Asymmetric reduction of alpha-ketoesters catalyzed by a 7-hydroxysteroid dehydyrogenase from Bacteroides fragilis

ORGN 91

Dunming Zhu, dzhu@smu.edu, Jenny Sterns, jsterns@smu.edu, Monica Ramirez, and Ling Hua, lhua@smu.edu. Department of Chemistry, Southern Methodist University, Dallas, TX 75275
Many efforts have been recently made to develop enzyme catalysts for the enantioselective reduction of ketones and varied level of success have been achieved. However, enzymatic reduction of bucky ketones such as aromatic alpha-ketoesters is much less successful than that of their small counterparts. Because hydroxysteroid dehydyrogenases takes bucky native substrates, we reasoned that this family of alcohol dehydrogenases might be promising in the reduction of bucky ketones. we successfully cloned a 7-hydroxysteroid dehydyrogenase gene from Bacteroides fragilis. The encoding protein was expressed in E. coli and purified from the cell-free extract. The purified enzyme was found to be active toward a series of aromatic alpha-ketoesters. This alcohol dehydrogenase was also active toward alpha-ketoesters with bucky groups such as tert-butyl and cyclohexyl groups at the other side of carbonyl group. The effects of substituents at the benzyl ring of aromatic alpha-ketoesters on the activity and enantioselectivity were studied. This enzyme was then applied to the synthesis of a few aromatic a-hydroxy carboxylic acid esters of pharmaceutical importance.