Demonstration of a cytochrome P-450-dependent steroid 15β-hydroxylase in Bacillus megaterium

Anders Berg, Kjell Carlström, Jan Åke Gustafsson, Magnus Ingelman-Sundberg

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44 Scopus citations

Abstract

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 languageEnglish (US)
Pages (from-to)1414-1423
Number of pages10
JournalBiochemical and Biophysical Research Communications
Volume66
Issue number4
DOIs
StatePublished - Oct 27 1975

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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