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[Cloning and characterization of a novel carbonyl reductase for asymmetric reduction of bulky diaryl ketones].
|Title||[Cloning and characterization of a novel carbonyl reductase for asymmetric reduction of bulky diaryl ketones].|
|Publication Type||Journal Article|
|Year of Publication||2013|
|Authors||Li Z, Liu W, Chen X, Jia S, Wu Q, Zhu D, Ma Y|
|Journal||Sheng wu gong cheng xue bao = Chinese journal of biotechnology|
|Date Published||2013 Jan|
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.
|Alternate Journal||Sheng Wu Gong Cheng Xue Bao|