8HGT

Crystal structure of the CYP153A mutant V456A from Marinobacter aquaeolei

Summary for 8HGT

• Entry DOI: 10.2210/pdb8hgt/pdb
• Descriptor: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• Functional Keywords: cyp153a, oxidoreductase
• Biological source: Marinobacter nauticus
• Total number of polymer chains: 1
• Total formula weight: 55867.47

Authors

Jiang, Y.,Tian, X.,Cong, Z. (deposition date: 2022-11-15, release date: 2023-08-16)

Primary citation

Zhao, P.,Kong, F.,Jiang, Y.,Qin, X.,Tian, X.,Cong, Z.
Enabling Peroxygenase Activity in Cytochrome P450 Monooxygenases by Engineering Hydrogen Peroxide Tunnels.
J.Am.Chem.Soc., 145:5506-5511, 2023

PubMed Abstract: Given prominent physicochemical similarities between HO and water, we report a new strategy for promoting the peroxygenase activity of P450 enzymes by engineering their water tunnels to facilitate HO access to the heme center buried therein. Specifically, the HO-driven activities of two native NADH-dependent P450 enzymes (CYP199A4 and CYP153A) increase significantly (by >183-fold and >15-fold, respectively). Additionally, the amount of HO required for an artificial P450 peroxygenase facilitated by a dual-functional small molecule to obtain the desired product is reduced by 95%-97.5% (with ∼95% coupling efficiency). Structural analysis suggests that mutating the residue at the bottleneck of the water tunnel may open a second pathway for HO to flow to the heme center (in addition to the natural substrate tunnel). This study highlights a promising, generalizable strategy whereby P450 monooxygenases can be modified to adopt peroxygenase activity through HO tunnel engineering, thus broadening the application scope of P450s in synthetic chemistry and synthetic biology.

PubMed: 36790023
DOI: 10.1021/jacs.3c00195

Experimental method

X-RAY DIFFRACTION (2.06 Å)