title: Purification, crystallisation and characterization of quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa.
abstract: Pseudomonas aeruginosa ATCC 17933 when grown on ethanol produces high levels of a quinoprotein ethanol dehydrogenase, which amounts to 7% of the soluble protein. The enzyme has been purified to homogeneity and it crystallizes readily in the presence of polyethylene glycol 1550 or 6000. The ethanol dehydrogenase (Km(ethanol) = 14 microM) resembles the dye-dependent quinoprotein methanol dehydrogenases of methylotrophic bacteria, but has a low affinity for methanol (Km (methanol) = 94mM). In addition the enzyme oxidizes secondary alcohols. With its catalytic properties the ethanol dehydrogenase is similar to the enzyme isolated from P. aeruginosa LMD 80.53 (Groen, B., Frank, J. Jzn. & Duine, J.A. (1984) Biochem. J. 223, 921-924). In contrast to this enzyme from P. aeruginosa LMD 80.53, which is a monomer, the ethanol dehydrogenase isolated from P. aeruginosa ATCC 17933 is a dimer of identical subunits of relative molecular mass 60,000. The N-terminal amino acid is lysine. Inactivation with cyclopropanone ethylhemiketal reveals one molecule of pyrroloquinoline quinone per subunit. As shown by active enzyme sedimentation, the dimer is the enzymatically active form.
authors.affiliation: Institut für Mikrobiologie der Universität Hohenheim.
authors.forename: M
authors.initials: M
authors.lastname: Rupp
authors.forename: H
authors.initials: H
authors.lastname: Görisch
mesh_terms: D000429:Alcohol Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011549:Pseudomonas / Q000201:enzymology; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D046911:Macromolecular Substances; D000429:Alcohol Oxidoreductases; C019641:alcohol dehydrogenase (acceptor)