Skip to main content
SpringerLink
Log in

Anaerobic degradation of isobutyrate by methanogenic enrichment cultures and by a Desulfococcus multivorans strain

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

Abstract

Methanogenic enrichment cultures with isobutyrate as sole source of carbon and energy were inoculated with sediment and sludge samples from freshwater and marine origin. Over more than 20 transfers, these cultures fermented 2 mol isobutyrate with 1 mol CO2 via an intermediate formation of n-butyrate to 4 mol acetate and 1 mol CH4. The primary isobutyrate-fermenting bacteria could not be purified. From one of the marine enrichment cultures, a sulfate-reducing bacterium was isolated which oxidized isobutyrate with sulfate completely to CO2. Based on its physiological and morphological properties, this strain was assigned to the known species Desulfococcus multivorans. It also oxidized many other fatty acids without significant release of short-chain intermedeates. The enzymes involved in isobutyrate degradation by this bacterium were assayed in cell-free extracts. The results indicate that isobutyrate is activated to its CoA derivative and oxidized via methylmalonate semialdehyde to propionyl-CoA. Propionyl-CoA is further converted via the methylmalonyl-CoA pathway to acetyl-CoA which is finally cleaved by the CO-dehydrogenase system. It is evident that this is not the pathway used by the fermenting bacteria prevailing in the methanogenic enrichment cultures. There results are discussed on the basis of energetical considerations.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allison MJ (1969) Biosynthesis of amino acids by ruminal microorganisms. J Anim Sci 29:797–807

    Google Scholar 

  • Allison MJ (1978) Production of branched-chain volatile fatty acids by certain anaerobic bacteria. Appl Environ Microbiol 35: 872–877

    Google Scholar 

  • Allison MJ, Bryant MP (1963) Biosynthesis of branched-chain amino acids from branched-chain fatty acids by rumen bacteria. Arch Biochem Biophys 101:262–277

    Google Scholar 

  • Barker HA (1981) Amino acid degradation by anaerobic bacteria. Ann Rev Biochem 50:23–40

    Google Scholar 

  • Bergmeyer HU (ed) (1974) Methoden der enzymatischen Analyse, 3rd edn. Verlag Chemie, Weinheim

    Google Scholar 

  • Bernt E, Bergmeyer HU (1974) Isocitrate dehydrogenase. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, vol 1. Verlag Chemie, Weinheim, pp 664–667

    Google Scholar 

  • Brandis-Heep A, Gebhardt NA, Thauer RK, Widdel F, Pfennig N (1983) Anaerobic acetate oxidation to CO2 by Desulfobacter postgatei. 1. Demonstration of all enzymes required for the operation of the citric acid cycle. Arch Microbiol 136:222–229

    Google Scholar 

  • Cline JD (1969) Spectrophotometric determination of hydrogen sulfide in natural waters. Limnol Oceanogr 14:454–458

    Google Scholar 

  • Green DE, Mii S, Mahler HR, Bock RM (1954) Studies on the fatty acid oxidizing system of animal tissue. III. Butyryl coenzyme A dehydrogenase. J Biol Chem 206:1–12

    Google Scholar 

  • Gregersen T (1978) Rapid method for distinction of Gram-negative from Gram-positive bacteria. Eur J Appl Microbiol Biotechnol 5:123–127

    Google Scholar 

  • Gustafson WG, Feinberg BA, McFarland JT (1986) Energetics of β-oxidation. Reduction potentials of general fatty acyl-CoA dehydrogenase, electron transfer flavoprotein, and fatty acyl-CoA substrates. J Biol Chem 261:7733–7741

    Google Scholar 

  • Harwood CS, Canale-Parola E (1981) Branched-chain amino acid fermentation by a marine spirochete: strategy for starvation survival. J Bacteriol 148:109–116

    Google Scholar 

  • Hilpert W, Schink B, Dimroth P (1984) Life by a new decarboxylation-dependent energy conservation mechanism with Na+ as coupling ion. EMBO J 3:1665–1670

    Google Scholar 

  • Knappe J, Blaschkowski HP, Gröbner P, Schmit T (1974) Pyruvate formate lyase of Escherichia coli: the acetyl-enzyme intermediate. Eur J Biochem 50:253–263

    Google Scholar 

  • Koch M, Dolfing J, Wuhrmann K, Zehnder AJB (1983) Pathways of propionate degradation by enriched methanogenic cultures. Appl Environ Microbiol 45:1411–1414

