Skip to main content
Log in

Molecular analysis of methane monooxygenase from Methylococcus capsulatus (Bath)

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Methane monooxygenase (MMO) is the enzyme responsible for the conversion of methane to methanol in methanotrophic bacteria. In addition, this enzyme complex oxidizes a wide range of aliphatic and aromatic compounds in a number of potentially useful biotransformations. In this study, we have used biochemical data obtained from purification and characterization of the soluble MMO from Methylococcus capsulatus (Bath), to identify structural genes encoding this enzyme by oligonucleotide probing. The genes encoding the β and γ subunits of MMO were found to be chromosomally located and were linked in this organism. We report here on the analysis of a recombinant plasmid containing 12 kilobases of Methylococcus DNA and provide the first evidence for the localization and linkage of genes encoding the methane monooxygenase enzyme complex. DNA sequence analysis suggests that the primary structures of the β and γ subunit of MMO are completely novel and the complete sequence of these genes is presented.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Al-Taho NM, Warner PJ (1987) Restoration of phenotype in Escherichia coli auxotrophs by pULB113-mediated mobilization from methylotrophic bacteria. FEMS Microbiol Lett 43:235–239

    Google Scholar 

  • Colby J, Dalton H (1976) Some properties of a soluble methane monooxygenase from Methylococcus capsulatus (Bath). Biochem J 157:495–497

    Google Scholar 

  • Colby J, Dalton H (1978) Resolution of the methane monooxygenase of Methylococcus capsulatus (Bath) into three components: purification and properties of component C, a flavoprotein. Biochem J 171:461–468

    Google Scholar 

  • Colby J, Stirling DI, Dalton H (1977) The soluble methane monooxygenase of Methylococcus capsulatus (Bath): its ability to oxygenate n-alkanes, n-alkenes, ethers and alicylic aromatic and heterocyclic compounds. Biochem J 165:395–402

    Google Scholar 

  • Collins JF, Coulson AFW (1987) Molecular sequence comparison and alignment. In: Bischop MJ, Rawkins CJ (eds) Nucleic acid and protein sequence analysis: A practical approach. IRL Press, Oxford, pp 323–358

    Google Scholar 

  • Dalton H (1980a) Oxidation of hydrocarbons by methane monooxygenases from a variety of microbes. Adv Appl Micro 26:71–87

    Google Scholar 

  • Dalton H (1980b) Transformations by methane monooxygenase. In: Harrison DEF, Higgins, IJ, Watkinson R (eds) Hydrocarbons in biotechnology. Heyden, London, pp 85–97

    Google Scholar 

  • Dalton H, Prior SD, Leak DJ, Stanley S (1984) Regulation and control of methane monooxygenase. In: Crawford RL, Hanson RS (eds) Microbial growth on C1 compounds. Proc. 4th Inter. Symp. Am Soc Micro, Washington DC, pp 75–82

  • Deininger PL (1983) Random sub-cloning of sonicated DNA: Application to shotgun DNA sequence analysis. Anal Biochem 129:216–223

    Google Scholar 

  • Denhardt DT (1966) A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun 23:641–646

    Google Scholar 

  • Ericson A, Hedman B, Hodgson KO, Green J, Dalton H, Bentsen JG, Beer RH, Lippard SJ (1988) Structural characterization by EXAFS spectroscopy of the binuclear iron centre in protein A of methane monooxygenase from Methylococcus capsulatus (Bath). J Am Chem Soc 110:2330–2332

    Google Scholar 

  • Green J, Dalton H (1985) Protein B of soluble methane monooxygenase from Methylococcus capsulatus (Bath): a novel regulatory protein of enzyme activity. J Biol Chem 260:15795–15801

    Google Scholar 

  • Green J, Dalton H (1986) Steady-state kinetic analysis of soluble methane monooxygenase from Methylococcus capsulatus (Bath). Biochem J 236:155–162

    Google Scholar 

  • Grunstein M, Hogness DS (1975) Colony hybridization; A method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72:3961–3965

    Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Google Scholar 

  • Harwood JJ, Williams E, Bainbridge BW (1972) Mutation of the methane oxidizing bacterium Methylococcus capsulatus. J Appl Bacteriol 35:99–108

