Camphor revisited: studies of 2,5-diketocamphane 1,2-monooxygenase from Pseudomonas putida ATCC 17453.

摘要: The oxygenating component of 2,5-diketocamphane 1,2-monooxygenase from Pseudomonas putida ATCC 17453 was purified to homogeneity by a combination ammonium sulfate fractionation and chromatography on DEAE-cellulose polyanion SI-17 columns. It had an Mr 78,000, bound one molecule nonautooxidizable flavin mononucleotide (FMN), consisted two subunits equal molecular weight, existed in electrophoretically distinguishable active forms. complex constructed equimolecular amounts NADH oxidase, which could be separately (Mr, 36,000), the component. Most oxidase dissociated during purification, although traces remained, give final preparation significant oxygenase activity. FMN did not dissociate significantly but it covalently removed under variety conditions. Binding between proteins that made up fairly weak freely reversible. probably occurred through strongly for binding site. Iron present at level component, common with other characterized Baeyer Villiger monooxygenases, found simple flavoprotein.