title: Involvement of cytochrome P-450 in the 15alpha-hydroxylation of 13-ethyl-gon-4-ene-3,17-dione by Penicillium raistrickii.
abstract: The 15alpha-hydroxylation of 13-ethyl-gon-4-ene-3,17-dione (GD) with different subcellular fractions of Penicillium raistrickii i 477 was investigated. Cytochrome P-450 was shown to be involved in this reaction. The steroid transformation was inhibited by carbon monoxide, metyrapone, p-CMB, iodoacetamide, N-methylmaleimide and several metal ions. The 15alpha-hydroxylase was observed to be dependent on nicotinamide-adenine dinucleotide phosphate (NADPH) replaceable by NaIO4, and the activity was enhanced by a NADPH-regenerating system, indicating the involvement of the NADPH-cytochrome c (P-450) reductase. This was further confirmed by the inhibition of the hydroxylase activity in the presence of cytochrome c. No effect was observed in the presence of azide and antimycin A. Solubilized microsomes gave an absorption maximum at 453 nm in carbon monoxide difference spectrum, and showed a Type-I GD-binding spectrum typically for cytochrome P-450 interaction with substrate. First results about the inducibility of the enzymes involved in the 15alpha-hydroxylation of GD are shown.
authors.affiliation: Friedrich-Schiller-Universität Jena, Biologisch-Pharmazeutische Fakultät, Institut für Mikrobiologie, Lehrstuhl Technische Mikrobiologie, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Irrgang
authors.forename: D
authors.initials: D
authors.lastname: Schlosser
authors.forename: W
authors.initials: W
authors.lastname: Fritsche
mesh_terms: D002248:Carbon Monoxide / Q000494:pharmacology; D002451:Cell Compartmentation; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004790:Enzyme Induction; D004791:Enzyme Inhibitors; D006067:Gonanes / Q000378:metabolism*; D006900:Hydroxylation; D008861:Microsomes / Q000201:enzymology; D010407:Penicillium / Q000201:enzymology*; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: C107076:13-ethyl-15-hydroxygon-4-ene-3,17-dione; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D006067:Gonanes; C055308:13-ethyl-gon-4-ene-3,17-dione; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System