a practical resource for companies adapting to the global pandemic.
FMC to Address COVID-19 Port Congestion by Engaging Industry Stakeholders
On March 31, 2020, the Federal Maritime Commission (FMC) issued an order announcing that it will engage supply chain stakeholders to address disruptions that the COVID-19 pandemic may have on cargo movement through U.S. ports. The order authorizes Commissioner Dye to hold discussions with supply chain stakeholders to identify operational solutions to cargo delivery system challenges and to form one or more FMC supply chain innovation teams to support the efforts.
March 2020
FMCSA Temporarily Extends Commercial Drivers’ Licenses, Permits and Medical Certifications
On March 24, 2020, the Federal Motor Carrier Safety Administration (FMCSA) granted a temporary waiver that effectively extends the validity of commercial driver’s licenses (CDLs), commercial learner’s permits (CLPs) and medical certificates for drivers of commercial motor vehicles (CMVs). The waiver will expire on June 30, 2020.
CARES Act: Implications for Businesses
The Coronavirus Aid, Relief, and Economic Security Act (CARES Act), a roughly $2 trillion coronavirus response bill signed into law yesterday, is intended to provide widespread emergency relief for Americans and the country’s economy. In addition to its benefits for individuals, the bill provides aid for small businesses, large corporations, hospitals and public health agencies, and state and local governments. Among its provisions for small businesses are emergency grants and a forgivable loan program for companies with 500 or fewer employees, along with revised rules for expenses and deductions designed to help companies keep employees on the payroll and remain open during the crisis. Large corporations stand to benefit from approximately $500 billion in loans and other funding repayable to the government and subject to public disclosures and other requirements. Community and private health systems will receive help to enable them to better respond to the increasing demands they face. Provisions for state and local governments include direct aid to bolster cash reserves depleted by COVID-19 response efforts, along with Community Development Block Grants, school funding and money for programs for children and families. In this bulletin, we summarize some of the key provisions for businesses.
NHTSA Proposes Easing Certain Regulatory Requirements for Self-Driving Vehicles
On March 17, 2020, the National Highway Traffic Safety Administration (NHTSA) published a notice of proposed rulemaking (NPRM) aiming to remove unnecessary barriers to vehicles equipped with automated driving systems (ADS) and proposing to amend and modernize 11 of its Federal Motor Vehicle Safety Standards (FMVSS) related to crashworthiness and occupant protection.
FMCSA Expands Emergency Regulatory Relief for Truck Transportation of Essential Supplies, Equipment, and Persons
Key Notes:
- Certain regulations pertaining to commercial motor vehicle drivers and motor carriers are suspended to facilitate COVID-19 emergency relief efforts.
- The suspension applies to transportation of essential supplies, equipment, and persons, including shipments of medical supplies, community safety and sanitation supplies, food, and the raw materials used to produce these items.
- The suspension ends upon termination of the COVID-19 emergency or April 12, 2020, whichever is earlier.
On March 18, 2020, the Federal Motor Carrier Safety Administration (FMCSA) issued an Expanded Emergency Declaration suspending enforcement of its hours-of-service, driver qualification, and other regulations set forth at 49 C.F.R. Parts 390-399 for commercial motor carriers and drivers providing direct assistance to emergency relief efforts.
COVID-19: DOL Reminds Employers of Their Duties Under the FMLA, ADA and FLSA
In addition to guidance given to employers by the Occupational Health and Safety Administration, the U.S. Department of Labor’s Wage & Hour Division issued several reminders to employers regarding their duties under the Family and Medical Leave Act, Americans with Disabilities Act, and Fair Labor Standards Act.
In This Section
Our Legal Advisor
Karyn A. BoothKaryn is a partner and leader of the firm’s Transportation practice group. Based upon interviews with clients and peers, Chambers USA has recognized Karyn as one of the leading lawyers nationwide who represent shippers in rail transportation matters, and she was also identified for her work in road transportation matters. Karyn represents multinational corporations, trade associations, and transportation intermediaries, such as 3PLs, NVOCCs, freight forwarders and brokers, in domestic and international matters involving multimodal transportation and logistics services. Her practice covers the carriage of goods by rail, motor, vessel and air carriers. Karyn serves as the general counsel to The National Industrial Transportation League, the nation’s oldest and largest shipper organization. Karyn’s practice includes a full range of services with a focus on regulatory compliance and counseling; proceedings before the Surface Transportation Board (STB), Department of Transportation (DOT), Federal Motor Carrier Safety Administration (FMCSA), Federal Railroad Administration (FRA), Federal Maritime Commission (FMC), Pipeline and Hazardous Materials Safety Administration (PHMSA), Department of Homeland Security (DHS), Transportation Security Administration (TSA), Customs and Border Protection (CBP), and Federal Aviation Administration (FAA); transportation contracting; transportation security; legislation; and litigation/arbitration of transportation-related disputes.
February 2020
Publications SHARE 0 Add to eBriefcase The Coronavirus Threatens U.S. Company Supply Chains - Practical Tips and Contract Considerations to Manage Risk
Key Notes:
- The risk of coronavirus transmission from imported goods is currently considered highly unlikely
- U.S. businesses should engage in contingency planning, including evaluating alternative sourcing and the potential future need for increased transportation capacity
- Disruptions may affect contracts between U.S. businesses and Chinese suppliers, and U.S. businesses and buyers of finished goods
- Transportation contracts should be reviewed for minimum volume commitments and capacity protections
There are growing concerns that the Coronavirus (“COVID-19”) outbreak will have a serious negative impact on the global economy. U.S. businesses that depend on Chinese suppliers may soon be facing product shortages and supply chain disruptions. This bulletin (i) addresses whether U.S. businesses need to be concerned about the possible exposure to the virus from goods imported from China, and (ii) identifies contractual considerations to help companies manage potential supply chain risks based on their relationships with Chinese suppliers, customers and transportation providers.
July 2019
FMCSA Seeks Information on CMV Driver Detention Times During Loading and Unloading
Last month, the Federal Motor Carrier Safety Administration (FMCSA) issued a request for information concerning commercial motor vehicle (CMV) driver detention times during loading and unloading. The request for information follows several studies examining the issue of CMV driver delays during loading and unloading and their impact on roadway safety and the economy.
February 2019
FAA to Seek Comments on Measures to Address Drone Risks
Key Notes:
- The FAA will be seeking input on measures to mitigate public safety and national security risks presented by drones.
- Recent events demonstrate that drones pose safety and security risks to airports, sporting events, and other sensitive infrastructure and public events.
- Entities concerned about the risks of drone operations or about how safety and security limitations would impact their drone operations should submit comments.
The Federal Aviation Administration (FAA) recently announced that it would solicit comments on regulatory measures to address public safety and national security risks posed by drones, which could result in the FAA limiting drone use or imposing onerous drone design standards. Thus, drone users and manufacturers, as well as businesses and other entities for which drones pose a safety risk, such as airports and power plants, should consider providing feedback to the FAA on appropriate measures to mitigate drone safety and security risks.
January 2019
FMC Issues Final Report on Demurrage and Detention Practices
In December 2018, the Federal Maritime Commission (FMC) issued its Final Report in Fact Finding Investigation No. 28: “Conditions and Practices Relating to Detention, Demurrage, and Free Time in International Oceanborne Commerce” (Final Report). In it, the Fact Finding Officer, Commissioner Rebecca F. Dye, sets forth her findings throughout the investigation’s phases and makes recommendations related to demurrage and detention practices that would improve the performance of the U.S. freight delivery system.
Past Items
FMCSA NPRM on Motor Carrier Safety Fitness Determinations
The Federal Motor Carrier Safety Administration (“FMCSA”) published a Notice of Proposed Rulemaking (“NPRM”) in the Federal Register on January 21, 2016 setting forth proposed rules for determining the safety fitness rating for interstate motor carriers that is based on on-road safety data and the results of safety investigations. Comments from interested parties are due on March 21, 2016 and reply comments are due on April 20, 2016.
FMCSA Prohibits Coercion of CMV Drivers, Extends Jurisdiction
On November 27, 2015, the Federal Motor Carrier Safety Administration (“FMCSA”) issued a final rule prohibiting motor carriers, shippers, receivers, and transportation intermediaries from coercing drivers to operate commercial motor vehicles (“CMV”) in violation of the FMCSA safety regulations and the federal Hazardous Materials Regulations (“HMR”), and prohibiting operators of CMVs from coercing drivers to violate certain FMCSA commercial regulations. The rule includes procedures for a driver to report an incident of coercion and for investigation and action by FMCSA in response to such reports.
