(719d) Probing the Synthetic Nicotinamide Cofactor-Enzyme Interface for Redox Biocatalysis

Redox biotransformations have been utilized in the production of several valuable chemicals with the increased availability of characterized oxidoreductases. However, these processes require the native redox cofactors-- NAD(P)H-- which act as a bottle neck due to their high costs and poor stability. To circumvent this problem, much attention has been drawn to fully synthetic nicotinamide analogues due to their ease of synthesis and increased stability. However, due to their large deviance in structure from their native counterparts, there have only been limited attempts to successfully engineer oxidoreductases for the use of these synthetic nicotinamide cofactors.

We present an approach where we systematically engineer the synthetic nicotinamide cofactor-enzyme interface through the simultaneous modification of different synthetic cofactor functional groups, such as length/flexibility, polar contact and regiospecificity of said polar contact, alongside rational design of new interactions on the enzyme active site. Here, we demonstrate that in order to generate the ideal cofactor-enzyme pair to drive redox catalysis with synthetic nicotinamide cofactors, it will be necessary to use an approach from both the enzyme and cofactor front.