EPSPS Enzyme Mutant for Glyphosate-Resistant Crops
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Solution Overview
Problem
Existing glyphosate herbicides efficiently control weeds but non-selectively damage crops, limiting the use of transgenic crops with glyphosate resistance and necessitating manual weeding and high weeding costs.
Innovation Solution
Development of a glyphosate-resistant EPSPS enzyme mutant with specific amino acid mutations at sites 131, 172, 177, 274, 313, and 403, enabling plants to express a glyphosate-resistant EPSPS enzyme, which is integrated into plants through expression cassettes and vectors, enhancing crop tolerance to glyphosate.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If glyphosate is used to control weeds, then weed control efficiency is improved, but crops are damaged due to non-selective action
Solution Approach 1:
The patent introduces specific amino acid mutations at six key sites (131, 172, 177, 274, 313, 403) in the EPSPS enzyme structure. These localized changes create a glyphosate-resistant enzyme variant that maintains catalytic function while resisting herbicide inhibition, allowing crops to tolerate glyphosate application for weed control
Solution Approach 2:
The patent modifies the biochemical parameters of the EPSPS enzyme through amino acid substitution. The mutant enzyme has altered binding characteristics that prevent glyphosate from effectively inhibiting the shikimic acid pathway, while maintaining sufficient enzymatic activity for aromatic amino acid synthesis in the presence of glyphosate
2Productivity
If transgenic crops with glyphosate resistance are cultivated, then manual weeding burden is reduced and crop yields increase, but transgenic technology acceptance is limited
Solution Approach 1:
The patent creates a plant-derived copy of the glyphosate-resistant EPSPS enzyme (CP4-EPSPS) that can be expressed in crops. This approach uses a naturally occurring resistant enzyme from a plant-associated bacterium, which is then genetically engineered into crops to confer glyphosate resistance, reducing manual weeding and increasing yields
3Reliability
If CP4-EPSPS gene is introduced into crops, then glyphosate resistance is achieved, but resistance level is insufficient compared to novel mutants
Solution Approach 1:
The patent combines multiple amino acid mutations at six different sites in the EPSPS enzyme to create a composite mutant with enhanced glyphosate resistance. This multi-site mutation strategy produces a synergistic effect that provides superior resistance compared to single mutations or the original CP4-EPSPS gene
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 glyphosate-resistant EPSPS enzyme mutant improves crop yields and reduces weeding costs by conferring higher tolerance to glyphosate than existing CP4-EPSPS genes, facilitating the cultivation of glyphosate-resistant crops like rice and soybean.
Implementation Method 1
EPSPS is an enzyme involved in a shikimic acid biosynthetic pathway, and involved in synthesis of aromatic amino acids
Implementation Method 2
an action mechanism thereof primarily involves competitive inhibition of EPSPS activity in the shikimic acid pathway, thus preventing affected plants from continuously synthesizing essential aromatic amino acids
Data Source
AI summary
Provided are an EPSPS enzyme mutant, a nucleic acid molecule, an expression cassette, an expression vector, a recombinant bacterium or a recombinant cell and use thereof. The EPSPS enzyme mutant can effectively solve the problem of non-selectable damage to crops caused by herbicides such as glyphosate, and provides a new route for cultivating glyphosate-resistant crop varieties. Moreover, the EPSPS enzyme mutant also has relatively high practicability and promotional value, can improve a weed control effect, reduce weeding costs, and elevate crop yields in agricultural production. Besides, it further provides an expression cassette expressing the EPSPS enzyme mutant, where the expression cassette contains an EPSPS mutant gene conferring glyphosate resistance on a variety of plants such as soybean and rice.