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Expression of a novel epoxide hydrolase of Aspergillus usamii E001 in Escherichia coli and its performance in resolution of racemic styrene oxide

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The full-length cDNA sequence of Aueh2, a gene encoding an epoxide hydrolase of Aspergillus usamii E001 (abbreviated to AuEH2), was amplified from the total RNA. Synchronously, the complete DNA sequence containing 5′, 3′ flanking regions, eight exons and seven introns was cloned from the genomic DNA. In addition, a cDNA fragment of Aueh2 encoding a 395-aa AuEH2 was expressed in Escherichia coli. The catalytic activity of recombinant AuEH2 (re-AuEH2) was 1.44 U/ml using racemic styrene oxide (SO) as the substrate. The purified re-AuEH2 displayed the maximum activity at pH 7.0 and 35 °C. It was highly stable at a pH range of 5.0–7.5, and at 40 °C or below. Its activity was not obviously influenced by β-mercaptoethanol, EDTA and most of metal ions tested, but was inhibited by Hg2+, Sn2+, Cu2+, Fe3+ and Zn2+. The K m and V max of re-AuEH2 were 5.90 mM and 20.1 U/mg towards (R)-SO, while 7.66 mM and 3.19 U/mg towards (S)-SO. Its enantiomeric ratio (E) for resolution of racemic SO was 24.2 at 10 °C. The experimental result of re-AuEH2 biasing towards (R)-SO was consistent with the analytical one by molecular docking (MD) simulation.

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Acknowledgments

This work was financially supported by the National Nature Science Foundation of China (No. 31271811), the Fundamental Research Funds for the Central Universities of China (JUSRP51412B), and the Postgraduate Innovation Training Project of Jiangsu, China (No. CXZZ13_0757). We are grateful to Prof. Xianzhang Wu (School of Biotechnology, Jiangnan University, China) for providing technical assistance.

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    Authors and Affiliations

    1. Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People’s Republic of China

      Die Hu

    2. China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, 1638 Wolong Road, Nanyang, 473061, Henan, People’s Republic of China

      Cun-Duo Tang

    3. Wuxi Medical School, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People’s Republic of China

      Biao Yang, Jia-Chi Liu, Chao Deng & Min-Chen Wu

    4. School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People’s Republic of China

      Tao Yu

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    1. Die Hu
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    Correspondence to Min-Chen Wu.

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    D. Hu and C.-D. Tang contributed equally to this work.

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    Hu, D., Tang, CD., Yang, B. et al. Expression of a novel epoxide hydrolase of Aspergillus usamii E001 in Escherichia coli and its performance in resolution of racemic styrene oxide. J Ind Microbiol Biotechnol 42, 671–680 (2015). https://doi.org/10.1007/s10295-015-1604-y

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