Download MendeleyEnabling Peroxygenase Activity in Cytochrome P450 Monooxygenases by Engineering Hydrogen Peroxide Tunnels.
Zhao, P., Kong, F., Jiang, Y., Qin, X., Tian, X., Cong, Z.(2023) J Am Chem Soc 145: 5506-5511
- PubMed: 36790023
- DOI: https://doi.org/10.1021/jacs.3c00195
- Primary Citation of Related Structures:
8HGC, 8HGT - PubMed Abstract:
Given prominent physicochemical similarities between H 2 O 2 and water, we report a new strategy for promoting the peroxygenase activity of P450 enzymes by engineering their water tunnels to facilitate H 2 O 2 access to the heme center buried therein. Specifically, the H 2 O 2 -driven activities of two native NADH-dependent P450 enzymes (CYP199A4 and CYP153A M.aq ) increase significantly (by >183-fold and >15-fold, respectively). Additionally, the amount of H 2 O 2 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 H 2 O 2 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 H 2 O 2 tunnel engineering, thus broadening the application scope of P450s in synthetic chemistry and synthetic biology.
Organizational Affiliation:
CAS Key Laboratory of Biofuels and Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, China.
