Engineering Cytochrome P450 Enzymes for Improved Activity towards Biomimetic 1,4‐NADH Cofactors

Compromising expensive taste: N-Benzyl-1,4-dihydronicotinamide is a NADH biomimic that is a cost-efficient alternative to NAD(P)H, but wild-type P450 BM-3 oxidation reactions are not supported by this NADH analogue. However, the activity of W1064S/R966D P450 BM-3 (see picture) with the biomimic is comparable to that with NADPH. The activity of P450cam with the analogue was also improved by site-directed mutagenesis. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.