Influence of sulphur-containing compounds on the growth of Methanosarcina barkeri in a defined medium

  • Paul Scherer
  • Hermann Sahm
Industrial Microbiology


Optimal growth of Methanosarcina barkeri occurred in a defined medium containing methanol when 2.5–4 mM sodium sulphide was added giving a concentration of 0.04–0.06 mM dissolved sulphide (HS+S2−. When the sulphide concentration was too low for optimal growth (e.g., 0.1 mM Na2S added) the addition of the redox resin ‘Serdoxit’ acted as a sulphide reservoir and caused a significant stimulation of growth. Furthermore it could be demonstrated that iron sulphide, zinc sulphide or L-methionine could also act as sulphur sources while the addition of sodium sulphate to sulphide-depleted media failed to restore growth. The amino acid L-cysteine (0.85 mM) stimulated growth but could not replace Na2S.

Under optimal cysteine-and sulphide concentrations the generation time of this strain was about 7–9 h during growth on methanol, giving a growth yield of about 0.14 g/g methanol consumed. Different M. barkeri strains were also able to grow under these conditions on acetate (30–50 h doubling time) without a significant lag-phase and with complete substrate consumption even though the inoculum was grown on methanol or H2−CO2. When methanol and acetate were present as a mixture in the medium both were used simultaneously.


