The Power of Deception: Using Decoy Molecules to Manipulate P450BM3 Biotransformations

Choose Your Own Adventure: A Comprehensive Database of Reactions Catalyzed by Cytochrome P450 BM3 Variants

“…Given the attractive features of P450 BM3, the enzyme has garnered significant attention and has been the subject of numerous reviews highlighting its exceptional properties and characteristics. Reviews have been conducted on various aspects of P450 BM3, including structural and mechanistic details, − hydroxylation, ,,− pharmaceutical compound hydroxylation, ,− epoxidation, ,,, and carbene/nitrene transfer. ,,,,,,− Additionally, studies have explored the use of decoy molecules to improve enzymatic activity, ,,,,,,− as well as P450 BM3-inorganic complexes. − Reviews of P450 BM3 frequently place it within a broader context of other P450 enzymes, resulting in a lack of focus. Though some reviews do describe reactions catalyzed by P450 BM3 variants, they are typically limited to discussion of the “best” variants for a given reaction and do not offer a comprehensive review of reported variants.…”

The Versatile Biocatalyst of Cytochrome P450 CYP102A1: Structure, Function, and Engineering

“…The aforementioned strategies are aimed at tuning the protein properties to fit substrates of interest. Conversely, chemical engineering optimizes substrate oxidation not by altering the enzyme but rather by fitting the substrate of interest into the active site of the WT CYP102A1 with a decoy molecule system [ 63 , 64 ] ( Figure 6 ). Taking advantage of the high oxidizing potential of Compound I, decoy molecules facilitate the generation of Compound I and the positioning of the substrate close to the heme iron for oxidation.…”

Hijacking Chemical Reactions of P450 Enzymes for Altered Chemical Reactions and Asymmetric Synthesis

“…As the name indicates, DFSM has two groups: an anchoring group, usually hydrophobic in nature, is connected to a basic group, which acts as a general acid-base catalyst to activate H 2 O 2 through an appropriate length of a linker (Figure 7B). Decoy and DFSM strategies have been extensively used to alter the substrate specificity and chemical reaction of both NADPH-and H 2 O 2 -dependent P450s, suggesting that these strategies, complemented by protein engineering technology and whole-cell catalysis, would help to generate a combination of toolboxes for developing practical P450 biocatalysts for the synthesis of novel and commercial chiral chemical products [7,8,[87][88][89][90][91][92][93][94][95][96][97][98][99][100][101][102][103] (Figure 7). Elsewhere, there are review articles that have extensively discussed decoy and DFSM strategies, as applied to different P450s to tune or switch the chemical reactions and to accept the non-natural substrates [6,7,96].…”

“…We also discuss how creating substrate-binding pockets and engineering subtle variations in the heme coordination residues convert hemoglobin into stereo-and regioselective biocatalysts. This review also reports recent advances in the manipulation of P450 chemical reactions using the substrate-mimicking decoy and dual-function small molecules for adapting the P450-mediated peroxide shunt pathway for regio-and stereoselective sulfoxidation, hydroxylation, epoxidation, and oxidative demethylation catalytic reactions for the costeffective use of biocatalysts [6,7]. Finally, we critically discuss the rational engineering of nicotinamide adenine dinucleotide phosphate (NADPH)-dependent monooxygenases into H 2 O 2 -peroxygenases, which uses low-cost H 2 O 2 compared to high-cost NADPH as the cofactor [8,9].…”