title: Stereoselective reduction of ethyl 4-chloro-3-oxobutanoate by Escherichia coli transformant cells coexpressing the aldehyde reductase and glucose dehydrogenase genes.
abstract: The asymmetric reduction of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (R)-4-chloro-3-hydroxybutanoate [(R)-CHBE] using Escherichia coli cells, which coexpress both the aldehyde reductase gene from Sporobolomyces salmonicolor and the glucose dehydrogenase (GDH) gene from Bacillus megaterium as a catalyst was investigated. In an organic solvent-water two-phase system, (R)-CHBE formed in the organic phase amounted to 1610 mM (268 mg/ml), with a molar yield of 94.1% and an optical purity of 91.7% enantiomeric excess. The calculated turnover number of NADP+ to CHBE formed was 13,500 mol/mol. Since the use of E. coli JM109 cells harboring pKAR and pACGD as a catalyst is simple, and does not require the addition of GDH or the isolation of the enzymes, it is highly advantageous for the practical synthesis of (R)-CHBE.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kataoka
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Kawabata
authors.forename: M
authors.initials: M
authors.lastname: Wada
authors.forename: K
authors.initials: K
authors.lastname: Kita
authors.forename: H
authors.initials: H
authors.lastname: Yanase
authors.forename: S
authors.initials: S
authors.lastname: Shimizu
mesh_terms: D000090:Acetoacetates / Q000378:metabolism*; D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism*; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D003904:Mitosporic Fungi / Q000201:enzymology / Q000235:genetics; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D013237:Stereoisomerism; D014169:Transformation, Bacterial
chemical_list: D000090:Acetoacetates; C111982:ethyl 4-chloro-3-oxobutanoate; D005948:Glucose Dehydrogenases; D000449:Aldehyde Reductase; D042843:Glucose 1-Dehydrogenase