FMC Actions on Port Congestion
In the fall of 2014, the US Federal Maritime Commission (“FMC”) conducted four forums “to hear firsthand the problems that stakeholders in the U.S. intermodal system were facing as a result of problems brought on by contemporary developments in liner shipping.” The forums were located at the Ports of Los Angeles, Baltimore, Charleston, and New Orleans, respectively, and resulted in the publication of two FMC reports. On April 3, 2015, the FMC released a Commission’s staff report specifically addressing detention, demurrage and free-time. Subsequently, in July, the FMC issued a more thorough report discussing the major themes arising from the fall 2014 port congestion forums. These reports are summarized briefly below. | https://www.hpclcnet.org/legal-updates/ |
Through the years, human culture has experienced many evolutions – reacting and adapting to new situations, new ideas, or new innovations. Human’s artistic outlets too have evolved, changing with man’s expression – a mirror with which we reflect ourselves. We look to the past to learn, but we look to the future to grow.
In “[music]x(evolution)” we explore the changes over time that man and music have experienced from Ancient History (rhythm) to The Classical Era (melody) to Modern Time (electronics). Musical themes evolve and expand as additional layers of responsibility are incorporated into the ensemble’s efforts. The diverse music can be enhanced via visual evolution of posture, technique and form from the rudimentary basics to advanced modern performance. Written into “Evolution” are opportunities to diversify the instrumentation or voices available within the ensemble’s sound. Potential props include standing drums, backdrops, and lights.
This production was originally written for that odd year where you have a really strong battery coupled with a more A class level front ensemble. Additional difficulty versions available on request.
“[music]x(evolution)” was originally commissioned for Mechanicsburg HS (Mechanicsburg, PA) for the winter of 2018 and was an WGI A Class Finalist. | https://front2backmusic.com/show/evolution/ |
It’s a great question! Your subscribers have busy lives and the last thing you want is to send them your newsletter while they are preoccupied with work or dreaming about their weekend.
Ideally, you want your readers to have enough time and mental space to read your newsletter and click through to more content if that’s part of your strategy.
Does such a time exist when your readers are perfectly primed to receive and read your email?
And, if “the perfect time” does exist, wouldn’t everyone send their newsletter at that exact time (which would completely defeat the purpose)?
Ok, before we get too philosophical, let’s take a look at the current approaches and dive into our data to help you figure out this million dollar question.
Most people put themselves in the shoes of their subscribers to help them figure out the best day of the week to send. When you lack data, this is a perfectly logical approach. Close your eyes and think about the daily schedules and people’s mindsets throughout the week.
The week usually plays out like this:
People are super busy adjusting back to the work week and they prioritize work emails. Because of this emphasis on work, people think Monday is not a good day to send your newsletter.
Most people believe Tuesday, Wednesday and Thursday are good days as people are settled in their work routine and constantly checking their inboxes.
On Friday, everyone is dreaming about the weekend and rushing to finish their work so they can go offline. From my experience, Friday afternoon might be great timing, especially right after lunch. People usually don’t start new tasks and have time to read the inbox.
The weekend is tricky. Some cultures, like the French and Italians, rarely check their inbox when they don’t work. On the other hand, Sunday evenings might be an ideal time in America, as they tend to get ready for the work week that night. Also, people get fewer emails on weekends, so it could be easier for you to get their attention.
By logically going through the days of the week, you can make some guesses, but it’s still not completely clear when to send your emails.
We decided to go a step further and dive into our data compiled from millions of campaigns.
We love crunching the numbers, and we’re big believers in analyzing data from a human perspective. We analyzed millions of MailerLite campaigns to find some insights that we could pair with our real-world experience to see if there is, in fact, an ideal sending window.
Here are the results. We'll include our analysis below the charts.
Email marketers tend to send more newsletters on workdays. The interesting insight here is that open rates don't differ much throughout the week, especially in the US and Canada.
While Saturday has a lower send volume, it also looks like it's the worst day for opens. Friday may be the best option, especially if you have a global audience.
These graphs show the time of the day when people open their emails and click during each hour of the day. For Tuesdays and Fridays, the highest percentage of people open their emails at 11 AM, yet the highest amount of clicks occurs at 9 AM.
The high volume of clicks at 9 AM suggests that people select the emails they find most relevant or interesting first thing in the morning. They engage with certain emails and save the rest for later.
Surprisingly, Sunday at 9 AM is when people both open their emails and click through.
Finding the best day and time from the data might not be the only variable when thinking about sending your emails. Look at your campaign holistically to see if things like your marketing goals, messaging and target audience influence the best time to send your email.
Think about what you want your reader to do when they get the email. Read it? Click through and make a purchase? Or maybe reply?
Here are 3 types of goals that inform your sending times.
Do you want your readers' full attention where they click through for a longer experience? Consider sending your newsletters either on Friday afternoon, in the evenings or even on the weekend. It’s easier to capture attention when fewer or no emails are sent.
If you are happy with readers quickly scanning your newsletter just to keep your brand top of mind, send it anytime during the weekday so your subscribers can view and move on quickly. This is the time when people read emails methodically throughout the day but only react to the important ones.
Finally, if your goal is to remind people about an event or a webinar, send a follow-up email a day before the event and then resend it an hour before the event to make sure people remember it.
Different types of content are better sent at certain times of the day. The most obvious example is a news and media email. If there’s something newsworthy, you send it as soon as possible.
Newsletters promoting B2B services or products perform better during working hours when people are in the work mindset. Whereas a travel agency or an author can capture more attention if they send emails after work hours or on the weekend.
Think about the type of message you are sending and time it appropriately.
Did you know that the ‘best time to send an email’ data is based mostly on desktop users? Check which type of device your subscribers use to open your newsletters. Mobile users tend to be more active late in the evening and on the weekends.
Or, maybe your primary audience is not a 9-5 business person, but rather a remote worker, a stay-at-home mom or dad, or a retired worker. Use this to your advantage by understanding their email habits and sending them newsletters when they’re available to engage.
Once you’ve analyzed your data and determined a couple of key times to send your newsletters, take advantage of some of email marketing’s best-sending features. They’ll make your life much easier!
Here are 3 that you can start using today:
Most email opens happen within an hour of arrival. If your message is very important, use auto-resend to reengage your readers without lifting a finger. An email will resend automatically to everyone that didn’t open it the first time.
You can add a personal message to the second email to let people know it’s important. The goal is to make everyone feel like the second email was designed especially for them.
With MailerLite, you can check your Opens by Location to help you understand where your subscribers are located.
If your subscribers are located in different time zones, use Deliver by time zones to set your desired delivery time for every time zone. This way, some of your subscribers won’t receive an email at 3 AM. Each subscriber will see the email at the same time in their time zone.
In certain circumstances, you can use link triggers to send your subscribers emails at the perfect time. Instead of trying to figure out the best time to send an email to your entire email list, link triggers let your subscribers dictate when to send the email based on their actions.
You can place links in your content, emails or on your website that trigger an email to send automatically. When a subscriber clicks on the link, they activate an email that matches the topic your reader is into at that very moment.
While link triggers aren’t meant for every scenario like a weekly newsletter, they do work great when you want to deliver a specific message at the exact time your subscriber is thinking about a topic.
No matter what time you send your newsletter, a best practice is to keep it consistent.
We are all creatures of habit and subconsciously crave consistency. Send your emails on the same day every month, week or day. By keeping it consistent, you build trust and anticipation as your audience expects your content at a designated time.
For example, I look forward to Seth Godin’s email every day. I know that on Sunday, I’ll get Paul Jarvis’ thoughts and on Friday, I’ll get an email from Tim Ferriss. Not only do I love these newsletters, but there is a Pavlovian response to receiving these emails that brings me joy.
To tell you the truth, the sending time is a minor detail in the broader email marketing strategy. People open and read emails because of the content and the value you provide them.
If your deadline is coming up and you are not happy with your newsletter, wait until it's ready! It's way more important to deliver a quality experience than to make a specific sending date and time.
You want your readers to look out for your newsletters with excitement. When you can pull that off, the timing becomes less relevant. | https://www.mailerlite.com/blog/best-time-to-send-email |
This study examined the time course of recovery following resistance exercise sessions in the back squat, bench press, and deadlift. Twelve well-trained males (age: 24.5±3.8yrs, body mass: 84.01±15.44kg, training age: 7.1±4.2yrs) performed 4 sets to failure at 80% of a one-repetition maximum (1RM) on the squat, bench press, and deadlift in successive weeks. The bench press was always performed in week-2 with the squat and deadlift order counterbalanced between weeks 1 and 3. Indirect muscle damage and performance fatigue was assessed immediately prior to, post-exercise, and 24, 48, 72, and 96hrs post-exercise...
