Cytochromes P450 in biosensing and biosynthesis applications: Recent progress and future perspectives

Cytochrome P450 (CYP450s) as a large superfamily of ubiquitous heme-containing monooxygenases are widely distributed in kingdoms of life, catalyzing versatile reactions that introduce oxygen into a vast range of molecules. To improve the activity, stability and catalytic efficiency, the methods for immobilizing CYP450s on a biosensing platform and biosynthesis of complex natural products were reported. However, these biotechnologies are hampered by the limited O 2 availability. In this review, we summarize and highlight a panorama of the classical methods for biosensor, nanoreactor with confinement function and cascade reactions, bioimaging for detecting CYP450s activity and biosynthesis of complex natural and unnatural products. Then, the gas-solid-liquid interface (triphase interface) is introduced for CYP450s immobilization to overcome limited O 2 availability and drive the catalytic cycle of CYP450s-H 2 O 2 system by in situ generating H 2 O 2 and controlling interfacial wettability, which affords a convenient way for their synthetic application. • We give an overview of the strategies of CYP450s catalysis and reactor fabrication for applications extending from biocatalysis, bioimaging to other new emerging fields. • The fabrication of CYP450s catalytic reactors with controlled diffusion are presented. • The triphase interface for CYP450s immobilization to address the limited O 2 availability. • The catalytic efficiency of the peroxide-shunt pathway of CYP450s (hydrogen peroxide as co-substrate) could also be largely boosted by sufficiently O 2 supply and generated H 2 O 2 with the novel triphase interface reactor.