Immobilization of a Bacterial Cytochrome P450 Monooxygenase System on a Solid Support

Bacterial cytochrome P450s (P450s), which catalyze regio‐ and stereoselective oxidations of hydrocarbons with high turnover rates, are attractive biocatalysts for fine chemical production. Enzyme immobilization is needed for cost‐effective industrial manufacturing. However, immobilization of P450s is difficult because electron‐transfer proteins are involved in catalysis and anchoring these can prevent them from functioning as shuttle molecules for carrying electrons. We studied a heterotrimeric protein‐mediated co‐immobilization of a bacterial P450, and its electron‐transfer protein and reductase. Fusion with subunits of a heterotrimeric Sulfolobus solfataricus proliferating cell nuclear antigen (PCNA) enabled immobilization of the three proteins on a solid support. The co‐immobilized enzymes catalyzed monooxygenation because the electron‐transfer protein fused to PCNA via a single peptide linker retained its electron‐transport function. Immobilisierung mehrerer Enzyme: Bakterielle Cytochrom‐P450‐Enzyme sind auf Elektronen angewiesen, die über Elektronentransferproteine von NAD(P)H‐abhängigen Reduktasen zur Verfügung gestellt werden. Die proteinvermittelte Bildung eines Heterotrimers aus P450, Elektronentransferprotein und Reduktase auf einem festen Träger bringt die drei Proteine in direkte Nähe zueinander und ermöglicht den Elektronentransfer zwischen P450 und der Reduktase.