Heme-containing enzymes represent one of nature's most versatile catalytic platforms, capable of mediating a broad spectrum of redox and group-transfer reactions. This review provides a comprehensive overview of the structural and mechanistic principles that underpin the bioengineering of heme proteins for both natural and abiological transformations. Here, we discuss that the core of reactivity of such metalloenzymes is the iron-porphyrin cofactor, whose tunable oxidation and spin states enable the formation of diverse high-energy intermediates, including iron-oxo, carbene, and nitrene species. Using recent bioengineering applications, we also show that the catalytic behavior of these intermediates is not solely determined by intrinsic cofactor chemistry but is profoundly influenced by the surrounding protein scaffold. Key factors such as axial ligation, second coordination sphere interactions, hydrogen-bonding networks, and local electric fields collectively govern substrate binding, intermediate stabilization, and reaction selectivity. The review further discusses a few applications of de novo protein design and artificial metalloenzymes that provide unprecedented control over active-site architecture, allowing the creation of highly robust and tunable catalysts.
Epari Sai Santosh Kumar, K. Dubey· FEBS Letters· 0 citations
Xenobiotic detoxification is the biotransformation of exogenous compounds entering the human body through enzymatic catalysis in order to assist their eventual elimination. In the past, static active-site models that mainly focus on substrate binding and oxidation chemistry have been employed to understand the xenobiotic functionality of CYP450s. However, modern computational approaches in the field of enzymology have changed this perspective, which show that the P450 machinery utilizes significant structural flexibility and electrostatic control to achieve its function. This review aims to unify these developments into an integrated mechanistic framework that encompasses substrate orientation, conformational gating, catalytic-site organization, transient water-channel assembly, and modulation of the electronic-structure landscape via classical electrostatic and steric confinement. Special focus has been given on how flexible loops/helices, gating residues, transient access channels, along with second-shell residues, dictate xenobiotic accommodation, regio-, stereo-, and chemoselectivity across various members of the CYP450 superfamily.
Vaibhav Bhatt, K. Dubey· ACS Bio & Med Chem Au· 0 citations
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