Methane Estimation for Methanogenic and Methanotrophic Bacteria

  • M. R. Smith
  • L. Baresi
Part of the Modern Methods of Plant Analysis book series (MOLMETHPLANT, volume 9)


Methanogenic and methanotrophic bacteria frequently occur in the same habitats, the methanogens producing methane and the methanotrophs consuming the methane produced by the methanogens. The activities of both bacterial groups can be monitored by measuring methane in the atmospheres of cultures and ecosystem samples. However, there is a greater reliance on methane measurement for methanogenic than for methanotrophic bacteria. In this chapter, we cover methods for estimating methane and associated gases. A few specialized methods for each group will also be considered.


Methanogenic Bacterium Methanotrophic Bacterium Methane Monooxygenase Combustion Tube Anaerobic Methane Oxidation 
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  1. Ackman RG (1972) Porous polymer bead packings and formic acid vapor in the GLC of volatile free fatty acids. J Chromatogr Sci 10: 560–565PubMedGoogle Scholar
  2. Alperin MJ, Reeburgh WS (1985) Inhibition experiments on anaerobic methane oxidation. Appl Environ Microbiol 50: 940–945PubMedGoogle Scholar
  3. Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens:reevaluation of a unique biological group. Microbiol Rev 43: 260–296PubMedGoogle Scholar
  4. Baresi L (1984) Methanogenic cleavage of acetate by lysates of Methanosarcina sp. J Bac-teriol 160: 365–370Google Scholar
  5. Baresi L, Mah RA, Ward DM, Kaplan IR (1978) Methanogenesis from acetate:enrichment studies. Appl Environ Microbiol 36: 186–197PubMedGoogle Scholar
  6. Belay N, Daniels L (1987) Production of ethane, ethylene, and acetylene from halogenated hydrocarbons by methanogenic bacteria. Appl Environ Microbiol 53: 1604–1610PubMedGoogle Scholar
  7. Ben-Nairn A (1972) Thermodynamics of dilute aqueous solutions of nonpolar solutes. In:Horne RA (ed) Water and aqueous solutions structure, thermodynamics, and transport processes. Wiley, New York London Sydney Toronto, p 430Google Scholar
  8. Best DJ, Higgins IJ (1981) Methane-oxidizing activity and membrane morphology in a methanol-grown obligate methanotroph, Methylosinus trichosporium OB3b. J Gen Microbiol 125: 73–84Google Scholar
  9. Blaut M, Gottschalk G (1982) Effect of trimethylamine on acetate utilization by Methanosarcina barken. Arch Microbiol 133: 230–235CrossRefGoogle Scholar
  10. Breck DW (1964) Crystalline molecular sieves. J Chem Ed 41: 678–685CrossRefGoogle Scholar
  11. Brewer JM, Pesce AJ, Ashworth RB (eds) (1974) Experimental techniques in biochemistry. Prentice-Hall, Englewood Cliffs, NJ, pp 374Google Scholar
  12. Bulletin 712B (1976) Carbosieve brand S-GSC packing. Supelco, Bellefonte, PAGoogle Scholar
  13. Bulletin 760A (1976) Analysis of permanent gases. Supelco, Bellefonte, PAGoogle Scholar
  14. Bulletin 769 (1977) Determination of organic vapors in the industrial atmosphere. Supelco, Bellefonte, PAGoogle Scholar
  15. Bulletin (1981) Carbosphere. Appl Sci Lab, Deerfield, ILGoogle Scholar
  16. Caprioli RM (1972) Use of stable isotopes. In:Waller RG (ed) Biochemical applications of mass spectrometry. Wiley, New York London Sydney Toronto, pp 735–779Google Scholar
  17. Carlsson J (1973) Simplified gas Chromatographic procedure for identification of bacterial metabolic products. Appl Microbiol 25: 287–289PubMedGoogle Scholar
  18. Cochrane GC (1975) A review of the analysis of free fatty acids [C2–C6]-J Chromatogr Sci 13: 440–447Google Scholar
  19. Colby J, Dalton H (1978) Resolution of the methane mono-oxygenase of Methylococcus capsulatus (Bath) into three components. Purification and properties of component C, a flavoprotein. Biochem J 171: 461–468Google Scholar
  20. Colby J, Dalton H, Whittenbury R (1975) An improved assay for bacterial methane mono-oxygenase:some properties of the enzyme from Methylomonas methanica. Biochem J 151: 459–462PubMedGoogle Scholar
  21. Colby J, Dalton H, Whittenbury R (1979) Biological and biochemical aspects of microbial growth on C1 compounds. Annu Rev Microbiol 33: 481–517PubMedCrossRefGoogle Scholar
  22. Conrad R, Thauer RK (1983) Carbon monoxide production by Methanobacterium thermo-autotrophicum. FEMS Microbiol Lett 20: 229–232CrossRefGoogle Scholar
  23. Cramers CA, McNair HM (1983) Gas chromatography. In:Heftmann E (ed) Chromatog-raphy fundamentals and applications of Chromatographie and electrophoretic techniques, pt A:fundamentals and techniques. Elsevier, Amsterdam Oxford New York, pp A195–A224CrossRefGoogle Scholar
  24. Dalton H (1980) Oxidation of hydrocarbons by methane monooxygenases from a variety of microbes. Adv Appl Microbiol 26: 71–87CrossRefGoogle Scholar
  25. Daniels L, Fuchs G, Thauer RK, Zeikus JG (1977) Carbon monoxide oxidation by metha-nogenic bacteria. J Bacteriol 132: 18–126Google Scholar
  26. Daniels L, Fulton G, Spencer RW, Orme-Johnson WH (1980) Origin of hydrogen in methane produced by Methanobacterium thermoautotrophicum. J Bacteriol 141: 694–698PubMedGoogle Scholar
  27. Dean JA (ed) (1973) Lange’s handbook of chemistry, 11th edn. McGraw-Hill, New York, pp 1015–1017Google Scholar
  28. Dennis LM, Nichols ML (1929) Gas analysis. MacMillan, New York, pp 138–259Google Scholar
  29. Ferenci T (1974) Carbon monoxide-stimulated respiration in methane-utilizing bacteria. FEBS Lett 41: 94–98PubMedCrossRefGoogle Scholar
  30. Ferenci T, Strom T, Quayle JR (1975) Oxidation of carbon monoxide and methane by Pseudomonas methanica. J Gen Microbiol 91: 79–91PubMedGoogle Scholar
  31. Ferry JG, Wolfe RW (1976) Anaerobic degradation of benzoate to methane by a microbial consortium. Arch Microbiol 107: 33–40PubMedCrossRefGoogle Scholar
  32. Fuchs G, Thauer R, Ziegler H, Stichler W (1979) Carbon isotope fractionation by Methanobacterium thermoautotrophicum. Arch Microbiol 120: 135–139CrossRefGoogle Scholar
  33. Glover J (1956) Methods involving labeled atoms. In:Paech K, Tracey MV (eds) Modern methods of plant analysis, vol 1. Springer, Berlin Göttingen Heidelberg, pp 325–374Google Scholar
  34. Gnaiger E, Forstner H (eds) (1983) Polarographic oxygen sensors. Springer, Berlin Heidelberg New York, pp 370Google Scholar
  35. Hanson RS (1980) Ecology and diversity of methylotrophic organisms. Adv Appl Microbiol 26: 3–39CrossRefGoogle Scholar
  36. Hash JH (1972) Liquid scintillation counting in microbiology. In:Norris JR, Ribbons RW (eds) Methods in microbiology, vol 6 B. Academic Press, New York London, pp 109–155Google Scholar
  37. Higgins IJ, Quayle JR (1970) Oxygenation of methane by methane-grown Pseudomonas methanica and Methanomonas methanooxidans. Biochem J 118: 201–208PubMedGoogle Scholar
  38. Higgins IJ, Best DJ, Hammond RC, Scott D (1981) Methane-oxidizing microorganisms. Microbiol Rev 45: 556–590PubMedGoogle Scholar
  39. Hippe H, Caspari D, Fiebig K, Gottschalk G (1979) Utilization of trimethylamine and other N-methyl compounds for growth and methane formation by Methanosarcina barken. Proc Natl Acad Sci Usa 76: 494–498PubMedCrossRefGoogle Scholar
  40. Hou CT, Patel RN, Laskin AI, Barnabe N (1979) Microbial oxidation of gaseous hydrocarbons:epoxidation of n-alkenes by methylotrophic bacteria. Appl Environ Microbiol 38: 127–134PubMedGoogle Scholar
  41. Hou CT, Patel RN, Laskin AI (1980) Epoxidation and ketone formation by C1-utilizing microbes. Adv Appl Microbiol 26: 41–69CrossRefGoogle Scholar
  42. Imai T, Takigawa H, Nakagawa S, Shen S, Kodama T, Minoda Y (1986) Microbial oxidation of hydrocarbons and related compounds by whole-cell suspensions of the methane-oxidizing bacterium H-2. Appl Environ Microbiol 52: 1403–1406PubMedGoogle Scholar
  43. Jones JW, Nagle DP, Whitman WR (1987) Methanogens and the diversity of archaebac-teria. Microbiol Rev 51: 135–177PubMedGoogle Scholar
  44. Kannen A, McCaffrey I, Bowman RL (1962) A flow-through method of scintillation counting of carbon-14 and tritium in gas-liquid Chromatographie effluents. J Lipid Res 3: 372–377Google Scholar
  45. Kenten RH (1956) Gasometric analysis in plant investigation (Warburg, van Slyke, micro-diffusion methods and ethylene). In:Paech K, Tracey MV (eds) Modern methods of plant analysis, vol 1. Springer, Berlin Göttingen Heidelberg, pp 415–451Google Scholar
  46. Keppler JG, Dijkstron G, Schols JA (1957) In:Destry DF (ed) Vapour phase chromatog-raphy, proceedings of the first symposium. Academic Press, New York London, p 222Google Scholar
  47. Krzycki JA, Zeikus JG (1984) Acetate catabolism by Methanosarcina barkeri: hydrogen-dependent methane production from acetate by a soluble cell protein fraction. FEMS Microbiol Lett 25: 27–32CrossRefGoogle Scholar
  48. Kyryacos G, Boord CE (1957) Separation of hydrogen, oxygen, nitrogen, methane, and carbon monoxide by gas adsorption chromatography. Anal Chem 29: 787–788CrossRefGoogle Scholar
  49. Laurinavichus KS, Belyaev SS (1979) Radioisotope method for determining microbial methane production rate. Mikrobiologiya (Engl Transi Plenum, New York) 47: 1115–1116Google Scholar
  50. Leak DJ, Dalton H (1983) In vivo studies of primary alcohols, aldehydes and carboxylic acids as electron donors for the methane mono-oxygenase in a variety of methano-trophs. J Gen Microbiol 129: 3487–3497Google Scholar
  51. Littlewood AB (1970) Gas chromatography, principle, techniques, and applications. Academic Press, New York London, pp 546Google Scholar
  52. Lovely DR, White RH, Ferry JG (1984) Identification of methyl coenzyme M as an intermediate in methanogenesis from acetate in Methanosarcina spp. J Bacteriol 160: 521–525Google Scholar
  53. Mah RA, Smith MR (1981) The methanogenic bacteria. In:Starr MP, Stolp H, Truper HG, Balows A, Schlegel HG (eds) The prokaryotes and handbook on habitats, isolation, and identification of bacteria. Springer, Berlin Heidelberg New York, pp 948–977Google Scholar
  54. Mah RA, Smith MR, Baresi L (1978) Studies on an acetate-fermenting strain of Methanosarcina. Appl Environ Microbiol 35: 1174–1184PubMedGoogle Scholar
  55. Martin RO (1968) Gas chromatograph-combustion-continuous counting system for analysis of microgram amounts of radioactive metabolites. Anal Biochem 40: 1197–1200Google Scholar
  56. McBride BC, Wolfe RS (1971) A new coenzyme of methyl transfer, coenzyme M. Biochemistry 10: 2317–2324PubMedCrossRefGoogle Scholar
  57. McPheat WL, Mann NH, Dalton H (1987) Isolation of mutants of the obligate methano-troph Methylomonas albus defective in growth on methane. Arch Microbiol 148: 40–43CrossRefGoogle Scholar
  58. Meijden P van der, Heythuysen HJ, Sliepenbeek HT, Houwen FP, Drift C van der, Vogels GD (1983) Activation and inactivation of methanol:2-mercaptoethanesulfonic acid methyltransferase from Methanosarcina barkeri. J Bacteriol 153: 6–11PubMedGoogle Scholar
  59. Meyers AJ (1980) Evalutation of bromoethane as a suitable analogue in methane oxidation studies. FEMS Microbiol Lett 9: 297–300CrossRefGoogle Scholar
  60. Naumann E, Fahlbusch K, Gottschalk G (1984) Presence of a trimethylamine:HS-coen-zyme M methyltransferase in Methanosarcina barkeri. Arch Microbiol 138: 79–83CrossRefGoogle Scholar
