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| Expression of a Novel Epoxide Hydrolase from Phaseolus vulgaris and Its Enantioconvergent Catalytic Performance |
| YE Hui-hua1, HU Die2, LI Chuang2, CHENG Jian-qing3, DENG Chao3, WU Min-chen3 |
1. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China;
2. School of Biotechnology, Jiangnan University, Wuxi 214122, China;
3. Wuxi Medical School, Jiangnan University, Wuxi 214122, China |
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Abstract To excavate a novel epoxide hydrolase from Phaseolus vulgaris (PvEH1) and explore its enantioconvergent catalytic performance, a PvEH1-encoding gene (pveh1) was amplified from the P. vulgaris total RNA by RT-PCR technique. Then, pveh1 was heterologously expressed in E. coli BL21(DE3) mediated by an expression plasmid pET28a(+). Analysis of primary and three-dimensional structures indicated that the identities of PvEH1 with Vigna radiata and Medicago truncatula epoxide hydrolases are 85.7 and 81.1%. Its catalytic triad is Asp101-His299-Asp264, belonging to the α/β-hydrolase superfamily. When the conversion rate of racemic styrene oxide catalyzed by PvEH1 reached 99.1%, the product, (R)-1-phenyl-1,2-ethanediol, was obtained with an enantiomeric purity of 33.6% e.e.p. PvEH1 possesses the opposite regioselectivity towards (S)-SO and (R)-SO with regioselectivity coefficients (αS and βR) of 91.1 and 53.3%. The discovery of PvEH1 and characterization of its enantioconvergence not only increased the number of plant EHs, but also established a foundation for the study of its catalytic mechanism and the directed modification of its regioselectivity.
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Received: 05 April 2016
Published: 25 October 2016
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