title: [Cloning, expression and characterization of chiral alcohol dehydrogenase from Rhodococcus erythropolis ATCC 4277].
abstract: OBJECTIVE
  We characterized alcohol dehydrogenase from Rhodococcus erytropolis to catalyze ketoesters or ketones.
  
  
  METHODS
  We cloned alcohol dehydrogenase gene (adh) of 1047 bp from Rhodococcus erythropolis ATCC 4277, inserted the open reading frame of adh into vector pET-22b(+) and expressed in auto-inducing media for 24 h at 15 degrees C. The enzyme activity was determined at 30 degrees C using acetophenone as substrate.
  
  
  RESULTS
  Under the above conditions, the specific enzyme activity of crude extract was 2.6 U/mg. The optimal pH was between 6.0 and 6.5 and the enzyme can survived up to 60 degrees C. After incubation at 60 degrees C for 5 h, 80% enzyme activity remained. The optimal substrate among beta-ketoesters examined was ethyl acetoaetate. Ethyl 4-chloroacetoacetate was catalyzed by whole cell in aqueous phase. After chiral liquid chromatography, the product showed (R)-enantioselective.
  
  
  CONCLUSIONS
  The study shows that the enzyme might have potential in beta-ketoesters transformation on industrial scale.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing 211816, China. zqh021@yahoo.com.cn
authors.forename: Qinghe
authors.initials: Q
authors.lastname: Zhu
authors.forename: Honghua
authors.initials: H
authors.lastname: Jia
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Lisha
authors.initials: L
authors.lastname: Jia
authors.forename: Yingying
authors.initials: Y
authors.lastname: Ma
authors.forename: Ping
authors.initials: P
authors.lastname: Wei
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics*; D002712:Chlorides / Q000494:pharmacology; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D010957:Plasmids; D012240:Rhodococcus / Q000235:genetics*; D013237:Stereoisomerism; D013696:Temperature; D017967:Zinc Compounds / Q000494:pharmacology
chemical_list: D002712:Chlorides; D017967:Zinc Compounds; C016837:zinc chloride; D000426:Alcohol Dehydrogenase