Creating novel metabolic pathways by protein engineering for bioproduction.

A diverse array of natural products has been produced by cell biofactories through metabolic engineering, in which enzymes play essential roles in the complex metabolic network. However, the scope of such biotransformation can be limited by the capacities of natural enzymes. To broaden their scope, many natural enzymes have recently been engineered to activate non-native substrates and/or to employ new-to-nature reaction mechanisms, but most of these systems are only demonstrated for in vitro applications. To bridge the gap between in vitro and in vivo biocatalysis, we highlight recent progress in engineering enzymes with non-native substrates or new-to-nature mechanisms that have been successfully applied in living cells to create novel metabolic pathways.