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Characterization of CYP125A13, the First Steroid C-27 Monooxygenase from Streptomyces peucetius ATCC27952

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Title
Characterization of CYP125A13, the First Steroid C-27 Monooxygenase from Streptomyces peucetius ATCC27952
Other Titles
항생물질 생산 방선균인 Streptomyces peucetius 균주유래 스테로이드 C-27번 위치에 산소원자를 붙이는 CYP125A13 효소의 모델링 구조분석 및 생화학적 특성 분석
Authors
Rimal, Hemraj
Subedi, Pradeep
Kim, Ki-Hwa
Park, Hyun
Lee, Jun Hyuck
Oh, Tae-Jin
Subject
Biotechnology & Applied Microbiology; Microbiology
Keywords
Streptomyces peucetius; cytochrome P450; CYP125A13; 27-hydroxycholesterol; regio-selective hydroxylation
Issue Date
2020-11
Citation
Rimal, Hemraj, et al. 2020-11. "Characterization of CYP125A13, the First Steroid C-27 Monooxygenase from Streptomyces peucetius ATCC27952". JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, 30(11): 1750-1759.
Abstract
The characterization of cytochrome P450 CYP125A13 from Streptomyces peucetius was conducted using cholesterol as the sole substrate. The in vitro enzymatic assay utilizing putidaredoxin and putidaredoxin reductase from Pseudomonas putida revealed that CYP125A13 bound cholesterol and hydroxylated it. The calculated K-D value, catalytic conversion rates, and Km value were 56.92 +/- 11.28 mu M, 1.95 nmol min(-1) nmol(-1), and 11.3 +/- 2.8 mu M, respectively. Gas chromatography-mass spectrometry (GC-MS) analysis showed that carbon 27 of the cholesterol side-chain was hydroxylated, characterizing CYP125A13 as steroid C27-hydroxylase. The homology modeling and docking results also revealed the binding of cholesterol to the active site, facilitated by the hydrophobic amino acids and position of the C27-methyl group near heme. This orientation was favorable for the hydroxylation of the C27-methyl group, supporting the in vitro analysis. This was the first reported case of the hydroxylation of cholesterol at the C-27 position by Streptomyces P450. This study also established the catalytic function of CYP125A13 and provides a solid basis for further studies related to the catabolic potential of Streptomyces species.
URI
https://repository.kopri.re.kr/handle/201206/11867
DOI
http://dx.doi.org/10.4014/jmb.2007.07004
Appears in Collections  
2020-2020, Development of potential candidates as antibiotics based on polar genetic resources (20-20) / Lee, Jun Hyuck (PM20030)
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