The steroid 15β-hydroxylase system of Bacillus megaterium was obtained in a cell-free preparation through sonication. The strictly NADPH-dependent 15β-hydroxylase activity, measured using progesterone as substrate, was inhibited by carbon monoxide, SKF 525-A, imidazole and metyrapone, indicating that the reaction is cytochrome P-450-dependent. A 40-fold purification of cytochrome P-450 in cell-free extracts was obtained by chromatography on DEAE-cellulose yielding a concentration of 0.32 nmoles of cytochrome P-450 per mg of protein. This partially purified cytochrome P-450 preparation catalyzed 15β- and 15α-hydroxylation of progesterone in the presence of NaIO4 or NaClO2 but not in the presence of NADPH or NADH.
|Original language||English (US)|
|Number of pages||10|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Oct 27 1975|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology