文献类型：期刊文献

英文题名：Efficient Asymmetric Synthesis of Ethyl (S)-4-Chloro-3-hydroxybutyrate Using Alcohol Dehydrogenase SmADH31 with High Tolerance of Substrate and Product in a Monophasic Aqueous System

作者：Yang, Zeyu[1];Ye, Wenjie[1];Xie, Youyu[1];Liu, Qinghai[1];Chen, Rong[2];Wang, Hualei[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China;[2]Hangzhou Normal Univ, Sch Med, Hangzhou 311121, Peoples R China

年份：2020

卷号：24

期号：6

起止页码：1068

外文期刊名：ORGANIC PROCESS RESEARCH & DEVELOPMENT

收录：;EI(收录号：20202708893933);WOS:【SCI-EXPANDED(收录号:WOS:000543672300015)】；

基金：This work was supported by the National Natural Science Foundation of China (Nos. 21776084 and 21606062) and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：asymmetric synthesis; ethyl (S)-4-chloro-3-hydroxybutanoate; alcohol dehydrogenases; high substrate and product tolerance; monophasic aqueous system

摘要：Bioreductions catalyzed by alcohol dehydrogenases (ADHs) play an important role in the synthesis of chiral alcohols. However, the synthesis of ethyl (S)-4-chloro-3-hydroxybutyrate [(S)-CHBE], an important drug intermediate, has significant challenges concerning high substrate or product inhibition toward ADHs, which complicates its production. Herein, we evaluated a novel ADH, SmADH31, obtained from the Stenotrophomonas maltophilia genome, which can tolerate extremely high concentrations (6 M) of both substrate and product. The coexpression of SmADH31 and glucose dehydrogenase from Bacillus subtilis in Escherichia coli meant that as much as 660 g L-1 (4.0 M) ethyl 4-chloroacetoacetate was completely converted into (S)-CHBE in a monophasic aqueous system with a >99.9% ee value and a high space-time yield (2664 g L-1 d(-1)). Molecular dynamics simulation shed light on the high activity and stereoselectivity of SmADH31. Moreover, five other optically pure chiral alcohols were synthesized at high concentrations (100-462 g L-1) as a result of the broad substrate spectrum of SmADH31. All these compounds act as important drug intermediates, demonstrating the industrial potential of SmADH31-mediated bioreductions.