title: Cloning and overexpression of an NADH-dependent alcohol dehydrogenase gene from Candida maris involved in (R)-selective reduction of 5-acetylfuro[2,3-c]pyridine.
abstract: 5-((R)-1-Hydroxyethyl)-furo[2,3-c]pyridine ((R)-FPH) is a useful chiral building block in the synthesis of pharmaceuticals. An NADH-dependent alcohol dehydrogenase (AFPDH) isolated from Candida maris catalyzed the reduction of 5-acetylfuro[2,3-c]pyridine (AFP) to (R)-FPH with 100% enantiomeric excess. The gene encoding AFPDH was cloned and sequenced. The AFPDH gene comprises 762 bp and encodes a polypeptide of 27,230 Da. The deduced amino acid sequence showed a high degree of similarity to those of other members of the short-chain alcohol dehydrogenase superfamily. The AFPDH gene was overexpressed in Escherichia coli under the control of the lac promoter. One L of the cultured broth of an E. coli transformant coexpressing AFPDH and the glucose dehydrogenase (GDH) gene reduced 250 g of AFP to (R)-FPH in an organic solvent two-phase system. Under coupling with NADH regeneration using 2-propanol, 1 L of the cultured broth of an E. coli transformant expressing the AFPDH gene reduced 150 g of AFP to (R)-FPH. The optical purity of the (R)-FPH formed was 100% enantiomeric excess under both reaction conditions.
authors: Frontier Biochemical and Medical Research Laboratories, Kaneka Corporation, Takasago, Hyogo, Japan. Shigeru.Kawano@kaneka.co.jp
authors: Shigeru
authors: S
authors: Kawano
authors: Miho
authors: M
authors: Yano
authors: Junzo
authors: J
authors: Hasegawa
authors: Yoshihiko
authors: Y
authors: Yasohara
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011725:Pyridines / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: C570734:5-((R)-1-hydroxyethyl)-furo(2,3-c)pyridine; C570733:5-acetylfuro(2,3-c)pyridine; D011725:Pyridines; D000429:Alcohol Oxidoreductases; C084155:alcohol dehydrogenase (NAD(P)+)