title: Flavocytochrome P450 BM3: an update on structure and mechanism of a biotechnologically important enzyme.
abstract: Since its discovery in the 1980s, the fatty acid hydroxylase flavocytochrome P450 (cytochrome P450) BM3 (CYP102A1) from Bacillus megaterium has been adopted as a paradigm for the understanding of structure and mechanism in the P450 superfamily of enzymes. P450 BM3 was the first P450 discovered as a fusion to its redox partner--a eukaryotic-like diflavin reductase. This fact fuelled the interest in soluble P450 BM3 as a model for the mammalian hepatic P450 enzymes, which operate a similar electron transport chain using separate, membrane-embedded P450 and reductase enzymes. Structures of each of the component domains of P450 BM3 have now been resolved and detailed protein engineering and molecular enzymology studies have established roles for several amino acids in, e.g. substrate binding, coenzyme selectivity and catalysis. The potential of P450 BM3 for biotechnological applications has also been recognized, with variants capable of industrially important transformations generated using rational mutagenesis and forced evolution techniques. This paper focuses on recent developments in our understanding of structure and mechanism of this important enzyme and highlights important problems still to be resolved.
authors.affiliation: Department of Biochemistry, University of Leicester, The Henry Wellcome Building, Lancaster Road, Leicester LE1 9HN, UK. awm9@le.ac.uk
authors.forename: A J
authors.initials: AJ
authors.lastname: Warman
authors.forename: O
authors.initials: O
authors.lastname: Roitel
authors.forename: R
authors.initials: R
authors.lastname: Neeli
authors.forename: H M
authors.initials: HM
authors.lastname: Girvan
authors.forename: H E
authors.initials: HE
authors.lastname: Seward
authors.forename: S A
authors.initials: SA
authors.lastname: Murray
authors.forename: K J
authors.initials: KJ
authors.lastname: McLean
authors.forename: M G
authors.initials: MG
authors.lastname: Joyce
authors.forename: H
authors.initials: H
authors.lastname: Toogood
authors.forename: R A
authors.initials: RA
authors.lastname: Holt
authors.forename: D
authors.initials: D
authors.lastname: Leys
authors.forename: N S
authors.initials: NS
authors.lastname: Scrutton
authors.forename: A W
authors.initials: AW
authors.lastname: Munro
mesh_terms: D000818:Animals; D001410:Bacillus megaterium / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D008322:Mammals; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D009251:NADPH-Ferrihemoprotein Reductase; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D051381:Rats
chemical_list: D001426:Bacterial Proteins; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases