Cloning and characterization of a novel carbonyl reductase for asymmetric reduction of bulky diaryl ketones.
- Author:
Zhe LI
1
;
Weidong LIU
;
Xi CHEN
;
Shiru JIA
;
Qiaqing WU
;
Dunming ZHU
;
Yanhe MA
Author Information
1. College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, China.
- Publication Type:Journal Article
- MeSH:
Alcohol Oxidoreductases;
genetics;
Amino Acid Sequence;
Catalysis;
Cloning, Molecular;
Ketones;
chemistry;
Molecular Sequence Data;
Pichia;
enzymology;
genetics;
Stereoisomerism
- From:
Chinese Journal of Biotechnology
2013;29(1):68-77
- CountryChina
- Language:Chinese
-
Abstract:
Asymmetric reduction of bulky diaryl ketones is still one of the challenging tasks in biocatalysis. By genomic data mining, a putative carbonyl reductase gene pascr was found in Pichia pastoris GS115. pascr was cloned and over-expressed in Escherichia coli Rosseta2 (DE3). The recombinant enzyme was purified to homogeneity by Ni-NTA column and its catalytic properties were studied. PasCR strictly used NADPH as cofactor, gel filtration and SDS-PAGE analysis suggested that the native form of PasCR was a dimmer. PasCR exhibited the highest activity at 35 degrees C in phosphate buffer at pH 6.5. The enzyme catalyzed the reduction of some bulky diaryl ketones, such as 4-methylbenzophenone, 2-methylbenzophenone and 4-chlorobenzophenone, especially for 4-methylbenzophenone, the product S--alcohol was obtained with 85% ee.
