title: Highly efficient synthesis of chiral alcohols with a novel NADH-dependent reductase from Streptomyces coelicolor.
abstract: An NADH-dependent reductase (ScCR) from Streptomyces coelicolor was discovered by genome mining for carbonyl reductases. ScCR was overexpressed in Escherichia coli BL21, purified to homogeneity and its catalytic properties were studied. This enzyme catalyzed the asymmetric reduction of a broad range of prochiral ketones including aryl ketones, α- and β-ketoesters, with high activity and excellent enantioselectivity (>99% ee) towards β-ketoesters. Among them, ethyl 4-chloro-3-oxobutanoate (COBE) was efficiently converted to ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE), an important pharmaceutical intermediate, in water/toluene biphasic system. As much as 600 g/L (3.6M) of COBE was asymmetrically reduced within 22 h using 2-propanol as a co-substrate for NADH regeneration, resulting in a yield of 93%, an enantioselectivity of >99% ee, and a total turnover number (TTN) of 12,100. These results indicate the potential of ScCR for the industrial production of valuable chiral alcohols.
authors.affiliation: Laboratory of Biocatalysis and Synthetic Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Wang
authors.forename: Chun-Xiu
authors.initials: CX
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Ni
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiang
authors.initials: X
authors.lastname: Liu
authors.forename: Jian-He
authors.initials: JH
authors.lastname: Xu
mesh_terms: D000090:Acetoacetates / Q000378:metabolism; D000438:Alcohols / Q000138:chemical synthesis* / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D010088:Oxidoreductases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012997:Solvents / Q000494:pharmacology; D013237:Stereoisomerism; D048372:Streptomyces coelicolor / Q000187:drug effects / Q000201:enzymology*; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature
chemical_list: D000090:Acetoacetates; D000438:Alcohols; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D012997:Solvents; D009243:NAD; C111982:ethyl 4-chloro-3-oxobutanoate; D004492:Edetic Acid; D010088:Oxidoreductases