文献类型：期刊文献

英文题名：Efficient Biocatalytic Synthesis of (R)-2-Chloro-1-(3,4-difluorophenyl)ethanol by the Short-Chain Dehydrogenase PpKR8 from Paraburkholderia phymatum STM815

作者：He, Ling[1];Ye, Wenjie[1];Xie, Youyu[1];Liu, Qinghai[1];Wang, Hualei[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2022

卷号：26

期号：2

起止页码：278

外文期刊名：ORGANIC PROCESS RESEARCH & DEVELOPMENT

收录：;EI(收录号：20220511555581);WOS:【SCI-EXPANDED(收录号:WOS:000758184400006)】；

基金：The authors are grateful for the support provided by the National Natural Science Foundation of China (No. 21776084) and the Fundamental Research Funds for the Central Universities. The authors thank Editage ( www.editage. cn) for English language editing.

语种：英文

外文关键词：(R)-2-chloro-1-(3,4-difluorophenyl)ethanol; alcohol dehydrogenase; asymmetric reduction; coexpression

摘要：Bioreductions catalyzed by ketoreductases play an important role in the synthesis of chiral alcohols. However, the synthesis of (R)-2-chloro-1-(3,4-difluorophenyl)ethanol (24b), an important chiral intermediate for the synthesis of the anticoagulant ticagrelor, poses significant challenges in terms of high substrate concentration requirements that limit its production. In this study, a novel NADH-dependent, short-chain dehydrogenase, PpKR8, from Paraburkholderia phymatum STM815, exhibited excellent enantioselectivity and high activity for the production of (R)-CFPL (24b) through the reduction of 2-chloro-1-(3,4difluorophenyl)ethanone (24a). The coexpression of PpKR8 and glucose dehydrogenase from Bacillus subtilis in Escherichia coli allowed up to 300 g/L (1.57 M) CFPO (24a) to be completely converted into 24b with 99.9% enantiomeric excess and a high space-time yield (728 g/(L day)). Substrate specificity assays demonstrated a broad substrate spectrum for PpKR8, which included 35 alpha/beta-ketoesters, aromatic ketones, and heterocyclic ketones. Moreover, three additional optically pure chiral alcohols that are used as important drug intermediates were synthesized at high substrate concentrations (150-330 g/L), demonstrating the excellent industrial potential of PpKR8-mediated bioreductions.