title: A novel NADH-dependent carbonyl reductase from Kluyveromyces aestuarii and comparison of NADH-regeneration system for the synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate.
abstract: To compare NADH-regeneration systems for the synthesis of (S)-4-chloro-3-hydroxybutanoate (ECHB), a novel NADH-dependent carbonyl reductase (KaCR1), which reduced ethyl 4-chloroacetoacetate (ECAA) to form (S)-ECHB, was screened and purified from Kluyveromyces aestuarii and a gene encoding KaCR1 was cloned. Glucose dehydrogenase (GDH) and formate dehydrogenase (FDH) were compared as enzymes for NADH regeneration using Escherichia coli cells coexpressing each enzyme with KaCR1. E. coli cells coexpressing GDH produced 45.6 g/l of (S)-ECHB from 50 g/l of ECAA and E. coli cells coexpressing FDH, alternatively, produced only 19.0 g/l. The low productivity in the case of FDH was suggested to result from the low activity and instability of FDH.
authors.affiliation: Life Science Development Center, CPI Company, Daicel Chemical Industries, Ltd., Tsukuba Research Center, Ibaraki, Japan. h-yamamoto@daicel.co.jp
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Kazuya
authors.initials: K
authors.lastname: Mitsuhashi
authors.forename: Norihiro
authors.initials: N
authors.lastname: Kimoto
authors.forename: Akinobu
authors.initials: A
authors.lastname: Matsuyama
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Esaki
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kobayashi
mesh_terms: D000090:Acetoacetates / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002087:Butyrates / Q000378:metabolism*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism; D005560:Formate Dehydrogenases / Q000378:metabolism; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism; D007716:Kluyveromyces / Q000201:enzymology*; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
chemical_list: D000090:Acetoacetates; D002087:Butyrates; C121244:ethyl 4-chloro-3-hydroxybutanoate; D009243:NAD; C111982:ethyl 4-chloro-3-oxobutanoate; D000429:Alcohol Oxidoreductases; D042843:Glucose 1-Dehydrogenase; D005560:Formate Dehydrogenases; C020644:Formate dehydrogenase (cytochrome); C036543:short chain trans-2-enoyl-CoA reductase; D009247:NADH, NADPH Oxidoreductases; D064429:Fatty Acid Synthases