    Google Scholar 

  • Lathe R (1985) Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol 183:1–12

    Google Scholar 

  • Laursen RA (1971) Solid-phase Edman degradation. An automatic peptide sequencer. Eur J Biochem 20:89–102

    Google Scholar 

  • Lund J, Dalton H (1985) Further characterisation of the FAD and Fe2S2 redox centres of component C, the NADH: acceptor reductase of the soluble methane monooxygenase of Methylococcus capsulatus (Bath). Eur J Biochem 147:291–296

    Google Scholar 

  • Lund J, Woodland MP, Dalton H (1985) Electron transfer reactions in the soluble methane monooxygenase of Methylococcus capsulatus (Bath). Eur J Biochem 147:297–305

    Google Scholar 

  • McPheat WL, Mann NH, Dalton H (1987a) Transfer of broad host range plasmids to the type 1 obligate methanotroph Methylomonas albus. FEMS Microbiol Lett 41:185–188

    Google Scholar 

  • McPheat WL, Mann NH, Dalton H (1987b) Isolation of mutants of the obligate methanotroph Methylococcus albus defective in growth on methane. Arch Microbiol 148:40–43

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, A laboratory manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either strand of a double-digest restriction fragment. Gene 19:269–276

    Google Scholar 

  • Mullens IA, Dalton H (1987) Cloning of the gamma-subunit methane monooxygenase from Methylococcus capsulatus. Biotechnology 5:490–493

    Google Scholar 

  • Murrell JC, Dalton H (1983) Purification and properties of glutamine synthetase from Methylococcus capsulatus (Bath). J Gen Micro 129:1187–1196

    Google Scholar 

  • Nicolaidis AA, Sargent AW (1987) Isolation of methane monooxygenase deficient mutants from Methylosinus trichosporium OB3b using dichloromethane. FEMS Microbiol Lett 41:47–52

    Google Scholar 

  • Oakley CJ, Murrell JC (1988) nifH genes in the obligate methane oxidizing bacteria. FEMS Microbiol Lett 49:53–57

    Google Scholar 

  • Prior SD, Dalton H (1985) The effect of copper ions on membrane content and methane monooxygenase activity in methanol-grown cells of Methylococcus capsulatus (Bath). J Gen Microbiol 131:155–163

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Stanley SH, Prior SD, Leak DJ, Dalton H (1983) Copper stress underlies the fundamental change in intracellular location of methane monooxygenase in methane-oxidizing organisms: studies in batch and continuous cultures. Biotechnol Lett 5:487–492

    Google Scholar 

  • Stirling DI, Dalton H (1979) The fortuitous oxidation and cometabolism of various carbon compounds by whole-cell suspensions of Methylococcus capsulatus (Bath). FEMS Microbiol Lett 5:315–318

    Google Scholar 

  • Tinoco I, Borer PN, Dengler B, Levine MD, Uhlenbeck OC, Crothers DM, Gralla J (1973) Improved estimation of secondary structure in ribonucleic acids. Nature (New Biology) 246:40–41

    Google Scholar 

  • Whittenbury R, Dalton H (1981) The methylotrophic bacteria. In: Starr MP, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 894–902

    Google Scholar 

  • Williams E, Bainbridge BW (1971) Genetic transformation in Methylococcus capsulatus. J Appl Bacteriol 34:683–687

    Google Scholar 

  • Williams E, Shimmin MA, Bainbridge BW (1977) Mutation in the obligate methylotrophs Methylococcus capsulatus and Methylomonas albus. FEMS Microbiol Lett 2:293–296

    Google Scholar 

  • Woodland MP, Dalton H (1984a) Purification of component A of the soluble methane monooxygenase of Methylococcus capsulatus (Bath) by high-pressure gel permeation chromatography. Anal Biochem 139:459–462

    Google Scholar 

  • Woodland MP, Dalton H (1984b) Purification and properties of component A of the methane monooxygenase from Methylococcus capsulatus (Bath). J Biol Chem 259:53–59

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stainthorpe, A.C., Murrell, J.C., Salmond, G.P.C. et al. Molecular analysis of methane monooxygenase from Methylococcus capsulatus (Bath). Arch. Microbiol. 152, 154–159 (1989). https://doi.org/10.1007/BF00456094

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00456094

Key words

Navigation