title: Biocatalytic characterization of a short-chain alcohol dehydrogenase with broad substrate specificity from thermophilic Carboxydothermus hydrogenoformans.
abstract: The gene encoding a novel short-chain alcohol dehydrogenase in the thermophilic bacterium, Carboxydothermus hydrogenoformans, was identified and overexpressed in Escherichia coli. The enzyme was thermally stable and displayed the highest activity at 70 °C and pH 6.0. It preferred NAD(H) over NADP(H) as a cofactor and exhibited broad substrate specificity towards aliphatic ketones, cycloalkanones, aromatic ketones, and ketoesters. Furthermore, ethyl benzoylformate was asymmetrically reduced by the purified enzyme, using an additional coupled NADH regeneration system, with 95 % conversion and in an enantiomeric excess of (99.9 %). The results of this study may lead to the discovery of a novel method for asymmetric reduction of alcohols, which is an important tool in organic synthesis.
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University,  402-Building D, 1378 West Wenyi Road, Hangzhou, 311121, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhou
authors.forename: Shun-Cheng
authors.initials: SC
authors.lastname: Zhang
authors.forename: Dun-Yue
authors.initials: DY
authors.lastname: Lai
authors.forename: Shuang-Ling
authors.initials: SL
authors.lastname: Zhang
authors.forename: Zhen-Ming
authors.initials: ZM
authors.lastname: Chen
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D003001:Cloning, Molecular; D003067:Coenzymes / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006094:Gram-Positive Bacteria / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009243:NAD / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D003067:Coenzymes; D011994:Recombinant Proteins; D009243:NAD; D000426:Alcohol Dehydrogenase