Objectives: The aim of this study was to explore the effects of laser acupuncture on improvement of recovery and muscle performance in delayed muscle soreness (DOMS) when applied before exercise. Methods: This randomized, blinded, and controlled study included healthy participants (n = 40) who were randomized into laser acupuncture and placebo groups. Laser acupuncture was applied to the Tianquan (PC2) and Chihtseh acupoints (LU5) at a dose of 36 J and energy density of 9...
Underfat individuals have been neglected as a malnourished population in terms of redox homeostasis. The aim of the present study was to evaluate the effect of body composition on redox homeostasis at rest and in response to exercise. Underfat, lean and overfat women, classified according to their BMI and body fat percentage, participated in the study and were subjected to an acute session of eccentric exercise. With regard to muscle function and damage, a significant group × time interaction was found for range of motion (P < ...
Background/Objective: In soccer the recovery time between matches is often not long enough for complete restoration. Insufficient recovery can result in reduced performance and a higher risk of injuries. The purpose of this study was to evaluate the potential of Deep Oscillation (DO) as a recovery method. Methods: In a randomized crossover study including 8 male soccer players (22 ± 3.3 years) the following parameters were evaluated directly before and 48 h after a fatiguing soccer-specific exercise: Maximum isokinetic strength of the leg and hip extensors and flexors (Con-Trex® Leg Press, Physiomed, Germany), rating of perceived exertion (RPE) during isokinetic testing (Borg scale 6-20), creatine kinase (CK) serum levels and Delayed Onset Muscle Soreness (DOMS; visual analogue scale 1-10)...
PURPOSE: Sudden and exhaustive exercise causes muscle damage accompanied by oxidative stress and inflammation, leading to muscle fatigue and soreness. AmLexin contains a standardized blend of extracts from the heartwood of Acacia catechu and the root bark of Morus alba, and is known to possess antioxidant and anti-inflammatory properties. The aim of this study was to investigate the effects of this proprietary blend supplementation on muscular pain and redox balance in healthy runners, in comparison to a placebo...
During the preseason, futsal players deal with large internal load, which may result in a reduction in physical performance. The aims of this study were to compare the session rating of perceived exertion training load (s-RPE TL) during the preseason between two teams; and to analyze the changes on the delayed-onset muscle soreness (DOMS), aerobic- and speed-power characteristics in players accumulating different s-RPE TL (Low (LTL) vs. High (HTL)). Twenty-eight players (Team A, n = 15; Team B, n = 13) were recruited...
Delayed-onset muscle soreness (DOMS) is a type of ultrastructural muscle injury. The manifestation of DOMS is caused by eccentric or unfamiliar forms of exercise. Clinical signs include reduced force capacities, increased painful restriction of movement, stiffness, swelling, and dysfunction of adjacent joints. Although DOMS is considered a mild type of injury, it is one of the most common reasons for compromised sportive performance. In the past few decades, many hypotheses have been developed to explain the aetiology of DOMS...
OBJECTIVE: Delayed-onset muscle soreness (DOMS) is a symptom of exercise-induced muscle injury that is commonly encountered in athletes and fitness enthusiasts. Vibration is being increasingly used to prevent or treat DOMS. We therefore carried out a meta-analysis to evaluate the effectiveness of vibration in patients with DOMS. METHOD: We searched nine databases for randomized controlled trials of vibration in DOMS, from the earliest date available to 30 May 2018...
STUDY DESIGN: Systematic review with meta-analysis. BACKGROUND: Needling has been shown to decrease pain in the short-term; however, its effects on muscle force production are unclear. OBJECTIVE: Evaluate the evidence regarding the comparative effects of needling on muscle force production. METHODS: An electronic search was performed using keywords related to needling. Methodological quality of articles was appraised and effect sizes calculated...
BACKGROUND: Sensing tissue acidosis is an important function of the somatosensory nervous system to response to noxious stimuli. MAIN BODY: In the pain clinic, acid or soreness sensation is a characteristic sensory phenotype of various acute and chronic pain syndromes, such as delayed onset muscle soreness, fibromyalgia, and radicular pain. However, soreness sensation is a sign of successful analgesia for acupuncture and noxipoint therapy. Thus, the nature of acid or soreness sensation is not always nociceptive (or painful) and could be anti-nociceptive...
Objective: The aim of the present study was to examine the effect of Cold Water Immersion (CWI) on the recovery of physical performance, hematological stress markers and perceived wellness (i.e., Hooper scores) following a simulated Mixed Martial Arts (MMA) competition. Methods: Participants completed two experimental sessions in a counter-balanced order (CWI or passive recovery for control condition: CON), after a simulated MMAs competition (3 × 5-min MMA rounds separated by 1-min of passive rest). During CWI, athletes were required to submerge their bodies, except the trunk, neck and head, in the seated position in a temperature-controlled bath (∼10°C) for 15-min...
OBJECTIVES: The symptomatic reduction of exercise-induced delayed onset muscle soreness (DOMS) is of great interest in the fields of Sports Medicine and Physical Therapy. At this time, few therapeutic interventions have proven their effectiveness. One of the most promising interventions is Compression Therapy. The object of this study is Tissue Flossing, a therapeutic short-term compression method and whether its post-exercise application can reduce the extent of perceived DOMS. DESIGN: randomized, controlled, and double-blind pilot cross-over trial...
OBJECTIVE: Measurement of skin temperature using infrared thermography has become popular in sports, and has been proposed as an indicator of exercise-induced muscle damage after exercise. However, the relationship between skin temperature and exercise-induced muscle damage is still unclear. Here we set out to investigate the relationship between skin temperature and exercise-induced muscle damage. APPROACH: Twenty untrained participants completed a protocol of exercise for calf muscles...
This paper aims to critically evaluate and thoroughly discuss the evidence on the topic of caffeine supplementation when performing resistance exercise, as well as provide practical guidelines for the ingestion of caffeine prior to resistance exercise. Based on the current evidence, it seems that caffeine increases both maximal strength and muscular endurance. Furthermore, power appears to be enhanced with caffeine supplementation, although this effect might, to a certain extent, be caffeine dose- and external load-dependent...
The purpose of this study was to investigate the involvement of neutrophil dynamics and function in exercise-induced muscle damage (EIMD) and delayed-onset muscle soreness (DOMS), and the effect of molecular hydrogen (H₂) intake on these parameters. Nine healthy and active young men performed H₂ and placebo bath trial in a crossover design. They carried out downhill running (-8% slope) for 30 min at a speed corresponding to 75~85% of peak oxygen uptake (VO₂peak). Subsequently, they repeated bathing for 20 min per day for one week...
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Below is a Definition of Queueing Theory, a glossary, and vocabulary. Knowing the concepts below will help you better understand these articles on Queues.
- Queue: A line (or buffer or inventory) feeding a number of servers
- Server: An operation fed by a queue.
- Arrival rate (λ): Mean number of arrivals per unit time (usually per hour or day).
- Service rate (μ): Mean number of customers that can be served at 100% utilization by each individual server per unit time (usually per hour or day). At the individual workstation level, the service rate will equal capacity.
- Channels (M): The number of parallel operations connected to an individual queue. For example, if each queue has 2 operations then it will have two channels.
- Utilization (u): A measure of how “busy” the system is. It is generally defined as the ratio of throughput to capacity. Note that u = λ/(Μμ) if λ < Μμ, i.e. the utilization is less than 100%. (Also, note that while the Greek letter μ— or mu— looks a bit like u, they are in fact two different variables.)
- Phase: A queue and its connected servers, or routes to a server.
- Balking: When a person, who would otherwise have entered a line, decides not to enter it.
- Reneging: When a person, who has entered a line, later decides to leave it without being served.
- Interarrival Time: The time between when one customer arrives at a queue and when the next customer arrives.
- Service Time: The time it takes for one particular server to complete a customer’s service. The average service time will be the same as the cycle time.
And now for some equations you’ll need to know.
- CV: The coefficient of variation. This is a measure of a random variable’s variability. For a random variable x, CVx is defined as Standard Deviation (x) x mean (x) CV = .
- CVIAT: The coefficient of variation of the interarrival time. The greater the CVIAT, the “lumpier” the arrival rate.
- CVST: The coefficient of variation of the service time. The smaller the CVST, the more “consistent” a server is.
- Lq: The average number of people in a line awaiting service.
- Wq: The average length of time a customer waits before being served.
