Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (R)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis

作     者：Bingmei Su Lian Xu Xinqi Xu Lichao Wang Aipeng Li Juan Lin Lidan Ye Hongwei Yu 

作者机构：College of Chemical EngineeringFuzhou UniversityFuzhou 350116China College of Biological Science and EngineeringFuzhou UniversityFuzhou 350116China Institute of BioengineeringCollege of Chemical and Biological EngineeringZhejiang UniversityHangzhou 310027China School of Life SciencesNorthwestern Polytechnical UniversityXi’an 710072China 

出 版 物：《Green Synthesis and Catalysis》 (绿色合成与催化(英文))

年 卷 期：2020年第1卷第2期

页      面：150-159页

学科分类：081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 070303[理学-有机化学] 0703[理学-化学] 

基　　金：This study is financially supported by the National Key Research and Development Program of China(No.2018YFA0901800 to H.Yu and L.Ye) Zhejiang Provincial Natural Science Foundation of China(No.LY18B060001 to L.Ye and No.LZ20B060002 to H.Yu) Natural Science Foundation of China(No.21776244 to H.Yu).We are grateful to the constructive suggestions from academician Fen-Er Chen(Department of Chemistry,Fudan University,Shanghai,China)and the support of Discovery Studio software from Fuzhou University platform 

主　　题：Ethyl(R)-2-hydroxy-4-phenylbutanoate Rational design Enantioselectivity Short-chain dehydrogenase/reductase Per-residue free energy decomposition Sequence conservatism analysis 

摘      要：As an important building block for the synthesis of angiotensin-converting enzyme inhibitors,ethyl(R)-2-hy-droxyl-4-phenylbutanoate[(R)-HPBE]has attracted increasing *** key to industrial biosynthesis of(R)-HPBE is a biocatalyst that efficiently reduces ethyl 2-oxo-4-phenylbutanoate(OPBE)with high *** paper proposed a strategy for identifying key residues involved in enantioselectivity control based on per-residue free energy decomposition and sequence conservatism *** this strategy,4 noncon-servative sites with high energy contribution to binding of OPBE were chosen as engineering targets,generating variant Mu27 with 99%conversion and 98%(R)ee value at substrate loading of up to 500 mmol/*** simu-lations suggested the higher stability and formation probability of Mu27-OPBEproR prereaction state as key rea-sons for the excellent R-enantioselectivity of Mu27 towards *** success in this study provides a viable approach for rational design of alcohol dehydrogenases with high enantioselectivity towards unnatural substrates.