Characterization of acid‑resistant aldo–keto reductases capable of asymmetric synthesis of(R)‑CHBE from Lactobacillus plantarum DSM20174  

作　　者：Jiufeng Zhu Yajun Bai Tai‑Ping Fan Xiaohui Zheng Yujie Cai 

机构地区：[1]The Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi 214122,Jiangsu,China [2]College of Life Sciences,Northwest University,Xi’an 710069,Shanxi,China [3]Department of Pharmacology,University of Cambridge,Cambridge CB21T,UK

出　　处：《Systems Microbiology and Biomanufacturing》2023年第4期634-646,共13页系统微生物学与生物制造（英文）

摘　　要：Ethyl(R)-4-chloro-3-hydroxybutyrate((R)-CHBE),as a chiral intermediate,is widely used in the synthesis of various chiral drugs.In this study,we screened two aldo–keto reductases(LP-AKRs)from the probiotic Lactobacillus plantarum DSM20174,both with a molecular weight of approximately 31 kDa.Both enzymes could reduce 4-chloroacetoacetic acid ethyl ester(COBE)to produce(R)-CHBE with an enantioselectivity value of 99%.When determining the kinetic parameter,the K_(m),K_(cat),and V_(max)of LP-AKR5 and LP-AKR9 were 9.5 mM,7.6 U/mg,3.96 s^(-1)and 8.7 mM,8.59 U/mg,4.47 s^(-1),respectively.Both LP-AKR5 and LP-AKR9 had an optimal reaction pH of 6 and could maintain a high level of stability at pH 6,allowing them to perform well in an acidic environment.LP-AKR5 and LP-AKR9 had optimal reaction temperatures of 30℃and 40℃,respectively.Metal ions had minimal influence on LP-AKR5 and LP-AKR9 enzyme activities.This series of enzymatic properties showed that LP-AKR5 and LP-AKR9 mined from Lactobacillus plantarum DSM20174 could asym-metrically catalyze the synthesis of(R)-CHBE under weakly acidic circumstances,which could maintain product stability and provide a good foundation for industrial production.

关 键 词：Aldo–keto reductases Lactobacillus plantarum CHARACTERIZATION (R)-CHBE 

分 类 号：Q93[生物学—微生物学]