After you have become familiar with the terms above, go ahead and read the articles on Queueing Theory. | https://opexlearning.com/resources/queueing-theory-definition/8033/ |
Since 2004, concerns and calls for greater quality assurance in experiential education have been published. The Accreditation Council for Pharmacy Education (ACPE) "Standards 2016" provide limited differentiation across the four required practice experiences, and, as such, schools interpret them differently. Both schools and accreditation site visit teams would benefit from a common set of guidance for the required Advanced Pharmacy Practice Experiences (APPEs), so that they can ensure consistency and quality in student experiences across practice sites. To address this need for greater standardization, a taskforce of the American Association of Colleges of Pharmacy (AACP) Experiential Education (EE) Section conducted a peer-reviewed, consensus-building process, including experiential faculty and staff across multiple colleges and schools of pharmacy, to determine a common set of elements that could be used to bring consistency to the experiences and expectations for student learning in practice. Over a two year period, the taskforce reviewed the relevant literature and then drafted and revised the elements through an iterative process which allowed for established EE consortia and members of the EE section to review the draft and provide input for revision. The resulting essential elements presented here can be used to guide faculty and staff within experiential education programs in their quality assurance processes in ensuring students receive consistent experience as part of their education prior to graduation.
RESUMEN
Introduction: The science of patient safety demonstrates that good communication is essential for effective interprofessional collaboration. Methods: We created a low-stakes, formative assessment with which medical students, pharmacy students, and nursing students could practice several of the Interprofessional Education Collaborative competencies. We aimed to enable students to practice collaborative care, respect for other disciplines, and shared accountability. Senior students from medicine, nursing, and pharmacy worked in teams to disclose a medical error to a standardized patient. The activity began with an icebreaker exercise wherein students learned about each other. Next, each team planned a strategy for error disclosure and collaboratively disclosed the error. Standardized patients evaluated the team's performance. Subsequently, students regrouped for a debriefing. The participating institutions administered a survey to their students. Results: In total, 1,151 students participated: 464 fourth-year students from the University of Houston College of Pharmacy, 450 third- and fourth-year students from Baylor College of Medicine, and 237 fourth-year students from Texas Woman's University Nelda C. Stark College of Nursing, all in Houston, Texas. Postsession survey data showed that students thought they achieved the relevant competencies. Students' understanding of the perspectives of the other two disciplines improved. Students found the simulation encounter and debriefing effective in helping them consider the contributions of other disciplines to patient care. Discussion: This interprofessional standardized patient activity enabled collaborative problem solving. The debriefing discussion broadened students' understanding of the expertise of the other disciplines and promoted shared accountability. Students found this activity engaging and effective.
RESUMEN
OBJECTIVE: The current literature has identified many motivating factors and barriers influencing pharmacy students' decision to apply for residency training. Despite a growing need for residency trained pharmacists to advance the profession, it is not clear why only about one in four pharmacy students decide to pursue a residency, and which of these factors have the most influence on student decision-making. The study examines the factors associated with pharmacy students' intention to apply for a postgraduate residency using the theory of planned behavior (TPB) framework. METHODS: Second and third-year students from four Texas pharmacy schools were surveyed using an online questionnaire based on the TPB. Descriptive statistics and multiple linear regression analyses were utilized to assess the study objectives. RESULTS: A total of 251 completed responses were received. Attitude, subjective norms (SN), and perceived behavioral control (PBC) were significant predictors of intention to apply for a pharmacy residency (ß = 0.32, 0.58, and 0.36, respectively, p < 0.001). Attending ASHP's midyear meeting or other residency showcase was a significant predictor of intention (ß = 0.71, p = 0.006). Additional significant predictors of intention include believing a residency would increase confidence in practicing pharmacy (ß = 0.36, p < 0.001) and help achieve career goals (ß = 0.16, p < 0.02); the social influence of faculty members (ß = 0.10, p = 0.003) and family (ß = 0.08, p = 0.02); believing financial obligations (ß = 0.20, p = 0.006), feeling afraid of the competition and/or not matching (ß = 0.24, p < 0.001), needing to relocate (ß = 0.09, p = 0.04), and the lengthy application and/or interview process (ß = 0.12, p = 0.04) would make it more difficult to apply for a residency. CONCLUSIONS: The TPB model was useful in predicting pharmacy students' intention to apply for a residency, and all TPB constructs were significant predictors. Therefore, interventions that target students' attitude, SN, and PBC may be valuable to increase their intention, especially the specific beliefs identified to significantly predict intention. Future research into methods in which these motivating factors can be encouraged and perceived barriers can be addressed by pharmacy stakeholders will increase interest and participation in residency training. | https://pesquisa.bvsalud.org/portal/?lang=es&q=au:%22Ordonez,%20Nancy%22 |
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An influential theory of long-term memory, in which new episodic learning is dependent upon the integrity of semantic memory, predicts that a double dissociation between episodic and semantic memory is not possible in new learning. Contrary to this view, we found, in two separate experiments, that patients with impaired semantic memory showed relatively preserved performance on tests of recognition memory if the stimuli were perceptually identical between learning and test. A significant effect of semantic memory was only seen when a perceptual manipulation was introduced in the episodic task. To account for these findings, we propose a revision to current models of long-term memory, in which sensory/perceptual information and semantic memory work in concert to support new learning.
National Center for
Biotechnology Information, | https://www.ncbi.nlm.nih.gov/pubmed/10678697?dopt=Abstract |
The MistraPharma research program proposes 10 recommendations for improving environmental risk assessment of human pharmaceuticals. The recommendations are based on up-to-date scientific knowledge concerning pharmaceuticals’ effects and presence in the environment, as well as experiences from the REACH-framework that regulates industrial chemicals.
Download the policy brief here or read abour our recommendations below.
Require environmental risk assessment also for products
put on the market before 2006
put on the market before 2006
We recommend that environmental risk assessments are performed also on pharmaceutical products approved before the European Medicines Agency’s guideline came into force. There is simply no scientific evidence that products put on the market before 2006 are of less environmental concern than new products.
Perform only one environmental risk assessment per pharmaceutical substance
We recommend that pharmaceutical companies that produce or import the same pharmaceutical substances submit a joint environmental risk assessment instead of submitting one per company. This would provide decision makers with coherent information, avoid duplication of work and reduce animal testing.
Perform mixture toxicity assessments on pharmaceutical substance with similar modes of action
We recommend that environmental risk assessments also consider the total exposure for groups of pharmaceutical substances with similar modes of action. This would enable a more accurate assessment of the environmental risks.
Include environmental risks in the risk-benefit analysis
We recommend that environmental risks are included in the risk-benefit analysis when a product is considered for market authorization. This would make the assessment of risks associated with the use of pharmaceuticals more complete and hence more accurate.
Include data on emissions from production of pharmaceutical substances
We recommend that the risk associated with discharges from manufacturing sites is included in environmental risk assessments. This would enhance the relevance of the assessments by including the part of the product lifecycle responsible for the highest environmental concentrations detected.
Add requirements to assess the risk for development of antibiotic resistance
We recommend that information that enables assessment of the risk for increased antibiotic resistance development is included in the environmental risk assessment for antibiotic substances. This would provide a more accurate picture of the risks associated with environmental occurrence of antibiotics.
Refine the tiered approach
We recommend that the test approach is refined to include pharmacological and toxicological data from the drug discovery process, as well as bioconcentration data. This would improve the prioritization process for further testing so that ecotoxicity testing is focused on the most problematic substances and the most relevant test organisms.
Mandate use of all available ecotoxicity studies
Require review of the environmental risk assessments at regular intervals
Increase transparency
We recommend that environmental risk assessments and information about manufacturing sites are made publicly available. This would enable use of that information for other purposes such as research and external evaluation, and it would encourage companies to take more environmental responsibility throughout their supply chains. | https://www.mistrapharma.se/outcomes/policy-brief-27166372 |
A proposition is any statement with a truth value. A premise is a proposition that is being used in a particular way.
How is a premise different from a proposition?
As verbs the difference between premise and proposition
is that premise is to state or assume something as a proposition to an argument while proposition is to propose a plan to (someone).
How do you identify a premise?
The foolproof way to do this is to ask yourself what the author of the argument is trying to get you to believe. The answer to this question is the conclusion. There must also be at least one reason and possibly many. These are your premises.
How will you identify if the proposition is a conclusion or premise?
If it’s being offered as a reason to believe another claim, then it’s functioning as a premise. If it’s expressing the main point of the argument, what the argument is trying to persuade you to accept, then it’s the conclusion.
What is an example of a premise?
The definition of a premise is a previous statement that an argument is based or how an outcome was decided. An example of premise is a couple seeing a movie chosen by one, because they saw a movie chosen by the other last week.
What is a proposition example?
A proposition is a declarative sentence that is either true or false (but not both). For instance, the following are propositions: “Paris is in France” (true), “London is in Denmark” (false), “2 < 4” (true), “4 = 7 (false)”.
Are premises and conclusions both propositions?
The premises of an argument are those statements or propositions in it that are intended to provide the support or evidence. The conclusion of an argument is that statement or proposition for which the premises are intended to provide support. (In short, it is the point the argument is trying to make.)