    Google Scholar 

  • Kuenen JG, Veldkamp H (1972) Thiomicrospira pelophila gen. nov. sp. n., a new obligately chemolithotrophic colourless sulfur bacterium. Antonie van Leeuwenhoek J Microbiol Serol 38:241–256

    Google Scholar 

  • Lee MJ, Zinder SH (1988) Isolation and characterization of a thermophilic bacterium which oxidizes acetate in syntrophic association with a methanogen and which grows acetogenically on H2−CO2. Appl Environ Microbiol 54:124–129

    Google Scholar 

  • Lovley DR, Klug MJ (1982) Intermediary metabolism of organic matter in the sediments of a eutrophic lake. Appl Environ Microbiol 43:552–560

    Google Scholar 

  • Magee CM, Rodeheaver G, Edgerton MT, Edlich RF (1975) A more reliable Gram staining technic for diagnosis of surgical infections. Am J Surgery 130:341–346

    Google Scholar 

  • Massey LK, Sokatch JR, Conrad RS (1976) Branched-chain amino acid catabolism in bacteria. Bacteriol Rev 40:42–54

    Google Scholar 

  • Moskowitz GJ, Merrick JM (1969) Metabolism of poly-β-hydroxybutyrate. II. Enzymatic synthesis of d-(-)β-hydroxybutyryl coenzyme A by an enoyl hydrase from Rhodospirillum rubrum. Biochemistry 8:2748–2754

    Google Scholar 

  • Pfennig N (1978) Rhodocyclus purpureus gen. nov. and sp. nov., a ring-shaped, vitamin B12-requiring member of the family Rhodospirillaceae. Int J Syst Bacteriol 28:283–288

    Google Scholar 

  • Rendina G, Coon MJ (1957) Enzymatic hydrolysis of the coenzyme A thiol esters of β-hydroxypropionic and β-hydroxyisobutyric acids. J Biol Chem 225:523–534

    Google Scholar 

  • Robinson WG, Coon MJ (1957) The purification and properties of β-hydroxyisobutyric dehydrogenase. J Biol Chem 225:511–521

    Google Scholar 

  • Robinson WG, Nagle R, Bachhawat BK, Kupiecki FP, Coon MJ (1957) Coenzyme A thiol esters of isobutyric, methacrylic, and β-hydroxyisobutyric acids as intermediates in the enzymatic degradation of valine. J Biol Chem 224:1–11

    Google Scholar 

  • Sansone FJ, Martens CS (1981) Determination of volatile fatty acid turnover rates in organic-rich marine sediments. Mar Chem 10:233–247

    Google Scholar 

  • Sansone FJ, Martens CS (1982) Volatile fatty acid cycling in organicrich marine sediments. Geochim Cosmochim Acta 46:1575–1589

    Google Scholar 

  • Schauder R, Eikmanns B, Thauer RK, Widdel F, Fuchs G (1986) Acetate oxidation to CO2 in anaerobic bacteria via a novel pathway not involving reactions of the citric acid cycle. Arch Microbiol 145:162–172

    Google Scholar 

  • Schink B (1985a) Fermentation of acetylene by an obligate anaerobe, Pelobacter acetylenicus sp. nov. Arch Microbiol 142:295–301

    Google Scholar 

  • Schink B (1985b) Mechanism and kinetics of succinate and propionate degradation in anoxic freshwater sediments and sewage sludge. J Gen Microbiol 131:643–650

    Google Scholar 

  • Schink B, Pfennig N (1982) Fermentation of trihydroxybenzenes by Pelobacter acidigallici gen. nov. sp. nov., a new strictly anaerobic, non-sporeforming bacterium. Arch Microbiol 133:195–201

    Google Scholar 

  • Schink B, Thauer RK (1988) Energetics of syntrophic methane formation and the influence of aggregation. In: Lettinga G, Zehnder AJB, Grotenhuis JTC, Hulshoff Pol LW (eds) Granular anaerobic sludge; microbiology and technology. Pudoc, Wageningen, pp 5–17

    Google Scholar 

  • Schink B, Kremer DR, Hansen ThA (1987) Pathway of proprionate formation from ethanol in Pelobacter propionicus. Arch Microbiol 147:321–327

    Google Scholar 

  • Scrutton MC, Olmsted MR, Utter MF (1969) Pyruvate carboxylase from chicken liver. Meth Enzymol 13:235–249

    Google Scholar 

  • Shelton DR, Tiedje JM (1984) Isolation and partial characterization of bacteria in an anaerobic consortium that mineralizes 3-chlorobenzoic acid. Appl Environ Microbiol 48:840–848

