The inducible and cytochrome P450-containing dehydroepiandrosterone 7alpha-hydroxylating enzyme system of Fusarium moniliforme

J Steroid Biochem Mol Biol. 1997 Aug;62(5-6):467-75. doi: 10.1016/s0960-0760(97)00060-5.

Abstract

7Alpha-hydroxylation of DHEA by Fusarium moniliforme was investigated with regard to inducibility and characterization of the responsible enzyme system. Using GC/MS, the 7-hydroxylated metabolites of DHEA produced after biotransformation by Fusarium moniliforme mycelia were identified. The strain of Fusarium moniliforme hydroxylated DHEA predominantly at the 7alpha-position, with minor hydroxylation occurring at the 7beta-position. Constitutive 7alpha-hydroxylation activity was low, but DHEA induced the enzyme complex responsible for 7alpha-hydroxylation via an increase in protein synthesis. DHEA 7alpha-hydroxylase was found to be mainly microsomal, and the best production yields of 7alpha-hydroxy-DHEA (28.5 +/- 3.51 pmol/min/mg protein) were obtained with microsomes prepared from 18-h-induced mycelia. Kinetic parameters (KM=1.18 +/- 0.035 microM and Vmax=909 +/- 27 pmol/min/mg protein) were determined. Carbon monoxide inhibited 7alpha-hydroxylation of DHEA by microsomes of Fusarium moniliforme. Also, exposure of mycelia to DHEA increased microsomal P450 content. These results demonstrated that: (i) DHEA is 7alpha-hydroxylated by microsomes of Fusarium moniliforme; (ii) DHEA induces Fusarium moniliforme 7alpha-hydroxylase; (iii) this enzyme complex contains a cytochrome P450.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Monoxide / pharmacology
  • Cytochrome P-450 Enzyme System / metabolism*
  • Dehydroepiandrosterone / analogs & derivatives
  • Dehydroepiandrosterone / metabolism*
  • Enzyme Induction
  • Fusarium / drug effects
  • Fusarium / enzymology*
  • Fusarium / metabolism
  • Hydroxylation
  • Kinetics
  • Microsomes / enzymology
  • Steroid Hydroxylases / biosynthesis
  • Steroid Hydroxylases / metabolism*

Substances

  • Dehydroepiandrosterone
  • 7-hydroxydehydroepiandrosterone
  • Carbon Monoxide
  • Cytochrome P-450 Enzyme System
  • Steroid Hydroxylases
  • oxysterol 7-alpha-hydroxylase