title: Biosynthesis of ethyl (S)-4-chloro-3-hydroxybutanoate by NADH-dependent reductase from E. coli CCZU-Y10 discovered by genome data mining using mannitol as cosubstrate.
abstract: The reductase (PgCR) from recombinant Escherichia coli CCZU-Y10 displayed high reductase activity and excellent stereoselectivity for the reduction of ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE). To efficiently synthesize (S)-CHBE (>99 % enantiomeric excess (ee)), the highly stereoselective bioreduction of COBE into (S)-CHBE with the whole cells of E. coli CCZU-Y10 was successfully demonstrated in a dibutyl phthalate-water biphasic system. The appropriate ratio of the organic phase to water phase was 1:1 (v/v). The optimum reaction temperature, reaction pH, cosubstrate, NAD(+), and cell dosage of the biotransformation of 100 mM COBE in this biphasic system were 30 °C, 7.0, mannitol (2.5 mmol/mmol COBE), 0.1 μmol/(mmol COBE), and 0.1 g (wet weight)/mL, respectively. Moreover, COBE at a high concentration of (1,000 mM) could be asymmetrically reduced to (S)-CHBE in a high yield (99.0 %) and high enantiometric excess value (>99 % ee). Significantly, E. coli CCZU-Y10 shows high potential in the industrial production of (S)-CHBE (>99 % ee).
authors.affiliation: Laboratory of Biocatalysis and Bioprocessing, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, 213164, People's Republic of China, heyucai2001@163.com.
authors.forename: Yu-Cai
authors.initials: YC
authors.lastname: He
authors.forename: Zhen-Xing
authors.initials: ZX
authors.lastname: Yang
authors.forename: Dan-Ping
authors.initials: DP
authors.lastname: Zhang
authors.forename: Zhi-Cheng
authors.initials: ZC
authors.lastname: Tao
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Chen
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.forename: Jian-He
authors.initials: JH
authors.lastname: Xu
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
authors.forename: Rui-Jie
authors.initials: RJ
authors.lastname: Chen
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Ma
mesh_terms: D000090:Acetoacetates / Q000737:chemistry / Q000378:metabolism*; D057225:Data Mining; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016680:Genome, Bacterial; D007700:Kinetics; D008353:Mannitol / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000090:Acetoacetates; D029968:Escherichia coli Proteins; D008353:Mannitol; C111982:ethyl 4-chloro-3-oxobutanoate; D009247:NADH, NADPH Oxidoreductases