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Characterization of a class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate from Sinorhizobium fredii

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Abstract

5-Enopyruvylshikimate-3-phosphate synthase (EPSP synthase) is an important enzyme in the shikimate pathway mediating the biosynthesis of aromatic compounds in plants and microorganisms. A novel class II EPSP synthase AroA S. fredii from Sinorhizobium fredii NGR234 was overexpressed in Escherichia coli BL21. It was purified to homogeneity and its catalytic properties were studied. The enzyme exhibited optimum catalytic activity at pH 8.0 and 50 °C. It was stable below 40 °C, and over a broad range of pH 5.0–9.0. The EPSP synthase was increasingly activated by 100 mM of the chlorides of NH4 +, K+, Na+ and Li+. Kinetic analysis of AroA S. fredii suggested that the enzyme exhibited a high glyphosate tolerance and high level of affinity for phosphoenolpyruvate, which indicates the enzyme with a high potential for structural and functional studies and its potential usage for the generation of transgenic crops resistant to the herbicide.

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Acknowledgments

The research was supported by the Key Project Fund of the Shanghai Municipal Committee of Agriculture (No. 2011-1-8), International Scientific and Technological Cooperation (2010DFA62320, 11230705900), the Youth Fund of Shanghai Academy of Agricultural Sciences (2012-16, 2013-16), National Natural Science Foundation (31071486) and the Key Project Fund of Shanghai Minhang Science and Technology Committee (2012MH059).

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Authors and Affiliations

  1. Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, China

    Lijuan Wang, Rihe Peng, Yongsheng Tian, Jing Han, Wei Zhao, Bo Wang, Man Liu & Quanhong Yao

  2. College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China

    Quanhong Yao

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  1. Lijuan Wang
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  2. Rihe Peng
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  3. Yongsheng Tian
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  4. Jing Han
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  5. Wei Zhao
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  6. Bo Wang
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  7. Man Liu
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  8. Quanhong Yao
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Correspondence to Man Liu or Quanhong Yao.

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Wang, L., Peng, R., Tian, Y. et al. Characterization of a class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate from Sinorhizobium fredii . World J Microbiol Biotechnol 30, 2967–2973 (2014). https://doi.org/10.1007/s11274-014-1724-y

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  • DOI: https://doi.org/10.1007/s11274-014-1724-y

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