title: [Cloning and characterization of a novel carbonyl reductase for asymmetric reduction of bulky diaryl ketones].
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.
authors: College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, China.
authors: Zhe
authors: Z
authors: Li
authors: Weidong
authors: W
authors: Liu
authors: Xi
authors: X
authors: Chen
authors: Shiru
authors: S
authors: Jia
authors: Qiaqing
authors: Q
authors: Wu
authors: Dunming
authors: D
authors: Zhu
authors: Yanhe
authors: Y
authors: Ma
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics*; D000595:Amino Acid Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D007659:Ketones / Q000737:chemistry*; D008969:Molecular Sequence Data; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D013237:Stereoisomerism
chemical_list: D007659:Ketones; D000429:Alcohol Oxidoreductases