Highly Enantioselective Mutant Carbonyl Reductases Created via Structure Based Site Saturation Mutagenesis英文文献.pdfVIP

pubs.acs.org/joc Highly Enantioselective Mutant Carbonyl Reductases Created via Structure-Based Site-Saturation Mutagenesis Hongmei Li,‡ Yan Yang,‡ Dunming Zhu,*,†,‡ Ling Hua,‡,§ and Katherine Kantardjieff^ †State Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China, ‡Department of Chemistry, Southern Methodist University, Dallas, Texas 75275, United States, § China Research Center, Genencor, A Danisco Division, Shanghai 200335, China, and ^Department of Chemistry, California State Polytechnic University, Pomona, California 91768, United States zhu_dm@tib.cas.cn Received August 6, 2010 A carbonyl reductase from Sporobolomyces salmonicolor reduced para-substituted acetophenones with low enantioselectivity. Enzyme-substrate docking studies revealed that residues M242 and Q245 were in close proximity to the para-substituent of acetophenones in the substrate binding site. Site-saturation mutagen- esis of M242 or Q245, and double mutation of M242 and Q245 were performed in order to enhance the enzyme’s enantioselectivity toward the reduction of para-substituted acetophenones. Three Q245 mutants were obtained, which inverted the enantiopreference of product alcohols from (R)- to (S)-configuration with high ee values (Org. Lett. 2008, 10, 525-528). Four M242 mutant enzymes also showed greater preference for the formation