An Overview of Cytochrome P450 Immobilization Strategies for Drug Metabolism Studies, Biosensing, and Biocatalytic Applications: Challenges and Opportunities

Cytochrome P450s (P450s or CYPs) are a large superfamily of ubiquitous heme proteins. Consistent with their natural roles in oxidative metabolism of a broad range of substrates including antibiotics and xenobiotics and in the biosynthesis of complex natural products, there are two main biotechnological applications: drug metabolism studies and biocatalytic transformations. Metabolic analyses are typically performed with the P450 enzyme immobilized on a biosensing platform. Biocatalytic reactions also increasingly make use of immobilization methods with the goal of improving operational stability of the P450s. In this Review, we briefly introduce the growing field of enzyme immobilization then focus on the advances, benefits, and challenges that apply specifically to P450s. Classical immobilization methods are presented, and recently developed materials and strategies that address the need for operational stability, the multicomponent structure of P450s, their requirement for a source of electrons, and the need to overcome limited O2 availability are discussed in the context of metabolic, biosensing, and biocatalytic applications.