A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution
EPSP synthase is the target enzyme of glyphosate herbicides. Due to the extensive use of glyphosate, it is very important to obtain <i>EPSPS</i> genes with high glyphosate resistance for the development of transgenic crops. GR79-EPSPS is a class I EPSP synthase with certain glyphosate re...
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MDPI AG
2022-06-01
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| author | Yuan Yuan Zhengfu Zhou Yuhua Zhan Xiubin Ke Yongliang Yan Min Lin Pengcheng Li Shijie Jiang Jin Wang Wei Lu |
| author_facet | Yuan Yuan Zhengfu Zhou Yuhua Zhan Xiubin Ke Yongliang Yan Min Lin Pengcheng Li Shijie Jiang Jin Wang Wei Lu |
| author_sort | Yuan Yuan |
| collection | DOAJ |
| description | EPSP synthase is the target enzyme of glyphosate herbicides. Due to the extensive use of glyphosate, it is very important to obtain <i>EPSPS</i> 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 <i>K</i>m [PEP] and a 1.7-fold increase in <i>K</i>i[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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| spelling | doaj.art-138147d869174c9c8d873f48b4b8214b2023-11-23T13:46:47ZengMDPI AGApplied Sciences2076-34172022-06-011211572310.3390/app12115723A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory EvolutionYuan Yuan0Zhengfu Zhou1Yuhua Zhan2Xiubin Ke3Yongliang Yan4Min Lin5Pengcheng Li6Shijie Jiang7Jin Wang8Wei Lu9School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaSchool of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, ChinaSchool of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaEPSP synthase is the target enzyme of glyphosate herbicides. Due to the extensive use of glyphosate, it is very important to obtain <i>EPSPS</i> 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 <i>K</i>m [PEP] and a 1.7-fold increase in <i>K</i>i[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.https://www.mdpi.com/2076-3417/12/11/57235-enolpyruvylshikimate-3-phosphate synthase (EPSPS)adaptive laboratory evolution (ALE)mutationglyphosate-resistanceenzyme kinetics |
| spellingShingle | Yuan Yuan Zhengfu Zhou Yuhua Zhan Xiubin Ke Yongliang Yan Min Lin Pengcheng Li Shijie Jiang Jin Wang Wei Lu A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution Applied Sciences 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) adaptive laboratory evolution (ALE) mutation glyphosate-resistance enzyme kinetics |
| title | A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution |
| title_full | A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution |
| title_fullStr | A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution |
| title_full_unstemmed | A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution |
| title_short | A Highly Glyphosate-Resistant EPSPS Mutant from Laboratory Evolution |
| title_sort | highly glyphosate resistant epsps mutant from laboratory evolution |
| topic | 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) adaptive laboratory evolution (ALE) mutation glyphosate-resistance enzyme kinetics |
| url | https://www.mdpi.com/2076-3417/12/11/5723 |
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