  61. Nelson DR, Zeikus JG (1974) Rapid method for the radioisotopic analysis of gaseous end products of anaerobic metabolism. Appl Microbiol 28: 258–261PubMedGoogle Scholar
  62. Nelson MK, Ferry JG (1984) Carbon monoxide-dependent methyl coenzyme M methyl-reductase in acetotrophic bacteria. J Bacteriol 160: 526–532PubMedGoogle Scholar
  63. Niosh Manual of analytical methods (1974) HEW Publ (NIOSH) 75-121 2nd edn, vol 1-7. Superintendent of documents, US Gov Print Off, Washington, DC (GPO No 1733-0041)Google Scholar
  64. Ohi K, Nishimura T, Okazaki M, Miura Y (1979) Determination of dissolved hydrogen concentration during cultivation and assimilation of gaseous substrates by Alcaligenes hydrogenophilus. J Ferment Technol 57: 203–209Google Scholar
  65. Ottenstein DM, Bartley DA (1971 a) Separation of free acids C2–C5 in dilute aqueous solution column technology. J Chromatogr Sci 9: 673–681Google Scholar
  66. Ottenstein DM, Bartley DA (1971b) Improved gas chromatography separation of free acids C2–C5 in dilute solution. Anal Chem 43: 952–955CrossRefGoogle Scholar
  67. Patel RN, Hou CT, Laskin AI, Felix A (1982) Microbial oxidation of hydrocarbons:properties of a soluble methane monooxygenase from a facultative methane-utilizing organism, Methylobacterium sp. strain CRL-26. Appl Environ Microbiol 44: 1130–1137PubMedGoogle Scholar
  68. Pine M, Barker HA (1956) Studies on the methane fermentation XII. The pathway of hydrogen in the acetate fermentation. J Bacteriol 71: 644–648Google Scholar
  69. Popjak G, Lowe AE, Moore D (1962) Scintillation counter for simultaneous assay of H3 and C14 in gas-liquid Chromatographic vapors. J Lipid Res 3: 364–371Google Scholar
  70. Purnell H (1962) Gas chromatography. Wiley, New York, pp 441Google Scholar
  71. Reeburgh WS (1980) Anaerobic methane oxidation; rate depth distribution in Skan Bay sediments. Earth Planet Sci Lett 47: 345–352CrossRefGoogle Scholar
  72. Ribbons DW (1975) Oxidation of C1 compounds by particulate fractions from Methyl-ococcus capsulatus: distribution and properties of methane-dependent reduced nicotin-amide adenine dinucleotide oxidase (methane hydroxylase). J Bacteriol 122: 1351–1363PubMedGoogle Scholar
  73. Ribbons DW, Michalover JL (1970) Methane oxidation by cell-free extracts of Methyl-ococcus capsulatus. FEBS Lett 11: 41–44PubMedCrossRefGoogle Scholar
  74. Robbins JD, Bakke JE (1967) Method for collecting 14CO2 from a hydrogen flame detector. J Gas Chromatogr 5: 525–526Google Scholar
  75. Robinson JA, Strayer RF, Tiedge JM (1981) Method for measuring dissolved hydrogen in anaerobic ecosystems:application to the rumen. Appl Environ Microbiol 41: 545548PubMedGoogle Scholar
  76. Rogosa M, Love LL (1968) Direct quantitative gas Chromatographic separation of C2–C6 fatty acids, methanol, and ethyl alcohol in aqueous microbial fermentation media. Appl Microbiol 16: 285–290PubMedGoogle Scholar
  77. Roitsch T, Stolp H (1985) Distribution of dissimilatory enzymes in methane and methanol oxidizing bacteria. Arch Microbiol 143: 233–236CrossRefGoogle Scholar
  78. Romesser JA, Balch WE (1980) Coenzyme M:preparation and assay. Meth Enzymol 67: 545–556PubMedCrossRefGoogle Scholar
  79. Salanitro JP, Muirhead PA (1975) Quantitative method for the gas Chromatographic analysis of short-chain monocarboxylic and dicarboxylic acids in fermentation media. Appl Microbiol 29: 374–381PubMedGoogle Scholar
  80. Simmons JH (1972) The use of gas chromatography to measure organic solvent in factory atmospheres. In:Perry SG (ed) Gas chromatography. Applied Science, England pp 17–24Google Scholar
  81. Smith MR, Lequerica JL (1985) Methanosarcina mutant unable to produce methane or assimilate carbon from acetate. J Bacteriol 164: 618–625PubMedGoogle Scholar
