6A7I

CYP154C4 from Streptomyces sp. W2061

Summary for 6A7I

• Entry DOI: 10.2210/pdb6a7i/pdb
• Descriptor: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• Functional Keywords: cytochrome p450, streptomyces, steroid hydroxylase, oxidoreductase
• Biological source: Streptomyces sp. JS01
• Total number of polymer chains: 1
• Total formula weight: 45392.36

Authors

Lee, C.W.,Lee, J.H. (deposition date: 2018-07-03, release date: 2019-01-16, Last modification date: 2024-03-27)

Primary citation

Dangi, B.,Lee, C.W.,Kim, K.H.,Park, S.H.,Yu, E.J.,Jeong, C.S.,Park, H.,Lee, J.H.,Oh, T.J.
Characterization of two steroid hydroxylases from different Streptomyces spp. and their ligand-bound and -unbound crystal structures.
Febs J., 286:1683-1699, 2019

PubMed Abstract: Bacterial cytochrome P450 (CYP) enzymes are involved in the hydroxylation of various endogenous substrates while using a heme molecule as a cofactor. CYPs have gained biotechnological interest as useful biocatalysts capable of altering chemical structures by adding a hydroxyl group in a regiospecific manner. Here, we identified, purified, and characterized two CYP154C4 proteins from Streptomyces sp. W2061 (StCYP154C4-1) and Streptomyces sp. ATCC 11861 (StCYP154C4-2). Activity assays showed that both StCYP154C4-1 and StCYP154C4-2 can produce 2'-hydroxylated testosterone, which differs from the activity of a previously described NfCYP154C5 from Nocardia farcinica in terms of its 16α-hydroxylation of testosterone. To better understand the molecular basis of the regioselectivity of these two CYP154C4 proteins, crystal structures of the ligand-unbound form of StCYP154C4-1 and the testosterone-bound form of StCYP154C4-2 were determined. Comparison with the previously determined NfCYP154C5 structure revealed differences in the substrate-binding residues, suggesting a likely explanation for the different patterns of testosterone hydroxylation, despite the high sequence similarities between the enzymes (54% identity). These findings provide valuable insights that will enable protein engineering for the development of artificial steroid-related CYPs exhibiting different regiospecificity.

PubMed: 30552795
DOI: 10.1111/febs.14729

Experimental method

X-RAY DIFFRACTION (2.19 Å)