Oxidation of C1 compounds by particulate fractions from Methylococcus capsulatus: properties of methanol oxidase and methanol dehydrogenase.
Methanol (and formaldehyde) oxidizing activities in crude extracts of Methylococcus capsulatus are associated mainly with particulate fractions sedimenting between 3,000 and 40,000 X g. Most of the phenazine methosulfate (PMS)-dependent methanol (and formaldehyde) dehydrogenase activity observed resides in the soluble fraction but represents only 40% of the total (PMS dependent plus independent) activity. Both PMS-dependent methanol dehydrogenase activity and PMS-independent methanol oxidase activity are found in particulate fractions, and the PMS-dependent dehydrogenase is easily solubilized by treatment with certain phospholipases or detergents. The properties of the PMS-dependent dehydrogenase activities in the soluble fraction and that solubilized from the particles suggested that they may be identical proteins. Their pH optima, temperature dependence, thermolabilities, and sensitivities to the presence of specific antisera were indistinguishable. Homogeneous preparations of the enzyme proteins obtained from the soluble fractions of extracts and the particulate fractions solubilized by detergents had similar: (i) electrophoretic mobilities in native and denatured states (subunit size in sodium dodecyl sulfate  62,000 daltons); (ii) molecular radii under native conditions, (iii) visible absorption spectra, lambdamax 350 nm, (iv) kinetic constants for methanol and formaldehyde; (v) substrate specificity; and (vi) immunological characteristics--antisera to each enzyme preparation showed precipitin lines of identity to either of the enzymes. It is suggested that the major site of methanol and formaldehyde oxidation in M. capsulatus occurs on the intracytoplasmic membranes in vivo and is coupled to oxygen reduction.
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D004589:Electrophoresis, Disc; D005557:Formaldehyde / Q000378:metabolism*; D005561:Formates / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D000432:Methanol / Q000378:metabolism*; D008773:Methylphenazonium Methosulfate / Q000378:metabolism; D008835:Micrococcaceae / Q000201:enzymology* / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism*; D010740:Phospholipases; D012995:Solubility; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism; D013501:Surface-Active Agents; D013696:Temperature
D005561:Formates; D013501:Surface-Active Agents; D005557:Formaldehyde; D008773:Methylphenazonium Methosulfate; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; D010740:Phospholipases; D000432:Methanol