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