Asymmetric Synthesis Using Cofactor-Requiring Enzymes

The use of cofactor-requiring enzymes as catalysts for large-scale requires efficient and economical procedures for in situ regeneration of these cofactors. This manuscript summarizes the procedures which are now available for cofactor preparation and regeneration. ATP can be effectively regenerated from ADP (and AMP) using acetyl phosphate and acetate kinase (and adenylate kinase), and it can be prepared inexpensively from RNA. Use of ATP-requiring enzymes is now routine (at least as far as the ATP regeneration is concerned). The use of the nicotinamide cofactors is more difficult, because these materials decompose in solution. The best procedure for regenerating NAD(P)H from NAD(P)+ are those based on formate/formate dehydrogenase, glucose 6-phosphate/glucose-6-phosphate dehydrogenase, and ethanol/alcohol dehydrogenase/aldehyde dehydrogenase. The best procedures for regenerating NAD(P)+ from NAD(P)H use dioxygen/methyl viologen or ketoglutarate/glutamic dehydrogenase.