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Clostridium neopropionicum sp. nov., a strict anaerobic bacterium fermenting ethanol to propionate through acrylate pathway

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Abstract

Strain X4 was isolated several years ago from an anaerobic mesophilic plant treating vegetable cannery waste waters. It was the first example of propionic fermentation from ethanol. Morphologic and physiologic characterizations of the strain are presented here. This strain is described as type strain of a new species, Clostridium neopropionicum sp. nov. Whole cells of strain X4 ferment [1-13C]ethanol and CO2 to [2-13C]propionate, [1-13C]acetate and [2-13C]propanol, suggesting the absence of a randomizing pathway during the propionate formation. Enzymes involved in this fermentation were assayed in cell-free extracts of cells grown with ethanol as sole substrate. Alcohol dehydrogenase, aldehyde dehydrogenase, phosphate acetyl transferase, acetate kinase, pyruvate synthase, lactate dehydrogenases, and the enzymes of the acrylate pathway were detected at activities sufficient to be involved in ethanol fermentation. The same pathway may be used for the degradation of lactate or acrylate to acetate.

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References

  • Bauchop T, Elsden (1960) The growth of microorganisms in relation to their energy supply. J Gen Microbiol 23: 457–469

    Google Scholar 

  • Bergsma J, vanDongen MBM, Konings WN (1982) Purification and characterization of NADH dehydrogenase from Bacillus subtilis. Eur J Biochem 128: 151–157

    Google Scholar 

  • Bradford NM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Google Scholar 

  • Bridger WA, Ramaley RF, Boyer PD (1969) Succinyl coenzyme A synthetase from Escherichia coli. Methods Enzymol 13: 70–75

    Google Scholar 

  • Bryant MP, Wolin EA, Wolin MJ, Wolfe RS (1967) Methanobacillus omelianskii, a symbiotic association of two species of bacteria. Arch Microbiol 59: 20–31

    Google Scholar 

  • Bryant MP, Campbell LL, Reddy CA, Crabil MR (1977) Growth of Desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria. Appl Environ Microbiol 33: 1162–1169

    Google Scholar 

  • Buckel W, Dorn U, Semmler R (1981) Gluconate CoA-transferase from Acidaminococcus fermentans. Eur J Biochem 118: 315–321

    Google Scholar 

  • Cardon BP, Barker HA (1946) Two new amino-acid-fermenting bacteria, Clostridium propionicum and Diplococcus glycinophilus. J Bacteriol 52: 629–634

    Google Scholar 

  • Cato EP, George WL, Finegold SM (1986) Genus Clostridium. In: Sneath PHA, Mair NS, Sharpe ME, Holt JG (eds) Bergeys's manual of systematic bacteriology, vol II. Williams and Wilkins, Baltimore, pp 1141–1200

    Google Scholar 

  • Decker K, Jungermann K, Thauer RK (1970) Energy production in anaerobic organisms. Angew Chem Int Ed Eng 9: 138–158

    Google Scholar 

  • Doddema HJ, Vogels GD (1978) Improved identification of methanogenic bacteria by fluorescence microscopy. Appl Environ Microbiol 36: 752–754

    Google Scholar 

  • Dörner C, Schink B (1990) Clostridium homopropionicum sp. nov., a new strict anaerobe growing with 2-, 3-, or 4-hydroxybutyrate. Arch Microbiol 154: 342–348

    Google Scholar 

  • Dubourguier HC, Samain E, Prensier G, Albagnac G (1986) Characterization of two strains of Pelobacter carbinolicus isolated from anaerobic digesters. Arch Microbiol 145: 248–253

    Google Scholar 

  • Gottschalk G (1979) Bacterial metabolism. Springer Berlin Heidelberg New York

    Google Scholar 

  • Hobbe JE, Daley RJ, Jasper S (1977) Use of nucleopore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol 33: 1225–1228

    Google Scholar 

  • Johnson JL (1981) Genetic characterization. In: Gerhardt P, Murray RGE, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (eds) Manual of methods for general bacteriology. American Society for Microbiology, Washington DC, pp 450–472

    Google Scholar 

  • König H, Buckel W, Langworthy TA (1985) Ultrastructure of the cell envelope and amino acid composition of the murein of Clostridium symbiosum. FEMS Microbiol Lett 30: 283–288

    Google Scholar 

  • Kuchta RD, Abeles RH (1985) Lactate reduction in Clostridium propionicum. J Biol Chem 260: 13181–13189

    Google Scholar 

  • Laanbroek HJ, Abee T, Voogd IL (1982) Alcohol conversion by Desulfobulbus propionicus Lindhorst in the presence and absence of sulfate and hydrogen. Arch Microbiol 133: 178–184

    Google Scholar 

  • Mahler HR (1955) Butyryl coenzyme A dehydrogenase. Methods Enzymol 1: 553–559

    Google Scholar 

  • Melville SB, Michel TA, Macy JM (1988) Pathway and sites for energy conservation in the metabolism of glucose by Selenomonas ruminantium. J Bacteriol 170: 5298–5304

    Google Scholar 

  • Miller TL, Wolin MJ (1974) A serum bottle modification of the Hungate technique for cultivating obligate anaerobes. Appl Environ Microbiol 27: 985–987

    Google Scholar 

  • Nakajima H, Suzuki K, Imahori K (1978) Purification and properties of acetate kinase from Bacillus stearothermophilus. J Biochem 84: 193–203