What is the difference between premises and conclusion?
A premise is a statement in an argument that provides evidence or reasons to form a conclusion. It contains the information that leads your audience to believe that your argument is true. An argument can have one or more premises.
What is a premise in logic?
Premise: Proposition used as evidence in an argument. Conclusion: Logical result of the relationship between the premises. Conclusions serve as the thesis of the argument. Argument: The assertion of a conclusion based on logical premises.
What are some examples of premise indicator words?
Conclusion and premise indicators are words that are used to make clear which statements are premises and which statements are conclusions in arguments. Here’s a list of the most common ones.
What is an argument?
|Conclusion indicators||Premise indicators|
|Hence||Supposing that|
|Consequently||Assuming that|
|Ergo||Given that|
What is proposition explain?
noun. the act of offering or suggesting something to be considered, accepted, adopted, or done. a plan or scheme proposed. an offer of terms for a transaction, as in business. a thing, matter, or person considered as something to be dealt with or encountered: Keeping diplomatic channels open is a serious proposition.
What is simple proposition?
Simple propositions are declarative sentences which do not contain a connective. The restriction to declarative sentences is important. In propositional logic each proposition, simple or complex, must be capable of being either true or false. So we won’t count questions or commands, for example, as simple propositions.
How do you use proposition in a sentence?
Examples of proposition in a Sentence
Noun He made an attractive business proposition. The other company rejected their proposition. Her theory rejects the basic proposition that humans evolved from apes. If we accept proposition “A” as true, then we must accept proposition “B” as false.
What are 5 examples of prepositions?
Examples of common prepositions used in sentences include:
- He sat on the chair.
- There is some milk in the fridge.
- She was hiding under the table.
- The cat jumped off the counter.
- He drove over the bridge.
- She lost her ring at the beach.
- The book belongs to Anthony.
- They were sitting by the tree.
What are the 4 types of preposition?
Types of Prepositions
- Simple Preposition. When a preposition consists of one word is called single or simple preposition. …
- Double Preposition. When a preposition consists of more than one word, it is called double preposition. …
- Compound Preposition. …
- Participle Preposition. …
- Disguised Prepositions. …
- Phrase Prepositions.
What are the rules of prepositions?
Six Preposition Rules
- A preposition must have an object. All prepositions have objects. …
- pre-position means place before. …
- A pronoun following a preposition should be in object form. …
- Preposition forms. …
- 5. to preposition and to infinitive are not the same. …
- The golden preposition rule.
What is the easiest way to learn prepositions?
Sort by preposition and write complete sentences from the text. Once you’ve highlighted the prepositions in your reading passage, grab a notebook and write down each separate preposition—”in,” “under,” “at,” and so on—at the top of its own page. Then, write down each sentence in the text that uses that preposition.
How do you memorize prepositions?
Around at before beside between against within without beneath through during under in into over of off – toward upon near for from except.
What are the 8 types of preposition?
The 8 types of prepositions in English grammar with examples include prepositions of time, place, movement, manner, agent, measure, source and possession.
What are the 3 types of prepositions?
Page 1
- Place of prepositions – show WHERE something HAPPENS • Sanny was sitting under a tree. • …
- Time of prepositions – show WHEN something happens • School starts at nine o’clock. • …
- Direction of prepositions – show WHERE something is GOING • The boys chased after each other. • …
- COMMON PREPOSITIONS. across.
How many prepositions are in English grammar?
150
Prepositions are common in the English language. There are about 150 used with the most common being: above, across, against, along, among, around, at, before, behind, below, beneath, beside, between, by, down, from, in, into, near, of, off, on, to, toward, under, upon, with and within.
What are the 6 types of preposition?
There are six types of prepositions.
- Prepositions for Time.
- Prepositions for Place.
- Prepositions for Direction.
- Prepositions for Agency.
- Prepositions for Instruments.
- Prepositional Verb.
What are the 10 types of preposition?
There are some prepositions which are common in every type of preposition as they function in a versatile way.
- Simple Preposition.
- Double Preposition.
- Compound Preposition.
- Participle Preposition.
- Disguised Preposition.
- Detached Preposition.
- Prepositions of Time.
- Prepositions of Place and Direction.
What is participle preposition?
Participle prepositions (also called participial prepositions) are participles that are used as prepositions. Remember participles are words that are used to make compound verbs, nouns, adjectives, and in some cases used as a preposition. They are formed by -ing, -ed, -en, etc. | https://goodmancoaching.nl/how-does-one-differentiate-between-premise-and-proposition/ |
Because of everyone’s extraordinary satisfaction with Mia (Liu Manqiong), the director of the Human Resources Department of Aurous Hill University was afraid that there would be any changes in this matter, so he wrote a line on a white paper during the interview, which read, “Too excellent!” , There must be a lot of schools staring at it, and I suggest that re-examination be waived and be confirmed as soon as possible!”
Although this lady is the person in charge of the human resources department, the decision to hire or not is in the hands of one of the vice principals.
The vice-principal saw the words on the paper, and wrote two words “Agree.”
Then, he pushed the paper in front of another interviewer, and the other interviewer quickly followed “Agree”. wrote a “+1”.
So, the three of them passed messages to each other by means of notes, and made an unprecedented decision.
At the end of the interview, the woman who was about forty years old stood up, smiled and said to Mia (Liu Manqiong), “Miss Liu, since your resume and qualifications are very suitable for our positioning of young talents, after the unanimous decision of the three of us, , now I can give you a clear answer, you are hired!”
Mia (Liu Manqiong) asked in surprise, “I don’t need to take the re-examination or written examination?”
The woman smiled and said, “No, although our normal recruitment process requires One-sided, two-sided and written test, but only one part is an essential link. Both the second-sided and written test can be adjusted according to the actual situation. The three of us feel that your situation in all aspects is very suitable for us. needs, and you live in Hong Kong Island now, we don’t need to make you go back and forth again and again. If you are sure that you are willing to stay in our school to teach, then please adjust your itinerary and stay in Aurous Hill for one more day. The three of us have completed the recruitment process today and prepared the signing materials, and we can formally sign the contract tomorrow morning.”
After that, she looked at Mia (Liu Manqiong) expectantly and asked, “What does Miss Liu think? Now we can also sign the contract.” Are you sure, or do you want to go back and think about it?”
Actually, these three interviewers were more nervous than Mia (Liu Manqiong).
Mainland almostAll the double-first-class universities are very interested in talents like Mia (Liu Manqiong), and almost all of them have similar young talent introduction plans. With Mia (Liu Manqiong)’s conditions, interviews for the position of ordinary lecturers, any university in the mainland, as long as it is fair and just There is absolutely no difficulty in competing.
They also don’t know why Mia (Liu Manqiong) chose to come to Aurous Hill to develop as a native of Hong Kong Island. They are worried that Mia (Liu Manqiong) has no special feelings for Aurous Hill. Tomorrow Mia (Liu Manqiong) will sign to another school.
Mia (Liu Manqiong) didn’t know what the three of them were thinking. She originally thought that the interview would be a one-stop process, but she didn’t expect to get the offer from the other party in one step. This was naturally the most ideal result for her, so she said without thinking “I’m fine!” The
other three were also relieved, and the woman said, “Since everyone is fine, let’s sign the contract here at nine o’clock tomorrow morning. I will prepare the contract in advance. Just bring the relevant documents, and you don’t need to prepare copies, we can make copies everywhere.”
“Okay!” Mia (Liu Manqiong) said with a smile, “Thank you for your approval, I will come here on time tomorrow morning.”
What did the woman think of and said to her Said, “By the way, Ms. Liu, in view of your special situation, we can provide you with a teacher’s apartment. I can show you around tomorrow, or you can choose a room first.”
Mia (Liu Manqiong) originally wanted to say that she planned to buy a suite in Aurous Hill, It will not take up school resources.
But after thinking about it, housing prices in Aurous Hill are not cheap. I have just graduated and bought a house before I have a formal job. In the eyes of others, it may be too ostentatious. Why not accept the apartment arranged by the school, and then quietly Buy a suite. | https://m.informativestore.com/the-amazing-son-in-law-chapter-5440/ |
Hello, dear readers. Thank you all for the consistent support. Today, I will like to conclude my series on NERVOUS SYSTEM by starting with the discussion of the Parkinson’s Disease in relationship to a special part of the human brain.
Parkinson’s disease
Parkinson’s disease is a disease in which the death of a small number of cells in the basal ganglia leads to an inability to select and initiate patterns of movement. The exact symptoms vary, but generally Parkinson’s patients have an increased rigidity in their muscles, usually accompanied by a tremor. This leads to slowness of movement, poor balance and speech problems. This shows the importance of the basal ganglia – without them, the body is incapable of normal movement even though most of the motor system is intact.
