title: Highly efficient synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate and its derivatives by a robust NADH-dependent reductase from E. coli CCZU-K14.
abstract: An NADH-dependent reductase (CmCR) from Candida magnoliae was discovered by genome mining for carbonyl reductases. After CmCR was overexpressed in Escherichia coli BL21, a robust reductase-producing strain, recombinant E. coli CCZU-K14, was employed for the efficient synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE) from the reduction of ethyl 4-chloro-3-oxobutanoate (COBE). After the optimization, the optimum reaction conditions were obtained. Notably, E. coli CCZU-K14 had broad substrate specificity in reducing both aliphatic and aromatic substrates, and excellent enantioselectivity of CCZU-K14 was observed for most of the tested substrates, resulting in chiral alcohols of over 99.9% ee. Moreover, COBE at a high concentration of (3000mM) could be asymmetrically reduced to (S)-CHBE in the high yield (>99.0%) and high enantiometric excess value (>99.9% ee) after 14h. Significantly, E. coli CCZU-K14 shows high potential in the industrial production of (S)-CHBE and its derivatives (>99.9% ee).
authors.affiliation: Laboratory of Biocatalysis and Bioprocessing, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou 213164, PR China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, PR China. Electronic address: heyucai2001@163.com.
authors.forename: Yu-Cai
authors.initials: YC
authors.lastname: He
authors.affiliation: Laboratory of Biocatalysis and Bioprocessing, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou 213164, PR China.
authors.forename: Zhi-Cheng
authors.initials: ZC
authors.lastname: Tao
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, PR China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Laboratory of Biocatalysis and Bioprocessing, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou 213164, PR China.
authors.forename: Zhen-Xing
authors.initials: ZX
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, PR China.
authors.forename: Jian-He
authors.initials: JH
authors.lastname: Xu
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D002087:Butyrates / Q000138:chemical synthesis*; D002175:Candida / Q000201:enzymology*; D004926:Escherichia coli; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D002087:Butyrates; D011994:Recombinant Proteins; C121244:ethyl 4-chloro-3-hydroxybutanoate; D000429:Alcohol Oxidoreductases
keywords: Asymmetric reduction; Biotransformation; Ethyl (S)-4-chloro-3-hydroxybutanoate; Ethyl 4-chloro-3-oxobutanoate; Recombinant E. coli CCZU-K14