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Characterization of two steroid hydroxylases from different Streptomyces spp. and their ligand­bound and ­unbound crystal structures

Cited 3 time in wos
Cited 3 time in scopus
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Title
Characterization of two steroid hydroxylases from different Streptomyces spp. and their ligand­bound and ­unbound crystal structures
Other Titles
두 가지 다른 방선균으로부터의 스테로이드 수산화 효소의 생화학적 특성 및 삼차구조 연구
Authors
Dangi, Bikash
Lee, Chang Woo
Kim, Ki-Hwa
Park, Sun-Ha
Yu, Eun-Ji
Jeong, Chang-Sook
Park, Hyun
Lee, Jun Hyuck
Oh, Tae-Jin
Subject
Biochemistry; Molecular Biology
Keywords
Streptomyces; X-ray crystallography; crystal structure; cytochrome P450; steroid hydroxylase
Issue Date
2019-05
Citation
Bikash Dangi, et al. 2019. "Characterization of two steroid hydroxylases from different Streptomyces spp. and their ligand­bound and ­unbound crystal structures". FEBS JOURNAL, 286(9): 1683-1699.
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, two 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.
URI
https://repository.kopri.re.kr/handle/201206/10403
DOI
http://dx.doi.org/10.1111/febs.14729
ISSN
1742-464X
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