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The CO2 assimilation via the reductive tricarboxylic acid cycle in an obligately autotrophic, aerobic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus

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Abstract

The incorporation of 14CO2 by the cell suspensions of an extremely thermophilic, aerobic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus was studied. After short time incubation of the cell suspensions with 14CO2, the radiactivity was initially present in aspartate, glutamate, succinate, phosphorylated compounds, citrate, malate and fumarate. All of these compounds except phosphorylated compounds were related to the members of the tricarboxylic acid cycle. The proportion of labelled aspartate onglutamate in total radioactivity on each chromatogram decreased with incubation time, while the percentage of the radioactivity incorporated in phosphorylated compounds increased with time up to 10 s. These indicated that aspartate and glutamate is derived from primary products of CO2 fixation.

In cell-free extracts of Hydrogenobacter thermophilus, the two key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase and phosphoribulokinase could not be detected. The key enzymes of the reductive tricarboxylic acid cycle, fumarate reductase and ATP citrate lyase were present. Activities of phosphoenolpyruvate synthetase and pyruvate carboxylase were also detected. The referse reactions (dehydrogenase reactions) of α-ketoglutarate synthase and pyruvate synthase could be detected by using methyl viologen as an electron acceptor.

These findings strongly suggested that a new type of the reductive tricarboxylic acid cycle operated as the CO2 fixation pathway in Hydrogenobacter thermophilus.

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References

  • Antranikian G, Herzberg C, Gottschalk G (1982) Characterization of ATP citrate lyase from Chlorobium limicola. J Bacteriol 152:1284–1287

    PubMed  Google Scholar 

  • Bassham JA, Calvin M (1957) The path of carbon in photosynthesis. Prentice-Hall Inc, Englewood Cliffs, NJ

    Google Scholar 

  • Bowien B, Schlegel HG (1981) Physiology and biochemistry of acrobic hydrogen-oxidizing bacteria. Ann Rev Microbiol 35:405–452

    Article  Google Scholar 

  • Buchanan BB, Evans MCW (1965) The synthesis of α-ketoglutarate from succinate and carbon dioxide by a subcellular preparation of a photosynthetic bacterium. Proc Natl Acad Sci USA 54:1212–1218

    PubMed  Google Scholar 

  • Cooper RA, Kornberg HL (1974) Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase. In: Boyer PD (ed) The enzymes, 3rd edn, vol 10. Academic Press, New York, pp 631–649

    Google Scholar 

  • Daniels L, Zeikus JG (1978) One-carbon metabolism in methanogenic bacteria: Analysis of short-term fixation products of 14CO2 and 14CH3OH incorporated into whole cells. J Bacteriol 136:75–84

    PubMed  Google Scholar 

  • Eden G, Fuchs G (1982) Total synthesis of acetyl coenzyme A involved in autotrophic CO2 fixation in Acetobacterium woodii. Arch Microbiol 133:66–74

    Google Scholar 

  • Eden G, Fuchs G (1983) Autotrophic CO2 fixation in Acetobacterium woodii. II. Demonstration of enzymes involved. Arch Microbiol 135:68–73

    Google Scholar 

  • Evans MCV, Buchanan BB (1965) Photoreduction of ferredoxin and its use in carbon dioxide fixation by a subcellular system from a photosynthetic bacterium. Proc Natl Acad Sci USA 53:1420–1425

    PubMed  Google Scholar 

  • Evans MCW, Buchanan BB (Arnon DI (1966) A new ferredoxindependent carbon reduction cycle in a photosynthetic bacterium. Proc Natl Acad Sci USA 55:928–934

    PubMed  Google Scholar 

  • Eyzaguirre J, Jansen K, Fuchs G (1982) Phosphoenolpyruvate synthetase in Methanobacterium thermoautotrophicum. Arch Microbiol 132:67–74

    Google Scholar 

  • Fuchs G, Stupperich E (1978) Evidence for an incomplete reductive carboxylic acid cycle in Methanobacterium thermoautotrophicum. Arch Microbiol 118:121–125

    PubMed  Google Scholar 

  • Fuchs G, Stupperich E (1980) Acetyl CoA, a central intermediate of autotrophic CO2 fixation in Methanobacterium thermoautotrophicum. Arch Microbiol 127:267–272

    Google Scholar 

  • Fuchs G, Stupperich E (1982) Autotrophic CO2 fixation pathway in Methanobacterium thermoautotrophicum. Zbl Bakt Hyg, 1 Abt Orig C 3:277–288

    Google Scholar 

  • Fuchs G, Stupperich E (1983) CO2 fixation pathway in bacteria. Physiol Veg 21:845–854

    Google Scholar 

  • Fuchs G, Stupperich E, Thauer RK (1978) Acetate assimilation and the synthesis of alanine, aspartate and glutamate in Methanobacterium thermoautotrophicum. Arch Microbiol 117:61–66

