Harnessing P450 Enzyme for Biotechnology and Synthetic Biology

Abstract

Cytochrome P450 enzymes (P450s, CYPs) catalyze the oxidative transformation of a wide range of organic substrates. Their functions are crucial to xenobiotic metabolism and steroid transformation in humans and other organisms. The enzymes are promising for synthetic biology applications but limited by several drawbacks including low turnover rates, poor stability, the dependance of expensive cofactors and redox partners, and the narrow substrate scope. To conquer these obstacles, emerging strategies including substrate engineering, usage of decoy and decoy‐based small molecules auxiliaries, designing of artificial enzyme cascades and the incorporation of materials have been explored based on the unique properties of P450s. These strategies can be applied to a wide range of P450s and can be combined with protein engineering to improve the enzymatic activities. This minireview will focus on some recent developments of these strategies which have been used to leverage P450 catalysis. Remaining challenges and future opportunities will also be discussed.

A Horseradish Peroxidase–Mediator System for Benzylic C–H Activation

“…Nonetheless, H 2 O 2 is an easy-to-use commodity chemical, and its use as a liquid reagent allows its concentration in the reaction solution to be controlled more easily than the concentration of O 2 . Our results complement recent studies using photocatalysts for converting alkylbenzenes to ketones, − as well as recent studies using cytochrome P450s, peroxygenase enzymes, and artificial metalloenzymes to convert alkylbenzenes to alcohols, ketones, or amines. − Relative to peroxygenase enzymes, the HRP–NHPI system offers the advantage of not requiring the substrate to fit within a confined active site, making it compatible for use with bulky substrates. Another practical advantage is the commercial availability of HRP, which avoids the need for recombinant enzyme expression and purification.…”

Enzyme Immobilization on a Delignified Bamboo Scaffold as a Green Hierarchical Bioreactor

“…Usually, lignin and hemicelluloses are degraded by H 2 O 2 in other wood-based supports; − no apparent deleterious effect was observed in our experiments, which implies that the delignified immobilized scaffolds have a predominant advantage when applied in this combined biosystem. Mutienzyme-based biotransformations have experienced rapid growth for both scientific and industrial applications in recent decades. − Conventionally, multienzymatic transformation is carried out in a one-pot reaction system. This approach has limitations due to potential cofactor competition, product mutual inhibition, etc .…”

A Unique P450 Peroxygenase System Facilitated by a Dual-Functional Small Molecule: Concept, Application, and Perspective

“…Despite being useful supplements to protein engineering, these strategies often play a role in combination with protein engineering. There have been many reviews discussing the abovementioned topics [34][35][36][37][38][39][40][41]. Herein, we focus on a unique H 2 O 2 -dependent P450 peroxygenase system facilitated by a dual functional small molecule (DFSM).…”