A novel zinc-containing alpha-keto ester reductase from actinomycete: an approach based on protein chemistry and bioinformatics.
We have achieved the purification of an alpha-keto ester reductase (SCKER) from S. coelicolor A3(2) whole cells. SCKER proved to be a homotetramer of 132 kDa containing one equivalent of zinc ion per subunit. The enzyme differed from other alpha-keto ester reductases from microorganisms with regard to subunit structure and metal ion dependency. From a computer search using the protein data banks, the N-terminal amino acid sequence of SCKER was consistent with that of a possible zinc containing alcohol dehydrogenase in S. coelicolor A3(2). None of three hypothetical proteins of S. coelocor A3(2) having a high homology sequence with those of already purified alpha-keto ester reductases from S. thermocyaneoviolaceus [Yamaguchi, H., et al., Biosci. Biotechnol. Biochem., 66, 588-597 (2002)] was identical with that of SCKER.
Ishihara
Kohji
K
Department of Life Science, Okayama University of Science, Okayama 700-0005, Japan. ishihara@as.ous.ac.jp
Yamaguchi
Hitomi
H
Omori
Taketo
T
Uemura
Tadashi
T
Nakajima
Nobuyoshi
N
Esaki
Nobuyoshi
N
D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D003001:Cloning, Molecular; D019295:Computational Biology; D048372:Streptomyces coelicolor / Q000201:enzymology*; D013379:Substrate Specificity / Q000502:physiology; D015032:Zinc / Q000378:metabolism*
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; D015032:Zinc