8WS4

Crystal structure of the CYP199A4 mutant F182A in complex with 4-methoxybenzoic acid

Summary for 8WS4

• Entry DOI: 10.2210/pdb8ws4/pdb
• Descriptor: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE, 4-METHOXYBENZOIC ACID, ... (5 entities in total)
• Functional Keywords: cyp199a4, complex, oxidoreductase
• Biological source: Rhodopseudomonas palustris HaA2
• Total number of polymer chains: 1
• Total formula weight: 46598.40

Authors

Jiang, Y.,Cong, Z. (deposition date: 2023-10-16, release date: 2024-10-16)

Primary citation

Zhao, P.,Jiang, Y.,Wang, Q.,Chen, J.,Yao, F.,Cong, Z.
Crucial gating residues govern the enhancement of peroxygenase activity in an engineered cytochrome P450 O -demethylase.
Chem Sci, 15:8062-8070, 2024

PubMed Abstract: P450-catalyzed -demethylation reactions have recently attracted particular attention because of their potential applications in lignin bioconversion. We recently enabled the peroxygenase activity of CYP199A4, a NADH-dependent cytochrome P450 monooxygenase from , by engineering a hydrogen peroxide (HO) tunnel. In this report, we reveal by crystallography and molecule dynamics simulations that key residues located at one of the water tunnels in CYP199A4 play a crucial gating role, which enhances the peroxygenase activity by regulating the inflow of HO. These results provide a more complete understanding of the mechanism by which monooxygenase is converted into peroxygenase activity through the HO tunnel engineering (HTE) strategy. Furthermore, a library of engineered CYP199A4 peroxygenases was constructed to explore their application potentials for -demethylation of various methoxy-substituted benzoic acid derivatives. The engineered CYP199A4 peroxygenases showed good functional group tolerance and preferential -demethylation at the - or -position, indicating potential -demethylation of H- and G-type lignin monomers. This work reveals the feasibility of the HTE strategy in creating P450 peroxygenase from a mechanistic perspective, laying the foundation for developing an effective P450 -demethylase applicable in lignin bioconversion.

PubMed: 38817576
DOI: 10.1039/d4sc02418d

Experimental method

X-RAY DIFFRACTION (1.53 Å)