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发明公开
CN107988308A 一种立体选择性提高的菜豆环氧水解酶突变体及其应用 有权

一种立体选择性提高的菜豆环氧水解酶突变体及其应用
摘要：
本发明公开一种立体选择性提高的菜豆环氧水解酶突变体及其应用，属于酶工程技术领域。本发明通过菜豆来源的环氧水解酶进行单向定点突变，获得对映选择性与对映归一性提高的突变酶W102L。与野生型相比，W102L对外消旋对氯环氧苯乙烷rac-pCSO的E值从2.6提高至25.5，对rac-pCSO的对映选择性提高了9.8倍。使用PvEH1W102L全细胞动力学拆分150mmol·L-1的rac-pCSO，反应4h后，可获得(R)-对氯环氧苯乙烷的产率为45.62％(ees＝96.30％)和(R)-对氯苯基乙二醇产率为50.91％(eep＝90.26％)。
摘要（英）：

The invention discloses a phaseolus vulgaris epoxide hydrolase mutant with improved stereoselectivity and application thereof, and belongs to the technical field of enzyme engineering. According to the invention, unidirectional site-directed mutagenesis is carried out by epoxide hydrolase from phaseolus vulgaris so as to obtain a mutant enzyme W102L with improved enantioselectivity and enantio-polarity. Compared to a wild type, an E value of the W102L for racemized p-chlorobenzenethiol styrene oxide rac-pCSO is increased to 25.5 from 2.6, and enantioselectivity of the W102L for the rac-pCSO isimproved by 9.8 times. 150mmol.L-1 of rac-pCSO is resolved by using PvEH1W102L whole-cell dynamics, and after a reaction is performed by 4h, yield of 45.62% (ees=96.30%) of (R)-p-chlorobenzenethiol styrene oxide and yield of 50.91% (eep=90.26%) of (R)-p-chlorobenzenethiol phenyl ethylene glycol can be obtained.

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