Sulphide Sodium Sulphate Doubling Time Optimal Growth Growth Yield 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Balch WE, Wolfe RS (1979) Specificity and biological distribution of coenzyme M (2-mercaptoethanesulfonic acid). J Bacteriol 137:256–263Google Scholar
  2. Barker HA (1940) Studies upon the methane fermentation. IV. The isolation and culture of Methanobacterium omelianskii. Antonie van Leeuwenhoek J Microbiol Serol 6:201–220Google Scholar
  3. Bock R, Puff H-J (1968) Bestimmung von Sulfid mit einer Sulfidionenempfindlichen Elektrode. Z Anal Chem 240:381–386Google Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Google Scholar
  5. Breden CR, Buswell AM (1933) The use of shredded asbestos in methane fermentations. J Bacteriol 26:379–384Google Scholar
  6. Bryant MP, Tzeng SF, Robinson IM, Joyner AE Jr (1971) Nutrient requirements of methanogenic bacteria. In: Pohland FG (ed) Advances in chemistry series 105. Am Chem Soc, Washington DC, pp 23–40Google Scholar
  7. Cappenberg TE (1975) A study of mixed continuous cultures of sulfate-reducing and methane-producing bacteria. Microbial Ecol 2:60–72Google Scholar
  8. Chiapelli F, Vasil A, Haggerty DF (1979) The protein concentration of crude cell and tissue extracts as estimated by the method of dye binding: Comparison with the Lowry method. Anal Biochem 94:160–165Google Scholar
  9. D'Ans, Lax E (1967) Taschenbuch für Chemiker und Physiker, vol I. Springer, Berlin Heidelberg New York, pp 872–873Google Scholar
  10. Eirich LD, Vogels GD, Wolfe RS (1979) Distribution of coenzyme F420 and properties of its hydrolytic fragments. J Bacteriol 140:20–27Google Scholar
  11. Hansson G (1979) Effects of carbon dioxide and methane on methanogenesis. Eur J Appl Microbiol Biotechnol 6:351–359Google Scholar
  12. Hippe H, Caspari D, Fiebig K, Gottschalk G (1979) Utilization of trimethylamine and other N-methyl compounds for growth and methane formation by Methanosarcina barkeri. Proc Natl Acad Sci USA 76:494–498Google Scholar
  13. Khan AW (1980) Degradation of cellulose to methane by a coculture of Acetivibrio cellulolyticus and Methanosarcina barkeri. FEMS Microbiol Lett 9:233–235Google Scholar
  14. Koch OG, Koch-Dedic GA (1974) Handbuch der Spurenanalyse, vol II. Springer, Berlin Heidelberg New York, pp 1004–1012Google Scholar
  15. Mah RA, Ward DM, Baresi L, Glass TL (1977) Biogenesis of methane. Annu Rev Microbiol 31:309–341Google Scholar
  16. Mah RA, Smith MR, Baresi L (1978) Studies on an acetatefermenting strain of Methanosarcina. Appl Environ Microbiol 35:1174–1184Google Scholar
  17. Mah RA, Smith MR, Ferguson T, Zinder S (1980) Methanogenesis from H2−CO2, methanol and acetate by Methanosarcina. In: Proc 3rd Int Sympos on Microbial Growth on C1 Compounds. Sheffield (in press)Google Scholar
  18. McBride BC, Wolfe RS (1971) A new coenzyme of methyltransfer, coenzyme M. Biochemistry 10:2317–2324Google Scholar
  19. Miller TL, Wolin MJ (1974) A serum bottle modification of the Hungate technique for cultivating obligate anaerobes. Appl Microbiol 27:985–987Google Scholar
  20. Miller TL, Wolin MJ (1980) Molybdate and sulfide inhibit H2 and increase formate production from glucose by Ruminococcus albus. Arch Microbiol 124:137–142Google Scholar
  21. Mountfort DO, Asher RA (1979) Effect of inorganic sulfide on the growth and metabolism on Methanosarcina barkeri strain DM. Appl Environ Microbiol 37:670–675Google Scholar
  22. Oppermann RA, Nelson WO, Brown RE (1957) In vitro studies on methanogenic rumen bacteria. J Dairy Sci 40:779–788Google Scholar
  23. Scherer P, Sahm H (1979) Züchtung von Methanosarcina barkeri auf Methanol oder Acetat in einem definierten Medium. In: Dellweg H (ed) Proc 4. Sympos Techn Mikrobiologie Berlin. Schmacht-Difodruck, Bamberg, pp 281–290Google Scholar
  24. Scherer P, Sahm H (1980) Growth of Methanosarcina barkeri on methanol or acetate in a defined medium. In: Stafford DA, Wheatley BI (eds) Proc 1 st Int Sympos on Anaerobic Digestion, University College Cardiff 1979. Scientific Press, Cardiff, pp 45–47Google Scholar
  25. Schnellen CGTP (1947) Onderzoekingen over de methaangisting. Thesis, Technical University DelftGoogle Scholar
  26. Smith MR, Mah RA (1978) Growth and methanogenesis by Methanosarcina strain 227 on acetate and methanol. Appl Environ Microbiol 36:870–879Google Scholar
  27. Smith MR, Mah RA (1980) Acetate as sole carbon and energy source for growth of Methanosarcina strain 227. Appl Environ Microbiol 39:993–999Google Scholar
  28. Smith MR, Zinder SH, Mah RA (1980) Microbial methanogenesis from acetate. Process Biochem 15:34–39Google Scholar
  29. Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41: 100–180Google Scholar
  30. Weimer PJ, Zeikus JG (1978) Acetate metabolism in Methanosarcina barkeri. Arch Microbiol 119:175–182Google Scholar
  31. Wellinger A, Wuhrmann K (1977) Influence of sulfide compounds on the metabolism of Methanobacterium strain AZ. Arch Microbiol 115:13–17Google Scholar
  32. Winter JW, Wolfe RS (1979) Complete degradation of carbohydrate to carbon dioxide and methane by syntrophic cultures of Acetobacterium woodii and Methanosarcina barkeri. Arch Microbiol 121:97–102Google Scholar
  33. Wolin EA, Wolin MJ, Wolfe RS (1963) Formation of methane by bacterial extracts. J Biol Chem 238:2882–2886Google Scholar
  34. Zehnder AJB, Wuhrmann K (1976) Titanium(III) citrate as a nontoxic oxidation-reduction buffering system for the culture fo obligate anaerobes. Science 190:1165–1166Google Scholar
  35. Zehnder AJB (1978) Ecology of methane formation. In: Mitchell CR (ed) Water pollution microbiology, vol II. Wiley J, New York, pp 349–376Google Scholar
  36. Zeikus JG (1977) The biology of methanogenic bacteria. Bacteriol Rev 41:514–541Google Scholar
  37. Zhilina TN, Zavarzin GA (1973) Trophic relationships between Methanosarcina and its associates. (Engl Transl) Mikrobiologiya 2:266–273Google Scholar
  38. Zhilina TN (1978) Growth of a pure Methanosarcina culture, biotype 2, on acetate. (Engl Transl) Mikrobiologiya 47:321–323Google Scholar
  39. Zinder SH, Mah RA (1979) Isolation and characterization of a thermophilic strain of Methanosarcina unable to use H2−CO2 for methanogenesis. Appl Environ Microbiol 38:996–1008Google Scholar

Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • Paul Scherer
    • 1
  • Hermann Sahm
    • 1
  1. 1.Institut für Biotechnologie der Kernforschungsanlage JülichJülichFederal Republic of Germany

Personalised recommendations