Alcohol dehydrogenase from Methylobacterium organophilum.
The alcohol dehydrogenase from Methylobacterium organophilum, a facultative methane-oxidizing bacterium, has been purified to homogeneity as indicated by sodium dodecyl sulfate-gel electrophoresis. It has several properties in common with the alcohol dehydrogenases from other methylotrophic bacteria. The active enzyme is a dimeric protein, both subunits having molecular weights of about 62,000. The enzyme exhibits broad substrate specificity for primary alcohols and catalyzes the two-step oxidation of methanol to formate. The apparent Michaelis constants of the enzyme are 2.9 x 10(-5) M for methanol and 8.2 x 10(-5) M for formaldehyde. Activity of the purified enzyme is dependent on phenazine methosulfate. Certain characteristics of this enzyme distinguish it from the other alcohol dehydrogenases of other methylotrophic bacteria. Ammonia is not required for, but stimulates the activity of newly purified enzyme. An absolute dependence on ammonia develops after storage of the purified enzyme. Activity is not inhibited by phosphate. The fluorescence spectrum of the enzyme indicates that it and the cofactor associated with it may be chemically different from the alcohol dehydrogenases from other methylotrophic bacteria. The alcohol dehydrogenases of Hyphomicrobium WC-65, Pseudomonas methanica, Methylosinus trichosporium, and several facultative methylotrophs are serologically related to the enzyme purified in this study. The enzymes of Rhodopseudomonas acidophila and of organisms of the Methylococcus group did not cross-react with the antiserum prepared against the alcohol dehydrogenase of M. organophilum.
D000429:Alcohol Oxidoreductases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000643:Ammonium Chloride / Q000494:pharmacology; D002474:Cell-Free System; D000939:Epitopes; D005557:Formaldehyde / Q000378:metabolism; D005561:Formates / Q000096:biosynthesis; D008697:Methane / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D008772:Methylococcaceae / Q000201:enzymology* / Q000276:immunology; D008970:Molecular Weight; D010084:Oxidation-Reduction; D013379:Substrate Specificity
D000939:Epitopes; D005561:Formates; D000643:Ammonium Chloride; D005557:Formaldehyde; D000429:Alcohol Oxidoreductases; D008697:Methane; D000432:Methanol