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[Biochemical characterization and substrate profile of a highly enantioselective carbonyl reductase from Pichia pastoris].
- Source :
-
Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2013 Feb; Vol. 29 (2), pp. 169-79. - Publication Year :
- 2013
-
Abstract
- Carbonyl reductases catalyze carbonyl compounds to chiral alcohols that are important building blocks in fine chemical industry. To study carbonyl reductase from Pichia pastoris GS115 (ppcr), we discovered a new gene (ppcr) encoding an NADPH-dependent carbonyl reductase by genomic data mining. It was amplified by PCR from the genomic DNA, and expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to homogeneity. The optimum temperature was 37 degrees C and the optimum pH of PPCR was 6.0. PPCR was stable below 45 degrees C. The Km and k(cat) value of the enzyme for ethyl 3-methyl-2-oxobutanoate were 9.48 mmol/L and 0.12 s, respectively. The enzyme had broad substrate specificity and high enantioselectivity. It catalyzed the reduction of aldehydes, a-ketoesters, beta-ketoesters and aryl ketones to give the corresponding alcohols with >97% ee with only a few exceptions, showing its application potential in the synthesis of chiral alcohols.
- Subjects :
- Alcohol Oxidoreductases chemistry
Alcohol Oxidoreductases genetics
Amino Acid Sequence
Biotechnology methods
Cloning, Molecular
Escherichia coli genetics
Escherichia coli metabolism
Molecular Sequence Data
Recombinant Proteins chemistry
Recombinant Proteins genetics
Stereoisomerism
Substrate Specificity
Temperature
Alcohol Oxidoreductases biosynthesis
Pichia enzymology
Recombinant Proteins biosynthesis
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-3061
- Volume :
- 29
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 23697162