  82. Smith MR, Mah RA (1980) Acetate as sole carbon and energy source for growth of Methanosarcina strain 227. Appl Environ Microbiol 39: 992–999Google Scholar
  83. Stanley SH, Prior SH, Leak DJ, Dalton H (1983) Copper stress underlies the fundamental change in intracellular location of methane mono-oxygenase in methane-oxidizing organisms:studies in batch and continuous cultures. Biotech Lett 5: 487–92CrossRefGoogle Scholar
  84. Taylor GT, Pirt SJ (1977) Nutrition and factors limiting the growth of a methanogenic bacterium (Methanobacterium thermoautotrophicum). Arch Microbiol 113: 17–22PubMedCrossRefGoogle Scholar
  85. Tonge GM, Harrison DEF, Knowles CJ, Higgins IJ (1975) Properties and partial purification of the methane-oxidising enzyme system from Methylosinus trichosporium. FEBS Lett 58: 293–299PubMedCrossRefGoogle Scholar
  86. Umbreit WW, Burris RW, Stauffer JF (1964) Manometric techniques a manual describing methods applicable to the study of tissue metabolism. Burgess, Minneapolis, pp 5–24Google Scholar
  87. Walther R, Fahlbusch K, Sievert R, Gottschalk G (1981) Formation of trideuteromethane from deuterated trimethylamine or methylamine by Methanosarcina barkeri. J Bac-teriol 148: 371–373Google Scholar
  88. Washburn EW, West CJ, Dorsey NE, Bichowsky FR, Klemene A (eds) (1928) International critical tables of numeric data physics, chemistry and technology, vol 3. McGraw-Hill, New York, pp 254–269Google Scholar
  89. Watson JT (1972) Mass spectrometry instrumentation. In:Waller GR (ed) Biochemical applications of mass spectrometry. Wiley, New York London Sydney Toronto, pp 23–49Google Scholar
  90. Whittenbury R, Dalton H (1981) The methylotrophic bacteria. In:Starr MP, Stolp H, Truper HG, Balows A, Schlegel HG (eds) The prokaryotes a handbook on habitats, isolation and identification of bacteria. Springer, Berlin Heidelberg New York, pp 894–902Google Scholar
  91. Whittenbury R, Phillips KC, Wilkinson JF (1970) Enrichment, isolation and some properties of methane-utilizing bacteria. J Gen Microbiol 61: 205–218PubMedGoogle Scholar
  92. Whittenbury R, Dalton H, Eccleston M, Reed HL (1974) The different types of methane oxidizing bacteria and some of their more unusual properties. Proc Int Symp Microbiol growth on C1-compounds. Soc Ferment Technol, Osaka, Jpn, pp 1–9Google Scholar
  93. Wilhelm E, Battino R, Wilcock RJ (1977) Low pressure solubility of gases in liquid water. Chem Rev 77: 219–262CrossRefGoogle Scholar
  94. Wolfe RS, Higgins IJ (1979) Microbial biochemistry of methane — a study in contrasts. Int Rev Biochem 21: 267–353Google Scholar
  95. Zehnder AJB, Brock TD (1979) Methane formation and methane oxidation by methano-genic bacteria. J Bacteriol 137: 420–432PubMedGoogle Scholar
  96. Zehnder AJB, Wuhrmann K (1977) Physiology of a Methanobacterium strain AZ. Arch Microbiol 111: 199–205CrossRefGoogle Scholar
  97. Zehnder AJB, Huser B, Brock TD (1979) Measuring radioactive methane with the liquid scintillation counter. Appl Environ Microbiol 37: 897–899PubMedGoogle Scholar
  98. Zinder SH, Mah RA (1979) Isolation and characterization of a thermophilic strain of Methanosarcina unable to use H2-CO2 for methanogenesis. Appl Environ Microbiol 38: 996–1008PubMedGoogle Scholar
  99. Zinder SH, Cardwell SC, Anguish T, Lee M, Koch K (1984) Methanogenesis in a thermophilic (58°) anaerobic digestor:Methanothrix sp. as an important aceticlastic metha-nogen. Appl Environ Microbiol 47: 796–807PubMedGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 1989

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  • M. R. Smith
  • L. Baresi

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