A High-Throughput Screening Method for Chiral Alcohols and its Application to Determine Enantioselectivity of Lipases and Esterases

A new colorimetric method to determine the enantiopurity of chiral alcohols is presented based on enantioselective oxidation by two alcohol dehydrogenases. The oxidation of either (R)- or (S)-alcohol produces NADPH (or NADH), which reduces 2-(4-iodophenyl)-3-(4- nitrophenyl)-5-phenyl-2H-tetrazolium to its corresponding formazan derivative. Application of this method to screen lipase variant libraries for enantioselective hydrolysis is also described.