A General Tool for Engineering the NAD/NADP Cofactor Preference of Oxidoreductases.

ABSTRACT: The ability to control enzymatic nicotinamide cofactor utilization is critical for engineering efficient metabolic pathways. However, the complex interactions that determine cofactor-binding preference render this engineering particularly challenging. Physics-based models have been insufficiently accurate and blind directed evolution methods too inefficient to be widely adopted. Building on a comprehensive survey of previous studies and our own prior engineering successes, we present a structure-guided, semirational strategy for reversing enzymatic nicotinamide cofactor specificity. This heuristic-based approach leverages the diversity and sensitivity of catalytically productive cofactor binding geometries to limit the problem to an experimentally tractable scale. We demonstrate the effica

SUBMITTER: Cahn JK 

PROVIDER: S-EPMC8611728 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature