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

Abstract

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 O 2 availability are discussed in the context of metabolic, biosensing, and biocatalytic applications.

Curcuminoids for Metabolic Syndrome: Meta-Analysis Evidences Toward Personalized Prevention and Treatment Management

“…Drug metabolism is categorized into phase I and phase II metabolism. Cytochrome P450 (CYP) enzymes, a large superfamily of heme proteins, play a major role in modulating phase I metabolism by catalyzing oxidative biotransformation of drugs and xenobiotics ( 165 , 166 ). At least 57 CYP genes have been identified in humans.…”

“…At least 57 CYP genes have been identified in humans. Of 18 families and 44 subfamilies human CYPs, only families 1–3 are most active in hepatic metabolism with five of them (CYP 1A2, 2C9, 2C19, 2D6, and 3A4) contributing to metabolism of 75% of drugs ( 165 , 167 ). Drug therapy can induce or inhibit CYP enzymes resulting in drug-drug interaction.…”

Co-Crystal Structure-Guided Optimization of Dual-Functional Small Molecules for Improving the Peroxygenase Activity of Cytochrome P450BM3

“…Cytochrome P450 enzymes (P450s) are a superfamily of heme proteins involved in drug metabolism, xenobiotic detoxification, and steroid biosynthesis [ 1 , 2 ]. P450s are promising biocatalysts for organic synthesis, drug discovery, and bioremediation because of their versatile catalytic oxyfunctionalizations of a variety of substrates [ 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ].…”

Immobilization-Stabilization of β-Glucosidase for Implementation of Intensified Hydrolysis of Cellobiose in Continuous Flow Reactors

“…Enzyme immobilization is an enabling technology that facilitates enzyme handling, permits its cyclic reuse and continuous use, and, if properly designed, might contribute to increasing the operational stability of the enzyme [19,20]. Immobilization design involves the choice of material support (where the enzyme is immobilized), the immobilization chemistry and the conditions of immobilization [19,[21][22][23]. As a result, different properties can be modulated suited for applications, e.g., reactor format suitability, activity, stability, and practical use.…”

“…Studies mainly involve the immobilization into preexisting materials and the evaluation of the influence of the material supports in the interplay with different immobilization chemistries. Catalytic properties as recovered activity and stability were evaluated [18][19][20][21][22][23][24][25][26]. Active immobilization by adsorption (anion exchange based ionic adsorption) or covalent binding (via aldehyde chemistry) to porous material supports have been shown as a promising strategy to obtain active and stable catalysts of different βglucosidases.…”