    Google Scholar 

  • Fuchs G, Stupperich E, Jaenchen R (1980a) Autotrophic CO2 fixation in Chlorobium limicola. Evidence against the operation of the Calvin cycle in growing cells. Arch Microbiol 128:56–63

    Google Scholar 

  • Fuchs G, Stupperich E, Eden G (1980b) Autotrophic CO2 fixation in Chlorobium limicola. Evidence for the operation of a reductive tricarboxylic acid cycle in growing cells. Arch Microbiol 128:64–71

    Google Scholar 

  • Fuchs G, Winter H, Steiner I, Stupperich E (1983) Enzymes of gluconeogenesis in the autotroph Methanobacterium thermoautotrophicum. Arch Microbiol 136:160–162

    Google Scholar 

  • Ishii M, Kawasumi T, Igarashi Y, Kodama T, Minoda Y (1983) 2-Methylthio-1,4-naphthoquinone, a new quinone from an extremely thermophilic hydrogen bacterium. Agric Biol Chem 47:167–169

    Google Scholar 

  • Ivanovsky RN, Sintsov NV, Kondratieva EN (1980) ATP-linked citrate lyase activity in the green sulfur bacterium Chorobium limicola forma thiosulfatophilum. Arch Microbiol 128:239–241

    Google Scholar 

  • Jansen K, Stupperich E, Fuchs G (1982) Carbohydrate synthesis from acetyl CoA in the autotroph Methanobacterium thermoautotrophicum. Arch Microbiol 132:355–364

    Google Scholar 

  • Kandler O, Stetter KO (1981) Evidence for autotrophic CO2 assimilation in Sulfolobus brierleyi via a reductive carboxylic acid pathway. Zbl Bakt Hyg, I Abt Orig C 2:111–121

    Google Scholar 

  • Kawasumi T, Igarashi Y, Kodama T, Minoda Y (1980) Isolation of strictly thermophilic and obligately autotrophic hydrogen bacteria. Agric Biol Chem 44:1985–1986

    Google Scholar 

  • Kawasumi T, Igarashi Y, Kodama T, Minoda Y (1984) Hydrogenobacter thermophilus gen. nov., sp. nov., an extremely thermophilic, aerobic, hydrogen-oxidizing bacterium. Int J Syst Bacteriol 34:5–10

    Google Scholar 

  • Lowry OH, Roeebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  Google Scholar 

  • Myers WF, Huang KY (1969) Thin-layer chromatography of citric acid cycle compounds. In: Lowenstein JM (ed) Methods in enzymology, vol 13. Academic Press, New York, pp 431–434

    Google Scholar 

  • Shiba H, Kawasumi T, Igarashi Y, Kodama T, Minoda Y (1982) The deficient carbohydrate metabolic pathways and the incomplete tricarboxylic acid cycle in an obligately autotrophic hydrogen-oxidizing bacterium. Agric Biol Chem 46:2341–2345

    Google Scholar 

  • Shiba H, Kawasumi T, Igarashi Y, Kodama T, Minoda Y (1984) Effect of organic compounds on the growth of an obligately autotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6. Agric Biol Chem 48:2809–2813

    Google Scholar 

  • Siebert K, Schobert P, Bowien B (1981) Purification, some catalytic and molecular properties of phosphoribulokinase from Alcaligenes cutrophus. Biochim Biophys Acta 658:35–44

    PubMed  Google Scholar 

  • Stupperich E, Fuchs G (1981) Products of CO2 fixation and 14C labelling pattern of alanine in Methanobacterium thermoautotrophicum pulse-labelled with 14CO2. Arch Microbiol 136:294–300

    Google Scholar 

  • Stupperich E, Fuchs G (1983) Autotrophic acetyl coenzyme A synthesis in vitro from two CO2 in Methanobacterium. FEBS Lett 156:345–348

    Article  Google Scholar 

  • Stupperich E, Hammel KE, Fuchs G, Thauer RK (1983) Carbon monoxide fixation into the carboxyl group of acetyl coenzyme A during autotrophic growth of Methanobacterium. FEBS Lett 152:21–23

    Article  PubMed  Google Scholar 

  • Takeda Y, Suzuki F, Inoue H (1969) ATP citrate lyase (citrate-cleavage enzyme). In: Lowenstein JM (ed) Methods in enzymology, vol. 13. Academic Press, New York, pp 153–160

    Google Scholar 

  • Zeikus JG (1983) Metabolism of one-carbon compounds by chemotrophic anaerobes. Adv Microbial Physiol 24:215–299

    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

    PubMed  Google Scholar 

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Shiba, H., Kawasumi, T., Igarashi, Y. et al. The CO2 assimilation via the reductive tricarboxylic acid cycle in an obligately autotrophic, aerobic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus . Arch. Microbiol. 141, 198–203 (1985). https://doi.org/10.1007/BF00408058

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  • DOI: https://doi.org/10.1007/BF00408058

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