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Functional characterization of Class II 5-enopyruvylshikimate-3-phosphate synthase from Halothermothrix orenii H168 in Escherichia coli and transgenic Arabidopsis

文献类型： 外文期刊

作者： Tian, Yong-Sheng 1 ; Xu, Jing 1 ; Xiong, Ai-Sheng 1 ; Zhao, Wei 1 ; Gao, Feng 1 ; Fu, Xiao-Yan 1 ; Peng, Ri-He 1 ; Yao, Q 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai Key Lab Agr Genet & Breeding, Shanghai, Peoples R China

关键词： 5-Enopyruvylshikimate-3-phosphate synthase;Enzyme kinetic values;Halothermothrix orenii;Transgenic Arabidopsis

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

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收录情况： SCI

摘要： Although a large number of AroA enzymes (5-enopyruvylshikimate-3-phosphate synthase [EPSPS]) have been identified, cloned and tested for glyphosate resistance, only AroA variants derived from Agrobacterium tumefaciens strain CP4 have been successfully used commercially. We have now used a polymerase chain reaction (PCR)-based two-step DNA synthesis (PTDS) method to synthesize an aroA gene (aroA _(H. orenii)) from Halothermothrix orenii H168 encoding a new EPSPS similar to AroA _(A. tumefaciens CP4). AroA _(H. orenii) was then expressed in Escherichia coli and key kinetic values of the purified enzyme were determined. Kinetic analysis of AroA _(H. orenii) indicated that the full-length enzyme exhibited increased tolerance to glyphosate compared with E. coli AroA _(E. coli) while retaining a high affinity for the substrate phosphoenolpyruvate. Transgenic Arabidopsis plants containing aroA H. orenii were resistant to 15 mM glyphosate. Site-directed mutagenesis showed that residues Thr355Ser affected the affinity of AroA _(H. orenii) for glyphosate, providing further evidence that specific amino acid residues are responsible for differences in enzymatic behavior among different AroA enzymes.

  • 相关文献

[1]Functional Characterization of 5-Enopyruvylshikimate-3-Phosphate Synthase from Alkaliphilus metalliredigens in Transgenic Arabidopsis. Xing, Xiao-Juan,Sun, Sheng,Xing, Xiao-Juan,Tian, Yong-Sheng,Peng, Ri-He,Xu, Jing,Zhao, Wei,Yao, Quan-Hong.

[2]Identification of a new gene encoding 5-enolpyruvylshikimate-3-phosphate synthase using genomic library construction strategy. Zhou, Chang-Yan,Cai, Run,Wu, Ai-Zhong,Zhou, Chang-Yan,Tian, Yong-Sheng,Xu, Zhi-Sheng,Zhao, Wei,Chen, Chen,Bao, Wen-Hua,Bian, Lin,Wu, Ai-Zhong.

[3]Phytoremediation of triphenylmethane dyes by overexpressing a Citrobacter sp triphenylmethane reductase in transgenic Arabidopsis. Fu, Xiao-Yan,Zhao, Wei,Xiong, Ai-Sheng,Tian, Yong-Sheng,Zhu, Bo,Peng, Ri-He,Yao, Quan-Hong. 2013

[4]Phytoremediation of 2,4,6-trinitrotoluene by Arabidopsis plants expressing a NAD(P)H-flavin nitroreductase from Enterobacter cloacae. You, Shuang-Hong,Zhu, Bo,Han, Hong-Juan,Wang, Bo,Peng, Ri-He,Yao, Quan-Hong.

[5]Functional Characterization of aroA from Rhizobium leguminosarum with Significant Glyphosate Tolerance in Transgenic Arabidopsis. Han, Jing,Tian, Yong-Sheng,Xu, Jing,Wang, Li-Juan,Wang, Bo,Peng, Ri-He,Yao, Quan-Hong.

[6]Enhancement of naphthalene tolerance in transgenic Arabidopsis plants overexpressing the ferredoxin-like protein (ADI1) from rice. Fu, Xiao-Yan,Zhu, Bo,Han, Hong-Juan,Zhao, Wei,Tian, Yong-Sheng,Peng, Ri-He,Yao, Quan-Hong.

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[8]Novel Glyphosate-Resistant aroA Gene from Paracoccus denitrificans in Transgenic Arabidopsis. Tian, Y. -S.,Xiong, A. -S.,Tian, Y. -S.,Yao, Q. -H.,Peng, R. -H.,Xu, J.,Tian, Y. -S.,Xing, X. -J..

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