Biochemical characterization and substrate profile of a highly enantioselective carbonyl reductase from Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 169-179, 2013.
Article
in Chinese
| WPRIM
| ID: wpr-233257
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pichia
/
Stereoisomerism
/
Substrate Specificity
/
Temperature
/
Biotechnology
/
Recombinant Proteins
/
Molecular Sequence Data
/
Chemistry
/
Amino Acid Sequence
/
Cloning, Molecular
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2013
Type:
Article
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