    Google Scholar 

  • Oberlies G, Fuchs G, Thauer RK (1980) Acetate thiokinase and the assimilation of acetate in Methanobacterium thermoautotrophicum. Arch Microbiol 128: 248–252

    Google Scholar 

  • Pankhania IP, Spormann AM, Hamilton WA, Thauer RK (1988) Lactate conversion to acetate, CO2, and H2 in cell suspensions of Desulfovibrio vulgaris (Marburg): indications for the involvement of an energy driven reaction. Arch Microbiol 150: 26–31

    Google Scholar 

  • Peck HDJr, Gest H (1956) A new procedure for assay of bacterial hydrogenases. J Bacteriol 71: 70–80

    Google Scholar 

  • Phelps TJ, Zeikus JG (1984) Influence of pH on terminal carbon metabolism in anoxic sediments from a midly acidic lake. Appl Environ Microbiol 48: 1088–1095

    Google Scholar 

  • Samain E, Albagnac D, Dubourguier HC, Touzel JP (1982) Characterization of a new propionic acid bacterium that ferments ethanol and displays a growth factor-dependant association with a gram-negative homoacetogen. FEMS Microbiol Lett 15: 69–74

    Google Scholar 

  • Schink B (1984) Fermentation of 2,3-butanediol by Pelobacter carbinolicus sp. nov. and Pelobacter propionicus sp. nov., and evidence for propionate formation from C2 compounds. Arch Microbiol 137: 33–41

    Google Scholar 

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

    Google Scholar 

  • Schink B, Phelps TJ, Eichler B, Zeikus ZG (1985) Comparison of ethanol degradation pathways in anoxic freshwater environments. J Gen Microbiol 131: 651–660

    Google Scholar 

  • Schink B, Schlegel HG (1979) The membrane-bound hydrogenase of Alcaligenes eutrophus. I. Solubilization, purification, and biochemical properties. Biochim Biophys Acta 567: 315–324

    Google Scholar 

  • Schütz H, Radler F (1984) Propanediol-1,2-dehydratase and metabolism of glycerol of Lactobacillus brevis. Arch Microbiol 139: 366–370

    Google Scholar 

  • Schweiger G, Buckel W (1984) On the dehydration of (R)-lactate in the fermentation of alanine to propionate by Clostridium propionicum. FEBS Lett 171: 79–84

    Google Scholar 

  • Schweiger G, Buckel W (1985) Identification of acrylate, the product of the dehydration of (R)-lactate catalysed by cell-free extracts from Clostridium propionicum. FEBS Lett 185: 253–256

    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 

  • Stouthamer AH, Bettenhaussen C (1973) Utilization of energy for growth and maintenance in continuous and batch cultures of microorganisms. Biochim Biophys Acta 301: 53–70

    Google Scholar 

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

    Google Scholar 

  • Tholozan JL, Samain E, Grivet JP, Moletta R, Dubourguier HC, Albagnac G (1988a) Reductive carboxylation of propionate to butyrate in methanogenic ecosystems. Appl Environ Microbiol 54: 441–445

    Google Scholar 

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

    Google Scholar 

  • Tholozan JL, Samain E, Grivet JP, Albagnac G (1990) Propionate metabolism in a methanogenic enrichment culture. Direct reductive carboxylation and acetogenesis pathways. FEMS Microbiol Ecol 73: 291–298

    Google Scholar 

  • Toraya T, Honda S, Fukui S (1979) Fermentation of 1,2-propanediol and 1,2-ethanediol by some genera of Enterobacteriaceae, involving coenzyme B12-dependent diol dehydratase. J Bacteriol 139: 39–47

    Google Scholar 

  • Touzel JP, Petroff D, Albagnac G (1985) Isolation and characterization of a new thermophilic Methanosarcina, the strain CHTI 55. Syst Appl Microbiol 6: 66–71

    Google Scholar 

  • Zeikus JG, Fuchs G, Kenealy W, Thauer RK (1977) Oxidoreductases involved in cell carbon synthesis of Methanobacterium thermoautotrophicum. J Bacteriol 132: 604–613

    Google Scholar 

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Authors and Affiliations

  1. Station de Technologie Alimentaire, Institut National de la Recherche Agronomique, 369, rue J. Guesde, BP 39, F-59 651, Villeneuve d'Ascq CEDEX, France

    J. L. Tholozan, J. P. Touzel & G. Albagnac

  2. C.E.R.M.A.V., Centre National de la Recherche Scientifique, BP 53X, F-38 041, Grenoble CEDEX, France

    E. Samain

  3. Centre de Biophysique Moléculaire, Centre National de la Recherche Scientifique, 1 A, avenue de la Recherche Scientifique, F-45 071, Orléans CEDEX 2, France

    J. P. Grivet

  4. Laboratoire de Microbiologie, les Cézeaux, Université Blaise Pascal, F-63177, Aubière CEDEX, France

    G. Prensier

Authors
  1. J. L. Tholozan
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  2. J. P. Touzel
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  3. E. Samain
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  4. J. P. Grivet
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  5. G. Prensier
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  6. G. Albagnac
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Tholozan, J.L., Touzel, J.P., Samain, E. et al. Clostridium neopropionicum sp. nov., a strict anaerobic bacterium fermenting ethanol to propionate through acrylate pathway. Arch. Microbiol. 157, 249–257 (1992). https://doi.org/10.1007/BF00245158

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  • DOI: https://doi.org/10.1007/BF00245158

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