Mechanistic studies on a cytochrome P450 enzyme from the bacteria Bacillus megaterium

A cytochrome P450 enzyme from the bacteria Bacillus megaterium (P450BM-3) has been shown to have unique structural and functional properties. P450BM-3 strikingly contrasts with other P450 enzymes as it is a soluble, catalytically self-sufficient, single polypeptide protein, with both electron transfer and substrate oxidation domains contained in one protein. For these results, possibly, P450BM-3 is the most catalytically active P450 enzyme known to date.;P450BM-3 catalyses the hydroxylation of long to mid-chain fatty acids, and fatty amides and alcohols to corresponding monohydroxy derivatives on the -1, -2 and -3 positions. The potential application of P450BM-3 in synthetic chemistry has been assessed in studies using a range of synthesised long chain fatty acid derivatives.;Chapter 2 describes a study of 13(R)-hydroxymyristic acid, 12(R)-hydroxymyristic acid and 12(S)-hydroxymyristic acid as substrates, which has uncovered a degree of diastereoselectivity in the formation of 12,13-dihydroxymyristic acid products. In chapter 3 a range of mid-chain phenoxy compounds, with varying lengths of terminal alkyl chains, has been used to show regioselectivity in the P450BM-3 oxidation of these types of substrates to hydroxy and keto derivatives. In chapter 4, 11-phenylsulphanylundecanoic acid has been shown to be oxidised by P450BM-3 to a corresponding sulphoxide with a high degree of regioselectivity but non-stereoselectively.;The synthesis of substrates and the characterisation of products from corresponding P450BM-3 metabolisms are fully discussed.