title: In vitro analysis of polypeptide requirements of multicomponent phenol hydroxylase from Pseudomonas sp. strain CF600.
abstract: An in vitro study of the multicomponent phenol hydroxylase from Pseudomonas sp. strain CF600 was performed. Phenol-stimulated oxygen uptake from crude extracts was strictly dependent on the addition of NAD(P)H and Fe2+ to assay mixtures. Five of six polypeptides required for growth on phenol were necessary for in vitro activity. One of the polypeptides was purified to homogeneity and found to be a flavin adenine dinucleotide containing iron-sulfur protein with significant sequence homology, at the amino terminus, to plant-type ferredoxins. This component, as in other oxygenase systems, probably functions to transfer electrons from NAD(P)H to the iron-requiring oxygenase component. Phenol hydroxylase from this organism is thus markedly different from bacterial flavoprotein monooxygenases commonly used for hydroxylation of other phenolic compounds, but bears a number of similarities to multicomponent oxygenase systems for unactivated compounds.
authors: Unit for Applied Cell and Molecular Biology, University of Umeå, Sweden.
authors: J
authors: J
authors: Powlowski
authors: V
authors: V
authors: Shingler
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D046911:Macromolecular Substances; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism; D019800:Phenol; D010636:Phenols / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology*; D013329:Structure-Activity Relationship
chemical_list: D046911:Macromolecular Substances; D010636:Phenols; D019800:Phenol; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C022336:phenol 2-monooxygenase; D009245:NADH Dehydrogenase