Surface-Binding Peptide Facilitates Electricity-Driven NADPH-Free Cytochrome P450 Catalysis

Industrial biotechnology aims to exploit cytochrome P450 enzymes to access their sophisticated catalytic activity for challenging chemical reactions on inert C-H bonds. Limited by the need for NADPH, approaches to bind P450 enzymes to electrode surfaces for an artificial electron supply are promising. Here, we demonstrate that a recombinant fusion of an indium tin oxide binding peptide and the multi-domain class VIII cytochrome P450 BM3 can be used in electrically driven catalysis. Bioelectrocatalytic activity is analyzed by direct product quantification resulting in superior activity of the specifically immobilized P450 BM3 in contrast to unspecifically adsorbed enzyme. Spacer and anchor point studies imply that enzyme flexibility and alignment are crucial factors to achieve high activity on the electrode. Furthermore, we demonstrate that our approach is also feasible for pharmaceutical application using naringenin as substrate.