Quite what triggers the onset of Parkinson’s is still not clear. It appears to be due to a genetic predisposition and an as yet unidentified environmental trigger. What is known is that the underlying cause is an inability to produce dopamine, a neurotransmitter that has a number of functions, including enabling us to move smoothly and normally.
Some relief from Parkinson’s has been achieved by treatment with levadopa, a precursor that is transformed into active dopamine in the brain. Dopamine reduces the symptoms – often spectacularly so – but is not a cure. Trials involving transplants of dopamine-producing cells from fetal or animal sources also show promise, but it is still early days.
Electroencephalograms and brain stem death
In 1929, the German scientist Berger discovered that the brain produces electrical activity that can be measured by electrodes placed on the scalp. The machine that measures brain waves produces a trace encephalogram, or EEG. It was discovered that the patterns of brain waves recorded on the EEG change in different situations, particularly with levels of consciousness. The four basic types of brain waves are alpha, beta, theta and delta. The absence of any electrical activity from the brain of a patient indicates brain stem death. The absence of breathing movements and electrical brain activity is the clinical definition of death. EEG traces tell us little about how the brain works, but they are useful for locating and diagnosing the various types of epilepsy, sleep disorders and brain tumours.
Critical windows in brain development
We know that the way we develop is largely due to our genes, but it is becoming increasingly apparent that our nervous system does not develop properly without the right sensory stimulation at the right time.
There are critical windows or critical periods when a young animal must have sensory or motor input if ‘normal’ development is to take place. During a critical window, neural connections are made between sense organs and vital areas of the brain. These critical periods exist in virtually all species, from fruit flies to humans. If a skill is not acquired during a critical period, the acquisition of that skill in later life will be much harder, if not impossible.
For example, kittens are born with their eyes shut and if they do not open at the right time, permanent blindness results. There is nothing wrong with the eyes themselves, but the visual cortex where the incoming information is processed fails to develop properly.
Key work in this area came from the experiments of David Hubel and Torsten Nils Wiesel, who won the 1981 Nobel Prize for Physiology for their work. They found that if a kitten is deprived of light in one eye during the critical period, there is only partial development of the visual cortex. They can only see with one eye, and although they can perceive detail well, there is very poor judgement of distance. Electrical studies of the visual cortex showed that only about one-seventh of the visual cortex develops, compared with full development following input from two eyes.
To further refine their research, kittens were placed in striped tubes (where the stripes were horizontal for some kittens and vertical for others) or were fitted with goggles that presented vertical stripes to one eye and horizontal stripes to the other.
When tested a few months later, after removal of the striped tubes and goggles, the kittens seemed to be blind to stripes with the opposite orientation to those they saw during rearing. Most of the cortical cells of the cats reared with horizontal stimuli subsequently responded strongly to horizontal stimuli and hardly at all to vertical stimuli. The opposite was true of the vertical stimuli-reared cats.
This has implications for children who have injured an eye, and have it covered with a bandage or patch. If the injury occurs during the critical window, the visual cortex will not develop properly and vision will be impaired.
The ethics of animal research
Few subjects in science cause so much fierce debate as that of animal rights. Was it right for Hubel and Wiesel to experiment on kittens and make them partially sighted for life? Was it right for Banting and Best to experiment on dogs giving them diabetes and ultimately killing them?
Most people believe that humans have certain rights, but it is very difficult to apply these ideals to animals. We can’t do medical research on people without their consent, but it is obviously not possible for animals to give their consent. It makes more sense to consider animal welfare rather than animal rights. Most people think that animals should have clean water, food, exercise/stimulation and access to veterinary care.
Some people think that to use animals in experiments is being speciesist (in the same sense as racist or sexist). They argue that by experimenting we are automatically assuming that human life is more valuable than that of any other species. Others argue that it is very difficult to draw the line, and that if the argument is taken to ts logical conclusion, we shouldn’t ever kill pests like rats or even locusts.
How much do animals suffer?
There is a general consensus among scientists that humane treatment should be given to those organisms whose nervous system is advanced enough to appreciate it. The argument is that pulling the legs off a daddy-long-legs or boiling a lobster (which, for all its size, is no more sophisticated than an insect) is not inflicting pain because these animals do not have a nervous system advanced enough to perceive pain and suffering.
No country in the European Union is allowed to experiment on vertebrates in medical experiments if non-vertebrate alternatives exist. If there is no alternative – as would be the case with the kittens – animals can be used provided that strict guidelines are followed. Many drugs – sleeping pills, for example, can only be tested using an intact, conscious mammal.
Scientists commonly take a utilitarian, or ‘greater good’, approach, meaning that the right course of action is the one that leads to the greatest overall benefit, i.e. the least suffering and loss of life in the long-term. A utilitarian framework is in place today, allowing a certain amount of animal experimentation provided that the overall benefits are expected to be significantly greater than the expected suffering and or loss of life.
Epilepsy and the two sides of the brain
Epilepsy is a common disorder of the brain, Symptoms range in severity from a mild loss of concentration, known as an absence or petit mal, to full-blown convulsive fits (grand mal) in which the subject blacks out and falls to the floor. These can be dangerous if the sufferer lashes out – injuring himself and others – or bites his own tongue.
The underlying cause of epilepsy is random, uncontrolled activity of some cells in the brain. This chaotic activity In both sensory and motor nerves causes patients to see and hear a variety of strange things, such as flashing lights and bells, while muscles jerk uncontrollably.
A diagnosis of epilepsy can be confirmed and studied using an EEG machine.It can be used to show a trace from a person during an epileptic seizure: you can compare it with the normal readings.
Epilepsy can often be controlled successfully by drugs. However, in extreme cases, the condition is treated by brain surgery, and one such operation has given a fascinating insight into the workings of the brain. The cerebral hemispheres have been described as two separate brains, and in order to work effectively as a whole, the two halves must communicate.
The bridge between the two halves is known as the corpus callosum. Neuroscientists discovered that the corpus callosum was involved in the spread of epileptic seizures. In a seemingly drastic operation, surgeons sever most of the corpus callosum. This often causes the seizures to be less intense and dangerous. However, there are other amazing consequences. Initially, subjects appear to be perfectly normal: they can talk and read, and have no problems in recognizing the world around them. However, if they close their right eye, and was given a familiar object such as a comb, they cannot put a name to it. Open the other eye, however, and ‘Ah, it's a comb!’
The same happens with words. If a word such as ‘TIGER’ is looked at it with the left eye only, the patient can’t read it. If they open the right eye, they can read the word immediately. This is because the left eye supplies information to the right side of the brain, and that is not where the language centre is situated. The right eye supplies information to the left side of the brain, to the language-processing neurons.
From studies of split-brain patients, and other studies, it appears that different sides of the brain have different functions. The left hemisphere contains the language centre, and the three Rs – reading, writing and ‘rithmetic. The right side, in contrast, is responsible for our imagination and sense of humour. It can also appreciate form, geometry and music. It cannot, however, put words to things. If the right hemisphere needs a word, it has to put in a call to the left side, via the corpus callosum.
Split-brain patients do not experience the symptoms tor ever. Within a few months the right hemisphere develops more language skills and can function on its own. It has even been suggested that split-brain patients could read two books simultaneously, one with each eye!
THE PERIPHERAL NERVOUS SYSTEM
The nerves of the peripheral nervous system behave like major road systems, carrying traffic in and out of the central nervous system. Afferent nerves, also called sensory nerves, carry information from sensory receptors into the CNS. Efferent nerves, also called motor nerves, carry information from the CNS out to effector organs. The efferent system can be further subdivided into the somatic and autonomic systems. These differ in their function, rather than their structure or position in the body. Afferent means ‘incoming’ while efferent means ‘outgoing’. You can refer to ‘afferent nerves’ or ‘efferent blood vessels’, for example.
The somatic nervous system
The somatic nervous system contains both afferent and efferent nerves. It receives and processes information from receptors in the skin, voluntary muscles, tendons, joints, eyes, tongue, nose and ears, giving an organism the sensations of touch, pain, heat, cold, balance, sight, taste, smell and sound. It also controls voluntary actions such as the movement of arms and legs.
The autonomic nervous system
The autonomic nervous system (ANS) consists of two sets of involuntary nerves that generally act antagonistically – these are the sympathetic and parasympathetic systems and they have opposing effects. The system is entirely motor, made up of efferent nerves only. It does not carry sensory information: feedback from muscles and glands travels via the somatic system.
The ANS controls basic ‘housekeeping’ functions such as heart rate, breathing, digestion and blood flow. Heart rate, for example, can increase or decrease. So, while the sympathetic system increases heart rate, the parasympathetic system lowers it. Generally, the sympathetic system has a stimulatory effect, and prepares the body for action, while the parasympathetic system returns body functions to normal.
