数字农科院2.0

A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution

文献类型： 外文期刊

作者： Yuan, Yuan;Zhou, Zhengfu;Zhan, Yuhua;Ke, Xiubin;Yan, Yongliang;Lin, Min;Li, Pengcheng;Jiang, Shijie;Wang, Jin;Lu, Wei

作者机构：

关键词： 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS);adaptive laboratory evolution (ALE);mutation;glyphosate-resistance;enzyme kinetics

期刊名称： APPLIED SCIENCES-BASEL

ISSN：

年卷期： 2022 年 12 卷 11 期

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摘要： EPSP synthase is the target enzyme of glyphosate herbicides. Due to the extensive use of glyphosate, it is very important to obtain EPSPS genes with high glyphosate resistance for the development of transgenic crops. GR79-EPSPS is a class I EPSP synthase with certain glyphosate resistance isolated from glyphosate-contaminated soil. After more than 1000 generations, a Y40I substitution was identified, and the enzyme had a nearly 1.8-fold decrease in Km [PEP] and a 1.7-fold increase in Ki[glyphosate] compared to the wild-type enzyme. Enzyme dynamics and molecular dynamics analysis showed that the substitution was near the hinge region of EPSPS, and the affinity of glyphosate binding to amino acid residues of the active site decreased due to Y40I substitution, resulting in an increase in glyphosate resistance. These results provide more evidence for the combination of directed evolution and rational design of protein engineering.

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