Flavin supported fatty acid oxidation by the heme domain of Bacillus megaterium cytochrome P450BM-3

Biochem Biophys Res Commun. 1994 Sep 30;203(3):1745-9. doi: 10.1006/bbrc.1994.2388.

Abstract

Cytochrome P450BM-3 is a fatty acid hydroxylase that consists of a heme domain covalently attached to a diflavin (FMN+FAD) cytochrome P450 reductase domain. The heme and flavin domains can be separately expressed and purified from E. coli recombinant expression systems. Normally P450s require a protein redox partner as a source of electrons. We now have found that the P450BM-3 heme domain can be reduced by NADPH+FMN and that reduced FMN can support the P450 catalyzed hydroxylation of a fatty acid substrate, myristic acid. HPLC profiles show that the "artificial" FMN supported hydroxylation gives the same products as does holo-P450BM-3.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacillus megaterium / enzymology*
  • Bacterial Proteins*
  • Carbon Radioisotopes
  • Chromatography, High Pressure Liquid
  • Cytochrome P-450 Enzyme System / metabolism*
  • Flavin Mononucleotide / metabolism*
  • Flavin Mononucleotide / pharmacology
  • Heme / metabolism
  • Mixed Function Oxygenases / metabolism*
  • Myristic Acid
  • Myristic Acids / metabolism*
  • NADP / metabolism
  • NADPH-Ferrihemoprotein Reductase
  • Oxidation-Reduction
  • Radioisotope Dilution Technique
  • Spectrophotometry

Substances

  • Bacterial Proteins
  • Carbon Radioisotopes
  • Myristic Acids
  • Myristic Acid
  • Heme
  • NADP
  • Flavin Mononucleotide
  • Cytochrome P-450 Enzyme System
  • Mixed Function Oxygenases
  • NADPH-Ferrihemoprotein Reductase
  • flavocytochrome P450 BM3 monoxygenases