Normally, the activity of both systems is balanced. But if the body is stressed, then the ‘fight or flight’ reactions of the sympathetic nervous system take over, causing an increase in heart rate, faster breathing, an increase in blood pressure and an increase in blood sugar level. This makes the body ready for sudden strenuous activity. When the emergency is over, the parasympathetic system takes over. It decreases the heart and breathing rates and diverts blood supply back to housekeeping’ activities such as digestion and food absorption. The actions of the parasympathetic nervous system have been described as ‘feed or breed’ because parasympathetic stimulation leads to increased blood flow and peristalsis in the intestines, and sexual responses such as gaining an erection.
The autonomic system was originally thought to be independent of the rest of the nervous system, hence the term autonomic, meaning ‘on its own’ or ‘self governing’. Now we appreciate that it is not autonomous, but is regulated by areas within the central nervous system, including the hypothalamus, cerebral cortex and the medulla oblongata.
Do we have any conscious control over the autonomic nervous system?
The answer is yes, in some cases. As children we became potty trained when we learned conscious control over the muscular valves (sphincters) in our bladder and rectum. There are people who have learnt conscious control over Some other autonomic functions; those who are adept at advanced meditation and yoga techniques can voluntarily lower their heartbeat.
And with this, I’ve come to the end of my interesting series of posts on Nervous system. I hope you enjoyed it.
Thank you all for coming. | https://stemgeeks.net/hive-196387/@loveforlove/nervous-systems-parkinson-s-disease-critical-windo-1592277949 |
Part Four: Melody and Harmony
Find the perfect hook as you try out different approaches to creating melodies for your new song.
Jump to section:
Your First Assignment
- After watching the video, reflect on different melodies from songs that you already know.
- What is an example of a melody that you connect with?
- How would you describe the feeling or mood that this melody creates?
- What about the melody specifically creates this mood or feeling?
- Login to Soundtrap and open up your original track. Create a new melody for your track and make sure it works well with the notes from the bassline you created in the last session.
- Review this list of key music production terminology that Charles introduced in the video.
- melody: a sequence of single notes that is musically satisfying
- ear candy: upbeat melodies and catchy choruses that you can’t get out of your head
- harmony: the combination of musical notes sounding simultaneously to produce chords and chord progressions
- triad: a set of three notes played simultaneously
- pentatonic scale: a musical scale with five notes
Your Second Assignment
Listen to the ten chords that Charles plays and write down an emotion you feel for each chord:
1. 6.
2. 7.
3. 8.
4. 9.
5. 10.
Your Third Assignment
- Login to SoundTrap and open up one of your drum and bass loops from the previous lesson.
- Add a chord progression or melody to your loop.
- Once you have added both a melody and harmony to your original song, challenge yourself by creating a new four bar loop starting with chords and melody, and then add other elements, including drums, bass, and vocals.
Explore Other Parts of Digital Music Production
Digital Music Production | Part One: Loops
Explore the origins of modern digital music production and learn how to create loops that will form the basis of your original music.
Digital Music Production | Part Two: Drums
Groove along with Charles as he breaks down the parts of the drum set and uses a digital sequencer to create new drumbeats.
Digital Music Production | Part Three: Bass
Establish the foundation of your original piece of music with a bassline that connects with the drums and sets up the harmonic space for your new tracks.
Digital Music Production| Part Five: Arrangement
Put all of your new digital loops together into a structure that tells a compelling story and provides surprises for your listeners. | https://www.carnegiehall.org/Explore/Learn/Young-Musicians/Digital-Music-Production/Melody-and-Harmony |
· First Response Time is more important than your overall response/reply time because it is an acknowledgment to the customer that their issue is being looked into. It indicates how quickly our team is addressing new support cases, and help us see whether we have enough team members to deal with the volume of the customer´s requests.
Low power sensors generally have faster response times so they will be more susceptible to fluctuating readings when used with a pulsed laser. The position measurement function uses the same thermopile detector as the power reading. If the laser frequency is high enough for the sensor to provide a stable power reading without fluctuations then the position measurement function will perform ...
· Thermocouple Response Time. Time constants calculated for air at room temperature and atmospheric pressure moving with velocity of 65 feet per second for thermocouples shown in Figures #1 and #2. For beaded-type and ungrounded junctions (Figures #3 or #4), multiply time constants by Time constant of thermocouple made with exposed butt ...
• The model has a settling time of ~ 4 , as shown in Section 5. 2. • Because the steady-state gain is one, no offset occurs for set-point changes. • By substituting (12-4) into (12-3b) and solving for Gc, the controller design equation becomes: τc 11 (12-5) c τ c G G s = • The term provides integral control action and thus eliminates offset. • Design parameter provides a convenient
Using the linearity and time reversal properties of the Fourier transform we have X 1(jw) = X 0(jw) + X 0( jw) = 2 2e 1cos(w) 2we 1sin(w) 1 + w2 (b)We know that x 2(t) = x 0(t) x 0( t): Using the linearity and time reversal properties of the Fourier transform we have X 2(jw) = X 0(jw) X 0( jw) = j[2w+ 2e 1sin(w) + 2we 1cos(w) 1 + w2] Problem 2 continued on next page... 4. Aly El Gamal ECE 301 ...
Figure 2: Response of a flrst-order homogeneous equation ¿y_ + y(t) = 0. The efiect of the system time constant ¿ is shown for stable systems (¿ > 0) and unstable systems (¿ < 0). A physical interpretation of the time constant ¿ may be found from the initial condition response of any output variable y(t). If ¿ > 0, the response of any system variable is an exponential decay from the ...
The Wheatstone Bridge diamond shaped circuit who’s concept was developed by Charles Wheatstone can be used to accurately measure unknown resistance values, or as a means of calibrating measuring instruments, voltmeters, ammeters, etc, by the use of a variable resistance and a simple mathematical formula.. Although today digital multimeters provide the simplest way to measure a resistance.
Arduino - Temperature Sensor. The Temperature Sensor LM35 series are precision integrated-circuit temperature devices with an output voltage linearly proportional to the Centigrade temperature. The LM35 device has an advantage over linear temperature sensors calibrated in Kelvin, as the user is not required to subtract a large constant voltage ...
The time required for the detector to respond to an optical input. The response time is related to the bandwidth of the detector by BW = /tr where tris the rise time of the device. The rise time is the time it takes for the detector to rise to a value equal to % of its final steady-state reading. Response time Junction capcacitance 1/RC
2) and a response variable y can be understood as a two-dimensional surface in space. The shape of this surface depends on the structure of the model. The observations are points in space and the surface is “fitted” to best approximate the observations. Example: The simplest multiple regression model for two predictor variables is y = β 0 +β 1 x 1 +β 2 x 2 + The surface that ...
Quantum detectors, another type of sensor, have a higher sensitivity and a lower response time than thermal detectors but require a strong cooling capability (eg, miniature Stirling-type coolers in cameras can achieve temperatures around 70 K). They are made from materials such as InSb, HgCdTe, and GaAs/AIGaAs whose sensitivity depends on photon absorption and, therefore are more expensive as ...
10 4 Lumped analysis is OK. k hL Bi c Bi << , therefore, the lumped approximation is an excellent approximation. The time required for the junction to reach T = 199°C is t s T t T T T b t VC hA b e T T T t T i p bt i ln 1 Transient Conduction in Large Plane Walls, Long Cylinders, and Spheres
Sensory Analysis Section 4 Dr. Bruce W. Zoecklein 4 Table 1. Outline of Sensory Difference and Preference Tests 1 Indicates the minimum number of tasters required for testing to achieve a statistically significant result (p≤).2 Figures denote minimum number of correct responses required out of the total number of responses to conclude the wines are significantly different (p≤) from ...
· The resistance of the LDR sensor changes with the change in intensity of light falling on LDR. This sensor output is given to IC 555 timer connected in bistable mode. The o/p of the IC 555 timer is used to control the prompting of load through a TRIAC. Hence, this circuit switches on the load in the sunset and switches off the load in the sun ...
Thermopile arrays having less than 5,130 elements; a. Non‑“space‑qualified” “focal plane arrays” having all of the following: 1. Individual elements with a peak response within the wavelength range exceeding 900 nm but not exceeding 1,050 nm; and. 2. Any of the following: a. A response “time constant” of less than ns; or. b.
The O2 sensor in your vehicle monitors the exhaust gases coming out of the engine. It determines the amount of oxygen in the gases, which indicates how well the fuel is being burned. The O2 sensor then sends information to the electronic control unit, or ECU, the vehicle's computer. The ECU then adjusts the air-to-fuel ratio to optimize the fuel combustion. Sometimes an O2 sensor fails, and ...
