A Compound I Mimic Reveals the Transient Active Species of a Cytochrome P450 Enzyme: Insight into the Stereoselectivity of P450‐Catalysed Oxidations

Abstract

Catching the structure of cytochrome P450 enzymes in flagrante is crucial for the development of P450 biocatalysts, as most structures collected are found trapped in a precatalytic conformation. At the heart of P450 catalysis lies Cpd I, a short-lived, highly reactive intermediate, whose recalcitrant nature has thwarted most attempts at capturing catalytically relevant poses of P450s. We report the crystal structure of P450BM3 mimicking the state in the precise moment preceding epoxidation, which is in perfect agreement with the experimentally observed stereoselectivity. This structure was attained by incorporation of the stable Cpd I mimic oxomolybdenum mesoporphyrin IX into P450BM3 in the presence of styrene. The orientation of styrene to the Mo-oxo species in the crystal structures sheds light onto the dynamics involved in the rotation of styrene to present its vinyl group to Cpd I. This method serves as a powerful tool for predicting and modelling the stereoselectivity of P450 reactions.

Preparation of Dual‐activity Enzyme‐metal‐nanoparticles Conjugate Catalysts for Cascade Processes

“…In this sense, artificial metalloenzymes based on the insertion of organometallic complexes on the protein structure has been recently synthesized. [13][14][15][16][17][18][19] However, this required a timeconsuming in the preparation of the organometallic compound or required different modification of the protein for site-selective insertion of the metal. Another issue in this aspect is that mainly this artificial enzyme is water soluble as natural enzyme, presenting similar problems of compatibility in some experimental conditions or being required additional steps.…”

“…Therefore, alternative strategies, where both systems conserved full activity, are necessary. In this sense, artificial metalloenzymes based on the insertion of organometallic complexes on the protein structure has been recently synthesized [13–19] . However, this required a time‐consuming in the preparation of the organometallic compound or required different modification of the protein for site‐selective insertion of the metal.…”