The distribution of methane mono-oxygenase (MMO) activities between particulate and soluble fractions of cell-free extracts of Methylosinus trichosporium OB3b was dependent upon growth conditions. Particulate activity was associated with the presence of intracytoplasmic membranes observed only under oxygen-limiting conditions in shake flask cultures. Particulate and soluble activities showed substantially different sensitivities to a range of potential inhibitors. The particulate enzyme was inhibited by metal-chelating agents, thiol reagents and amytal, whereas the soluble MMO was not inhibited by these compounds; both activities were sensitive to KCN, ethyne and 8-hydroxyquinoline. NAD(P)H was the only suitable electron donor. The activities were unstable at 0° C but the soluble enzyme could be partially stabilized by several compounds. The particulate and soluble MMO activities are compared with previously reported particulate and soluble MMO enzymes from this species and other methane oxidizers.
BestD. J.,
HigginsI. J.1981; Methane-oxidizing activity and membrane morphology in a methanol-grown obligate methanotroph, Methylosinus trichosporium OB3b. Journal of General Microbiology 125:73–84
BrannanJ.,
HigginsI. J.1978; Effect of growth conditions on the intracytoplasmic membranes of Methylosinus trichosporium OB3b. Proceedings of the Society for General Microbiology 5:69
ColbyJ.,
DaltonH.,
WhittenburyR.1975; An improved assay for bacterial methane monooxygenase: some properties of the enzyme from Methylomonas methanica. Biochemical Journal 151:459–462
ColbyJ.,
StirlingD. I.,
DaltonH.1977; The soluble methane mono-oxygenase of Methylococcus capsulatus (Bath).Its ability to oxygenate n-alkanes, n-alkenes, ethers, and alicyclic, aromatic and heterocyclic compounds. Biochemical Journal 165:395–402
GornallA. G.,
BardawillC. J.,
DavidM. M.1949; Determination of serum proteins by means of the biuret reaction. Journal of Biological Chemistry 177:751–766
HammondR. C.,
TaylorF.,
HigginsI. J.1979; Studies on the mechanism of methane mono-oxygenase of Methylosinus trichosporium OB3b. Society for General Microbiology Quarterly 6:89
HigginsI. J.,
KnowlesC. J.,
TongeG. M.1977; Enzymic mechanisms of methane and methanol oxidation in relation to electron transport systems in methylotrophs; purification and proper-ties of methane oxygenase. In Microbial Production and Utilization of Gases (H2, CH4, CO) pp. 389–402SchlegelH. G.
Edited by Gottingen: GoltzeDruck.;
HigginsI. J.,
BestD. J.,
ScottD.1981; Hydrocarbon oxidation by Methylosinus trichosporium: metabolic implications of the lack of specificity of methane mono-oxygenase. In Proceedings of the 3rd International Symposium on Microbial Growth on C1 Compounds pp. 11–20DaltonH.
Edited by London: Heyden.;
HyderS. L.,
MeyersA.,
CayerM. L.1979; Membrane modulation in a methylotrophic bacterium Methylococcus capsulatus (Texas) as a function of growth substrate. Tissue and Cell 11:597–610
KellenbergerE.,
RyterA.,
SechaudJ.1958; Electron microscope study of DNA-containing plasms. II. Vegetative and mature phage DNA as compared with normal bacterial nucleoids in different physiological states. Journal of Biophysical and Biochemical Cytology 4:671–678
PatelR. N.,
HouC. T.,
LaskinA. I.,
FelixA.,
DerelankoP.1979; Microbial oxidation of gaseous hydrocarbons. II. Hydroxylation of alkanes and epoxidation of alkenes by cell-free particulate fractions of methane-utilizing bacteria. Journal of Bacteriology 139:675–679
PattT. E.,
HansonR. S.1978; Intracytoplasmic membrane, phospholipid, and sterol content of Methylobacterium organophilum cells grown under different conditions. Journal of Bacteriology 134:636–644
RibbonsD. W.1975; Oxidation of C1 compounds by particulate fractions from Methylococcus capsulatus: distribution and properties of methanedependent reduced nicotinamide adenine dinucleotide oxidase (methane hydroxylase). Journal of Bacteriology 122:1351–1363
StirlingD. I.,
DaltonH.1977; Effect of metal-binding agents and other compounds on methane oxidation by two strains of Methylococcus capsulatus. Archives of Microbiology 114:71–76
StirlingD. I.,
DaltonH.1979; Properties of the methane mono-oxygenase from extracts of Methylosinus trichosporium OB3b and evidence for its similarity to the enzyme from Methylococcus capsulatus (Bath). European Journal of Biochemistry 96:205–212
TongeG. M.,
HarrisonD. E. F.,
KnowlesC. J.,
HigginsI. J.1975; Properties and partial purification of the methane-oxidizing enzyme system from Methylosinus trichosporium. FEBS Letters 58:293–299
TongeG. M.,
HarrisonD. E. F.,
HigginsI. J.1977; Purification and properties of the methane mono-oxygenase enzyme system from Methylosinus trichosporium OB3b. Biochemical Journal 161:333–344
WhittenburyR.,
PhillipsK. C.,
WilkinsonJ. F.1970; Enrichment, isolation and some properties of methane-utilizing bacteria. Journal of General Microbiology 61:205–218
WolfeR. S.,
HigginsI. J.1979; Microbial biochemistry of methane - a study in contrasts. In International Review of Biochemistry21 pp. 267–353QuayleJ. R.
Edited by Baltimore: University Park Press.;