6L69

Crystal structure of CYP154C2 from Streptomyces avermitilis

Summary for 6L69

• Entry DOI: 10.2210/pdb6l69/pdb
• Descriptor: Cytochrome P450 hydroxylase, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• Functional Keywords: cytochrome p450, substrate-free, hydroxylase, oxidoreductase
• Biological source: Streptomyces avermitilis (strain ATCC 31267 / DSM 46492 / JCM 5070 / NBRC 14893 / NCIMB 12804 / NRRL 8165 / MA-4680)
• Total number of polymer chains: 2
• Total formula weight: 91777.80

Authors

Xu, L.H.,Fushinobu, S. (deposition date: 2019-10-28, release date: 2020-09-23, Last modification date: 2023-11-22)

Primary citation

Wang, Q.,Ma, B.,Fushinobu, S.,Zhang, C.,Xu, L.H.
Regio- and stereoselective hydroxylation of testosterone by a novel cytochrome P450 154C2 from Streptomyces avermitilis.
Biochem.Biophys.Res.Commun., 522:355-361, 2020

PubMed Abstract: Cytochrome P450 enzymes (P450 or CYP) are some of the most versatile biocatalysts, and offer advantages for oxidizing unreactive C-H bonds in mild conditions. In this study, we identified a novel cytochrome P450 154C2 from Streptomyces avermitilis and characterized its function in 2α-hydroxylation of testosterone with regio- and stereoselectivity. To investigate the efficiency of electron transfer, we conducted biotransformation using two different P450 redox partners-RhFRED (RhF reductase domain) from Rhodococcus sp. and Pdx (putidaredoxin)/Pdr (putidaredoxin reductase) from Pseudomonas putida and revealed that RhFRED was more effective than Pdx/Pdr, especially in vivo. The K and k values for testosterone were estimated to be 0.16 ± 0.05 mM and 0.13 ± 0.02 min, and k/K was 0.81 min mM. We also determined the crystal structure of the substrate-free form of CYP154C2 at 1.5 Å resolution. The structure has a closed conformation, and the substrate binding pocket is narrow, which can explain the strict substrate specificity of the enzyme.

PubMed: 31767148
DOI: 10.1016/j.bbrc.2019.11.091

Experimental method

X-RAY DIFFRACTION (1.5 Å)