EPSP Synthase Mutations for Glyphosate Resistance
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Solution Overview
Problem
Current methods for conferring herbicide resistance in plants and bacteria are limited by the toxicity of glyphosate to EPSP synthase enzymes, necessitating the development of genes encoding glyphosate-resistant variants to enhance tolerance and biosynthesis pathways.
Innovation Solution
The use of nucleic acid molecules encoding herbicide-resistant or tolerant polypeptides, specifically the sequences provided, which are used to transform organisms to confer resistance, including plasmids deposited in the NRRL collection, allowing for the expression of glyphosate-resistant EPSP synthase enzymes that maintain catalytic activity in the presence of glyphosate.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If glyphosate is used as a herbicide, then plant cells are killed by inhibiting EPSP synthase, but bacterial cells with wild-type EPSP synthase are also toxic to
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (mutations) at positions 42 (T42M), 230 (Q230K), and/or 102 (TIPS) in the EPSP synthase protein sequence. These parameter changes in the protein's amino acid composition alter the enzyme's binding affinity for glyphosate, reducing toxicity while maintaining catalytic activity, thereby conferring herbicide resistance on transformed plants and bacteria
2Adaptability or versatility
If mutant EPSP synthase genes are introduced to confer herbicide resistance, then glyphosate tolerance is achieved, but the complexity of genetic transformation increases
Solution Approach 1:
The patent extracts and isolates specific functional regions and key mutation sites from the EPSP synthase gene. By identifying and extracting only the critical mutation positions (42, 230, and 102) rather than the entire gene sequence, the complexity of genetic transformation is reduced while maintaining herbicide resistance functionality
Solution Approach 2:
The patent segments the EPSP synthase gene into distinct functional regions and mutation sites. This segmentation allows for targeted modification at specific positions (T42M, Q230K, TIPS) independently, simplifying the genetic transformation process and enabling modular approach to creating herbicide-resistant organisms
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The described approach enables plants and bacteria to exhibit increased tolerance to glyphosate, allowing them to survive and function in environments with higher glyphosate concentrations, thereby enhancing agricultural applications and resistance to herbicide toxicity.
Implementation Method 1
Glyphosate inhibits the enzyme that converts phosphoenolpyruvic acid (PEP) and 3-phosphoshikimic acid to 5-enolpyruvyl-3-phosphoshikimic acid. Inhibition of this enzyme (5-enolpyruvylshikimate-3-phosphate synthase; referred to herein as 'EPSP synthase') kills plant cells by shutting down the shikimate pathway
Implementation Method 2
These enzymes contain amino acid substitutions in their active sites that prevent the binding of glyphosate without affecting binding by PEP or S3P
Data Source
AI summary
Compositions and methods for conferring herbicide resistance or tolerance to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a polypeptide that confers resistance or tolerance to glyphosate herbicides are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated nucleic acid molecules corresponding to glyphosate resistant nucleic acid sequences are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in SEQ ID NOS:2, 4, 6, 8, 10, 12, or 14 or the nucleotide sequence set forth in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 28, 29, 30, 31, 32, 33, or 34.