Abstract
Chlorofluexus aurantiacus OK-70 fl was grown photoautotrophically with hydrogen as the electron source. The lowest doubling time observed was 26 h.
The mechanism of CO2 fixation in autotrophically grown cells was studied. The presence of ribulose-1,5-bis-phosphate carboxylase and phosphoribulokinase could not be demonstrated. Carbon isotope fractionation (δ13C) was small, and alanine and aspartate but not 3-phosphoglycerate were the major labelled compounds in short term 14CO2 labelling. Thus CO2 is not fixed by the Calvin cycle.
Fluoroacetate (FAc) completely inhibited protein synthesis in cultures and caused a slight citrate accumulation. However, CO2 fixation continued and increased polyglucose formation occurred. Under these conditions added acetate was metabolized to polyglucose, as were glycine, serine, glyoxylate and succinate, but to a lesser extent; little or no formate or CO was utilised.
Glyoxylate inhibited CO2 fixation in vivo, indicating that pyruvate is formed from acetyl-CoA and CO2 by pyruvate synthase. Two key enzymes of the reductive TCA cycle, citrate lyase and α-ketoglutarate synthase were not detected in cell free extracts, but pyruvate synthase and phosphoenolpyruvate carboxylase were demonstrated. It is concluded that acetyl-CoA is a central intermediate in the CO2 fixation process, but the mechanism of its synthesis is not clear.
Similar content being viewed by others
Abbreviations
- Rubisco:
-
ribulose-1,5-bisphosphate carboxylase
- TCA cycle:
-
tricarboxylic acid cycle
- FAc:
-
monofluoroacetate
- PEP:
-
phosphoenolpyruvate
- MV:
-
methyl viologen
- TTC:
-
triphenyltetrazolium chloride
- PMS:
-
phenazine methosulfate
References
Allen TH, Root WS (1955) Colorimetric determination of carbon monoxide in air by an improved palladium chloride method. J Biol Chem 216:309–317
Berg P (1956) Acyladenylates: An enzymatic mechanism of acetate activation. J Biol Chem 222:991–1013
Buchanan BB (1972) Ferredoxin linked carboxylation reactions. In: Boyer PD (ed) The enzymes, 3. edn, vol VI. Academic Press, New York, pp 193–216
Cooper RA, Kornberg HL (1974) Phosphoenolpyruvate synthetase and pyruvate phosphate dikinase. In: Boyer PD (ed) The enzymes, 3. edn, vol 10. Academic Press, New York, pp 631–649
Craigh HH (1957) Isotope standards for carbon and oxygen and correction factors for mass spectrometric analysis for carbon dioxide. Geochim Cosmochim Acta 12:133–149
Dieckert G, Hansch M, Conrad R (1984) Acetate synthesis from 2 CO2 in acetogenic bacteria: is carbon monoxide an intermediate? Arch Microbiol 138:224–228
Drutschmann M, Klemme JH (1985) Sulfide repressed, membrane bound hydrogenase in the thermophilic facultative phototroph, Chloroflexus aurantiacus. FEMS Microbiol Lett 28:231–235
Dürre P, Andreesen JR (1982) Pathway of carbon dioxide reduction to acetate without a net energy requirement in Clostridium purinolyticum. FEMS Microbiol Lett 15:51–56
Evans MCW, Buchanan BB, Arnon DI (1966) A new ferredoxin dependent carbon reduction cycle in a photosynthetic bacterium. Proc Natl Acad Sci USA 55:928–934
Fox GE, Stackebrandt E, Hespell RB, Gibson J, Maniloff J, Dyer TA, Wolfe RS, Balch WE, Tanner RS, Magrum LJ, Zablen LB, Blakemore R, Gupta R, Bonen L, Lewis BJ, Stahl DA, Luehrsen KR, Chen KN, Woese CR (1980) The phylogeny of procaryotes. Science 209:457–463
Fuchs G, Schnitker U, Thauer RK (1974) Carbon monoxide oxidation by growing cultures of Clostridium pasteurianum. Eur J Biochem 49:111–115
Fuchs G, Stupperich E, Eden G (1980) Autotrophic CO2 fixation in Chlorobium limicola. Evidence for the operation of a reductive tricarboxylic acid cycle in growing cells. Arch Microbiol 128:64–71
Hall NP, Cornelivs MJ, Keys AJ (1983) The enzymatic determination of bicarbonate and CO2 in reagents and buffer solutions. Anal Biochem 132:152–157
Hart BA, Gibson J (1975) Ribulose-5-phosphate kinase from Chromatium. In: Wood WA (ed) Methods in enzymology, vol XLII. Academic Press, New York, pp 115–119
Hatch MD, Slack RC (1970) Photosynthetic CO2 fixation pathways. Ann Rev Plant Physiol 21:141–162
Knowles FC, Benson AA (1983) The biochemistry of arsenic. Trends Biochem Sci 8:178–180
Lowry OH, Rosenbrough MS, Farr AL, Randall RS (1951) Protein measurements with the Folin phenol reagent. J Biol Chem 193:265–275
Løken Ø, Sirevåg R (1982) Evidence for the presence of the glyoxylate cycle in Chloroflexus. Arch Microbiol 132:276–279
Madigan M (1976) Studies on the physiological ecology of Chloroflexus aurantiacus, a filamentous photosynthetic bacterium. Ph. D. disseration, University of Wisconsin-Madison.
