title: Discovery of a reductase-producing strain recombinant E. coli CCZU-A13 using colorimetric screening and its whole cell-catalyzed biosynthesis of ethyl (R)-4-chloro-3-hydroxybutanoate.
abstract: An NADH-dependent reductase (SsCR) was discovered by genome data mining. After SsCR was overexpressed in E. coli BL21, recombinant E. coli CCZU-A13 with high reductase activity and excellent stereoselectivity for the reduction of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl (R)-4-chloro-3-hydroxybutanoate ((R)-CHBE) was screened using one high-throughput colorimetric screening strategy. After the reaction optimization, a highly stereoselective bioreduction of COBE into (R)-CHBE (>99% ee) with the resting cells of E. coli CCZU-A13 was successfully demonstrated in n-butyl acetate-water (10:90, v/v) biphasic system. Biotransformation of 600mM COBE for 8h in the biphasic system, (R)-CHBE (>99% ee) could be obtained in the high yield of 100%. Moreover, the broad substrate specificity in the reduction of aliphatic and aromatic carbonyl compounds was also found. Significantly, E. coli CCZU-A13 shows high potential in the industrial production of (R)-CHBE (>99% ee) and its derivatives.
authors.affiliation: Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, Changzhou University, Changzhou, PR China. Electronic address: heyucai2001@163.com.
authors.forename: Yu-Cai
authors.initials: YC
authors.lastname: He
authors.affiliation: Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, Changzhou University, Changzhou, PR China.
authors.forename: Dan-Ping
authors.initials: DP
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, Changzhou University, Changzhou, PR China.
authors.forename: Zhi-Cheng
authors.initials: ZC
authors.lastname: Tao
authors.affiliation: Department of Bioengineering, Rice University, Houston, TX, USA.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, Changzhou University, Changzhou, PR China.
authors.forename: Zhen-Xing
authors.initials: ZX
authors.lastname: Yang
mesh_terms: D000090:Acetoacetates / Q000302:isolation & purification / Q000378:metabolism*; D002384:Catalysis; D003124:Colorimetry / Q000379:methods*; D004926:Escherichia coli / Q000145:classification* / Q000502:physiology*; D024861:Genetic Enhancement / Q000379:methods*; D009243:NAD / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D000090:Acetoacetates; D011994:Recombinant Proteins; D009243:NAD; C111982:ethyl 4-chloro-3-oxobutanoate
keywords: Asymmetric bioreduction; Biphasic system; Ethyl (R)-4-chloro-3-hydroxybutanoate; Ethyl 4-chloro-3-oxobutanoate; Recombinant E. coli CCZU-A13