化工进展2025，Vol.44Issue(5)：2515-2523,9.DOI:10.16085/j.issn.1000-6613.2024-1851
半理性设计提高短链醇脱氢酶在(S)-1-(4-氟苯基)乙醇合成中的应用
Semi-rational design to enhance short-chain alcohol dehydrogenases in the synthesis of(S)-1-(4-fluorophenyl)ethanol
摘要
Abstract
(S)-1-(4-fluorophenyl)ethanol is an important chiral drug intermediate,which is widely used in the synthesis of various drugs such as Alzheimer's disease therapeutics.Currently,preparation inefficiencies are prevalent in the synthesis of this chiral alcohol using bioasymmetric reduction.In order to establish an enzymatic and efficient synthesis process of(S)-1-(4-fluorophenyl)ethanol,this study screened a laboratory enzyme library containing 323 alcohol dehydrogenases,obtained DpADH,a short-chain alcohol dehydrogenase possessing high stereoselectivity(e.e.value of 99.9%)and relative activity,and further improved the catalytic performance of the enzyme by semi-rational design.The excellent variant M3(N164C/S195W/F157A)with a 30.8-fold increase in catalytic efficiency was successfully obtained using multiple rounds of iterative mutagenesis.To improve the efficiency of coenzyme delivery,a co-expression strain of alcohol dehydrogenase variant M3 with glucose dehydrogenase BsGDH was constructed,which showed 98.5%conversion of 4-fluoroacetophenone(100g/L)within 7h of reaction.Molecular dynamics simulations showed that the variant M3 increased the favourable π-π interactions with the substrate and stabilised the substrate-bound conformation.This study provides an economical and efficient synthetic route for(S)-1-(4-fluorophenyl)ethanol,and provides guidance for the molecular modification and mechanistic elucidation of DpADH,which enhances the potential of the enzyme for industrial applications.关键词
(S)-1-(4-氟苯基)乙醇/半理性设计/醇脱氢酶/不对称还原/共表达Key words
(S)-1-(4-fluorophenyl)ethanol/semi-rational design/alcohol dehydrogenase/asymmetric reduction/co-expression分类
生物科学引用本文复制引用
谢静雯,孟仪方,叶文杰,王华磊,魏东芝..半理性设计提高短链醇脱氢酶在(S)-1-(4-氟苯基)乙醇合成中的应用[J].化工进展,2025,44(5):2515-2523,9.基金项目
国家自然科学基金(22078097). （22078097）