Madigan MT, Petersen SR, Brock TD (1984) Nutritional studies on Chloroflexus, a filamentous, photosynthetic, gliding bacterium. Arch Microbiol 100:97–103
Massey V (1955) Fumarase. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 1. Academic Press, New York, pp 729–735
Ochoa S (1955) Crystalline condensing enzyme from pig heart. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 1. Academic Press, New York, pp 685–694
Ormerod JG (1980) Photosynthesis of acetate from carbon dioxide. 3rd Int Symp on Microbial Growth on C1 Compounds, Sheffield, Abstracts of posters: 99–100
Park R, Epstein S (1960) Carbon isotope fractionation during photosynthesis. Geochim Cosmochim Acta 21:110–126
Peters RA, Wakelin RW, Rivett DEA, Thomas LC (1953) Fluoroacetae poisoning: Comparison of synthetic fluorocitric acid with the enzymically synthesized fluorotricarboxylic acid. Nature 171:1111–1112
Quayle JR (1972) The metabolism of one-carbon compounds in microorganisms. Adv Microb Physiol 7:119–203
Raeburn S, Rabinowitz JC (1971) Pyruvate: Ferredoxin oxido-reductase 1. The pyruvate-CO2 exchange reaction. Arch Biochem Biophys 146:9–20
Rose IA, O'Connell EL (1967) Mechanism of aconitase action. J Biol Chem 1870–1879
Schürmann P (1969) Separation of phosphate esters and algal extracts by thin-layer electrophoresis and chromatography. J Chromat 39:507–509
Scrutton MC, Young MR (1972) Pyruvate carboxylase. In: Boyer PD (ed) The enyzmes, 3. edn, vol 6. Academic Press, New York, pp 1–35
Siebert K, Schobert P, Bowien B (1981) Purification, some catalytic and molecular properties of phosphoribulokinase from Alcaligenes eutrophus. Biochim Biophys Acta 658:35–44
Sirevåg R (1974) Further studies on carbon dioxide fixation in green sulphur bacteria. Arch Microbiol 98:3–18
Sirevåg R (1975) Photoassimilation of acetate and metabolism of carbohydrate in Chlorobium thiosulfatophilum. Arch Microbiol 104:105–111
Sirevåg R, Castenholz R (1979) Aspects of carbon metabolism in Chloroflexus. Arch Microbiol 120:151–153
Sirevåg R, Ormerod JG (1970) Carbon dioxide fixation in green sulfur bacteria. Biochem J 120:399–408
Sirevåg R, Buchanan BB, Berry JA, Troughton JH (1977) Mechanisms of CO2 fixation in bacterial photosynthesis studied by the carbon isotope fractionation technique. Arch Microbiol 112:35–38
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
Tabita RF, McFadden B, Pfennig N (1974) d-Ribulose 1,5-biphosphate carboxylase in Chlorobium thiosulfatophilum Tassajara. Biochim Biophys Acta 341:187–194
Taylor TG (1953) A modified procedure for the microdetermination of citric acid. Biochem J 54:48–49
Thauer RK, Rupprecht E, Jungermann K (1970a) Glyoxylate inhibition of clostridial pyruvate synthase. FEBS Lett 9:271–273
Thauer RK, Rupprecht E, Jungermann K (1970b) The synthesis of one carbon units from CO2 via a new ferredoxin dependent monocarboxylic acid cycle. FEBS Lett 8:304–307
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Holo, H., Sirevåg, R. Autotrophic growth and CO2 fixation of Chloroflexus aurantiacus . Arch. Microbiol. 145, 173–180 (1986). https://doi.org/10.1007/BF00446776
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00446776