Reetz_2000_Pure.Appl.Chem_72_1615

A novel approach to developing enantioselective enzymes for use in organic chem-istry has been devised which is independent of structural or mechanistic aspects. The under-lying idea is to combine appropriate methods of random mutagenesis, gene expression, andhigh-throughput screening for enantioselectivity. If these actions are performed in repetitivecycles, an evolutionary pressure is created that leads to sequential improvements of the enan-tioselectivity of a given enzyme-catalyzed reaction. The concept is illustrated by an exampleinvolving the lipase-catalyzed hydrolytic kinetic resolution of an alpha-chiral ester, the enantio-selectivity increasing from ee= 2% (E= 1.1) for a wild-type enzyme to ee= 90-93% (E=25) for the best mutants