    Google Scholar 

  • Sokatch JR (1966) Alanine and aspartate formation during grwoth on valine-14C by Pseudomonas aeruginosa. J Bacteriol 92:72–81

    Google Scholar 

  • Sokatch JR, Sanders LE, Marshall VP (1968) Oxidation of methylmalonate semialdehyde to propionyl coenzyme A in Pseudomonas aeruginosa grown on valine. J Biol Chem 243:2500–2506

    Google Scholar 

  • Stams AJM, Kremer DR, Nicolay K, Weenk GH, Hansen TA (1984) Pathway of propionate formation in Desulfobulbus propionicus. Arch Microbiol 139:167–173

    Google Scholar 

  • Stieb M, Schink B (1984) A new 3-hydroxybutyrate-fermenting anaerobe, Ilyobacter polytropus gen. nov. sp. nov., possessing various fermentation pathways. Arch Microbiol 140:139–146

    Google Scholar 

  • Stieb M, Schink B (1985) Anaerobic oxidation of fatty acids by Clostridium bryantii so. nov., a sporeforming, obligately syntrophic bacterium. Arch Microbiol 140:387–390

    Google Scholar 

  • Stieb M, Schink B (1986) Anaerobic degradation of isovalerate by a defined methanogenic coculture. Arch Microbiol 144:291–295

    Google Scholar 

  • Tanner RS, Wolfe RS (1988) Nutritional requirements of Methanomicrobium mobile. Appl Environ Microbiol 54:625–628

    Google Scholar 

  • Thauer RK, Morris JG (1984) Metabolism of chemotrophic anaerobes: old views and new aspects. In: Kelly DP, Carr NG (eds) The microbe 1984 Part II. Prokaryotes and eukaryotes. Soc Gen Microbiol Symp, vol 36. Cambridge University Press, Cambridge, pp 123–168

    Google Scholar 

  • Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180

    Google Scholar 

  • Tholozan J-L, Samain E, Grivet J-P (1988) Isomerization between n-butyrate and isobutyrate in enrichment cultures. FEMS Microbiol Ecol 53:187–191

    Google Scholar 

  • Widdel F (1988) Microbiology and ecology of sulfate- and sulfurreducing bacteria. In: Zehnder AJB (ed) Environmental biology of anaerobes. John Wiley, New York (in press)

    Google Scholar 

  • Widdel F, Pfennig N (1981) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments. Description of Desulfobacter postgatei gen. nov. sp. nov. Arch Microbiol 129:395–400

    Google Scholar 

  • Widdel F, Pfennig N (1984) Dissimilatory sulfate- or sulfuroxidizing bacteria. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 1. Williams and Wilkins, Baltimore London, pp 663–679

    Google Scholar 

  • Widdel F, Kohring G-W, Mayer F (1983) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. III. Characterization of the filamentous gliding Desulfonema limicola gen. nov. sp. nov., and Desulfonema magnum sp. nov. Arch Microbiol 134:286–294

    Google Scholar 

  • Wofford NQ, Beaty PS, McInerney MJ (1986) Preparation of cellfree extracts and the enzymes involved in fatty acid metabolism in Syntrophomonas wolfei. J Bacteriol 167:179–185

    Google Scholar 

  • Zinder SH, Koch M (1984) Non-aceticlastic methanogenesis from acetate: acetate oxidation by a thermophilic syntrophic coculture. Arch Microbiol 138:263–272

    Google Scholar 

  • Zinder SH, Cardwell SC, Anguish T, Lee M, Koch M (1984) Methanogenesis in a thermophilic (58°C) anaerobic digestor: Methanothrix sp. as an important aceticlastic methanogen. Appl Environ Microbiol 47:796–807

    Google Scholar 

Download references

Author information

Author notes

  1. Marion Stieb

    Present address: Space Development Division, Dornier System GmbH, D-7990, Friedrichshafen 1, FRG

  2. Bernhard Schink

    Present address: Lehrstuhl Mikrobiologie I, Auf der Morgenstelle 28, D-7400, Tübingen, FRG

Authors and Affiliations

  1. Fakultät für Biologie, Universität Konstanz, Postfach 5560, D-7750, Konstanz, Federal Republic of Germany

    Marion Stieb & Bernhard Schink

Authors
  1. Marion Stieb
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bernhard Schink
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stieb, M., Schink, B. Anaerobic degradation of isobutyrate by methanogenic enrichment cultures and by a Desulfococcus multivorans strain. Arch. Microbiol. 151, 126–132 (1989). https://doi.org/10.1007/BF00414426

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00414426

Key words

Search

Navigation