Thermocouples are common temperature sensors used in a wide variety of commercial and industrial applications. While slightly less accurate than resistance temperature detectors (RTDs), thermocouples cover a wide temperature range, are self-powered, and have a fast response time. Their simple construction make them inexpensive and durable. Because of the small sensor voltage and low noise ... | https://bezprocent.pl/30230_which_of_the_following_sensor_have_a_large_response_time_points_thermopile_pyroelectric_sensor_lead_sulphide_sensor_silicon.html |
The 8 queen's puzzle is a well-known puzzle involving placing 8 queens on a chessboard so that no queen attacks another queen. It is more or less obvious that 8 is the maximum number of queens that can populate a chessboard so that no queens are attacking each other.
But what if we use knights instead? How many knights can we maximally place on a chessboard while still satisfying the stipulation of no piece attacking another piece? So far the maximum I've achieved is 24, attained by placing a cluster of 4 knights in the center, and on every square on the edge of the board except for c1, a3, a6, c8, f8, h6, h3, and f1. I cannot see how this number is improved, although I haven't proven that 24 is the theoretical maximum yet.
Additionally, what happens if we use bishops? The maximum I've achieved is 14 at the moment, placing as many as possible along the edges. But this one I haven't really tried to optimize very much. | https://chess.stackexchange.com/questions/32104/maximum-number-of-knights-populating-a-chess-board-so-that-no-knights-are-attack |
Keywords : Hardness, Henna, Physical properties, Polymethylmethacrylate, Surface roughness.
Citation Information : Nawasrah A, Gad MM, Zayat ME. Effect of Henna Addition on the Surface Roughness and Hardness of Polymethylmethacrylate Denture Base Material: An in vitro Study. J Contemp Dent Pract 2018; 19 (6):732-738.
DOI: 10.5005/jp-journals-10024-2327
License: CC BY-NC 3.0
Published Online: 01-06-2018
Copyright Statement: Copyright © 2018; The Author(s).
Aim: This study aimed to evaluate the effect of the addition of various henna—which can have antifungal properties—on the surface roughness and hardness of polymethylmethacrylate (PMMA) denture base material. Materials and methods: A total of 99 rectangular-shaped (10 × 20 × 3 mm3) specimens were prepared from heat-cured acrylic resin and divided into one control group without the addition of henna and five test groups, which were prepared by adding Yamani henna powder to polymer at concentrations of 1, 2.5, 5, 7.5, and 10 wt%. The polymer was added to the monomer, mixed, packed, and processed using the conventional water bath method. After processing, specimens were finished and polished, then kept in distilled water for 48 ± 2 hours. A profilometer and Vickers hardness tester were used to measure surface roughness and hardness respectively. Statistical data analysis was conducted via Statistical Package for the Social Sciences (SPSS) version 20.0 (IBM, USA). The independent sample t-test was used and p ≤0.05 was considered statistically significant. Results: The addition of henna at varying concentrations significantly increased the surface roughness values (p ≤ 0.01) while decreasing hardness (p ≤ 0.0001). The most favorable addition value was 1% henna between all henna groups. Conclusion: The addition of henna to the acrylic resin may negatively affect the surface properties of PMMA acrylic denture base. Clinical significance: Antimicrobial denture with minimum deterioration effects on its physical properties could be achieved with henna addition to denture base material in low concentration. However, 1% henna showed the best results between the henna groups as regards roughness and hardness values. | https://www.thejcdp.com/abstractArticleContentBrowse?paginateValue=12022&abstractArticle=abstractArticle&jpdAbstractArticle=&documentType=Article&jCode=JCDP&jid=19&art_volume=19&art_issue=6&art_page=&numberResult=10&id=12021&sortType= |
Greetings all. Every child with a rough grasp of what life was like in Late Jurassic North America probably knows the Morrison Formation’s main characters. If such a child were to be asked to name the meat-eaters from that formation, the name Ornitholestes would definitely pop up, likely somewhere around third or fourth place.
Ornitholestes was a 6-foot long coelurosaurid theropod dinosaur that lived in western North America during the late Jurassic Period, 155-145 MYA. It is commonly depicted scampering about in the forest, or along the edge of the forest, or sneakily hiding in the shadows out of sight of the larger predators. With the likes of Allosaurus and Torvosaurus stomping around, it’s easy to see why paleo-artists have relegated little Ornitholestes to a bit-part on the Jurassic stage.
But I like to think that Ornitholestes‘ part was much bigger in the never-ending drama of Mesozoic life. Let’s look at its body. I’ve already stated that it was 6 feet long and was therefore about 2 feet tall – large enough to bite you on the knee. It likely weighed a hundred pounds or a smidge less than that – certainly not more. Its skull is worth looking at. Contrary to what has been commonly portrayed, it DID NOT have a little Ceratosaurus-like crest on the end of its nose. That mistake was made when a dislocated bone was mis-identified as a nasal crest. The skull was thin and deep, like a battle axe, and based upon its structure and that of its neck, it likely had a very strong bite. The teeth are small, but they are rather thick in cross-section. A powerful bite and thick teeth? This makes Ornitholestes sound like a precursor to the tyrannosaurs, and no wonder, because the tyrannosaurs are, in fact, highly-evolved coelurosaurs – the same group that Ornitholestes belonged to. The eye sockets on this baby were huge, so it is likely that Ornitholestes was a nocturnal hunter. As for its body, it was a bit on the muscular stocky side, so it was physically strong. It was equipped with long arms ending in three hook-like claws on each hand, and it had a long tail. We can also be fairly sure that Ornitholestes had a coat of thin whispy fur-like feathers on its body since other coelurosaurids that were more primitive and more advanced that Ornitholestes had feathers.
So what can we determine? It was strong for its size, its jaws could crack through eggshells and small bones, it could run, and it could grapple. In short, Ornitholestes was the hyena of the Jurassic savannah.
Hyenas are nothing to laugh at (I’m sorry, that was bad). Hyenas have a reputation for being scavengers, likely because they are commonly seen picking at the leftovers of the lions’ dinner, and because their jaws are the strongest jaws pound-for-pound of any meat-eating animal on the African plains – good for cracking through thick bones of carcasses. But in reality, hyenas are effective hunters as well. They are pack hunters, like lions or wolves, and it’s not unusual to see a gaggle of them, panting and bare-teethed, running down a zebra or a wildebeest.
Was Ornitholestes the same way? Unfortunately, fossils rarely provide evidence for animal behavior. The fact that Ornitholestes fossils are so rare doesn’t help matters. But I dare say that these carnivorous critters were a serious threat to dinosaur mothers who had eggs to protect, they likely did significant damage to hatchlings, they preyed upon smaller animals like thick-boned mammals, and assuredly were seen scavenging carcasses left by other larger meat-eating dinosaurs.
A while back, I drew a picture of Ornitholestes and posted it to this blog. However, it was an “old school” picture portraying Ornitholestes covered in scales. I have recently made an updated version, and I’m posting that image below.
In addition to the feathers, I’ve also slightly altered the shape of the skull to be a little more accurate. I always try to improve my work, and I dare say that a few years from now after my skills have improved further, I’ll make a drawing of this guy that’s even better than the one you see here.
Keep your pencils sharp, people. | https://dinosaursandbarbarians.com/2016/11/28/ornitholestes-with-feathers/ |
Vitamin D is responsible for maintaining normal blood levels of calcium and phosphorus and for supporting bones health. It works with other vitamins, minerals and hormones to promote bone mineralisation. Additionally, research shows that vitamin D may help support a healthy immune system.
The primary source of Vitamin D for both children and adult is from sun exposure. Vitamin D is naturally produced in the skin following exposure to sunlight.
In contrast, very few foods naturally contain vitamin. As a result, a varying percentage of people are vitamin D deficient at any time, with this percentage being higher in the winter, among elderly or obese, in sun-deprived individuals and in dark skinned people.
- Supports bone health
- Helps maintain the immune system
- May help improve mood
Directions
Take 2 tablets daily with a meal
Ingredients
Per 2 tablets
Vitamin D3 100mcg
Other ingredients: Dicalcium phosphate, cellulose, vegetable stearate acid, vegetable magnesium stearate
General Safety Information
- Always read the label. Use only as directed.
- Nutritional supplements may only be of assistance if the dietary intake is inadequate.
- Supplements should not be used to replace a healthy balanced diet and adequate physical exercise.
- Keep all medications and supplements out of reach of children.
- If symptoms persist see your healthcare professional.
Customer Reviews
Payment & Security
Your payment information is processed securely. We do not store credit card details nor have access to your credit card information. | https://www.netpharmacy.co.nz/products/natures-sunshine-vitamin-d3 |
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