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. 1995 Jan;177(2):277–282. doi: 10.1128/jb.177.2.277-282.1995

Catabolic and anabolic enzyme activities and energetics of acetone metabolism of the sulfate-reducing bacterium Desulfococcus biacutus.

P H Janssen 1, B Schnik 1
PMCID: PMC176588  PMID: 7814315

Abstract

Acetone degradation by cell suspensions of Desulfococcus biacutus was CO2 dependent, indicating initiation by a carboxylation reaction, while degradation of 3-hydroxybutyrate was not CO2 dependent. Growth on 3-hydroxybutyrate resulted in acetate accumulation in the medium at a ratio of 1 mol of acetate per mol of substrate degraded. In acetone-grown cultures no coenzyme A (CoA) transferase or CoA ligase appeared to be involved in acetone metabolism, and no acetate accumulated in the medium, suggesting that the carboxylation of acetone and activation to acetoacetyl-CoA may occur without the formation of a free intermediate. Catabolism of 3-hydroxybutyrate occurred after activation by CoA transfer from acetyl-CoA, followed by oxidation to acetoacetyl-CoA. In both acetone-grown cells and 3-hydroxybutyrate-grown cells, acetoacetyl-CoA was thioyltically cleaved to two acetyl-CoA residues and further metabolized through the carbon monoxide dehydrogenase pathway. Comparison of the growth yields on acetone and 3-hydroxybutyrate suggested an additional energy requirement in the catabolism of acetone. This is postulated to be the carboxylation reaction (delta G(o)' for the carboxylation of acetone to acetoacetate, +17.1 kJ.mol-1). At the intracellular acyl-CoA concentrations measured, the net free energy change of acetone carboxylation and catabolism to two acetyl-CoA residues would be close to 0 kJ.mol of acetone-1, if one mol of ATP was invested. In the absence of an energy-utilizing step in this catabolic pathway, the predicted intracellular acetoacetyl-CoA concentration would be 10(13) times lower than that measured. Thus, acetone catabolism to two acetyl-CoA residues must be accompanied by the utilization of teh energetic equivalent of (at lease) one ATP molecule. Measurement of enzyme activities suggested that assimilation of acetyl-CoA occurred through a modified citric acid cycle in which isocitrate was cleaved to succinate and glyoxylate. Malate synthase, condensing glyoxylate and acetyl-CoA, acted as an anaplerotic enzyme. Carboxylation of pyruvate of phosphoenolpyruvate could not be detected.

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Selected References

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  1. Biegert T., Altenschmidt U., Eckerskorn C., Fuchs G. Enzymes of anaerobic metabolism of phenolic compounds. 4-Hydroxybenzoate-CoA ligase from a denitrifying Pseudomonas species. Eur J Biochem. 1993 Apr 1;213(1):555–561. doi: 10.1111/j.1432-1033.1993.tb17794.x. [DOI] [PubMed] [Google Scholar]
  2. Boynton Z. L., Bennett G. N., Rudolph F. B. Intracellular Concentrations of Coenzyme A and Its Derivatives from Clostridium acetobutylicum ATCC 824 and Their Roles in Enzyme Regulation. Appl Environ Microbiol. 1994 Jan;60(1):39–44. doi: 10.1128/aem.60.1.39-44.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Colby G. D., Chen J. S. Purification and properties of 3-hydroxybutyryl-coenzyme A dehydrogenase from Clostridium beijerinckii ("Clostridium butylicum") NRRL B593. Appl Environ Microbiol. 1992 Oct;58(10):3297–3302. doi: 10.1128/aem.58.10.3297-3302.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Diekert G. B., Thauer R. K. Carbon monoxide oxidation by Clostridium thermoaceticum and Clostridium formicoaceticum. J Bacteriol. 1978 Nov;136(2):597–606. doi: 10.1128/jb.136.2.597-606.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dörner C., Schink B. Clostridium homopropionicum sp. nov., a new strict anaerobe growing with 2-, 3-, or 4-hydroxybutyrate. Arch Microbiol. 1990;154(4):342–348. doi: 10.1007/BF00276529. [DOI] [PubMed] [Google Scholar]
  7. Friedrich M., Schink B. Hydrogen formation from glycolate driven by reversed electron transport in membrane vesicles of a syntrophic glycolate-oxidizing bacterium. Eur J Biochem. 1993 Oct 1;217(1):233–240. doi: 10.1111/j.1432-1033.1993.tb18238.x. [DOI] [PubMed] [Google Scholar]
  8. Fukui T., Ito M., Tomita K. Purification and characterization of acetoacetyl-CoA synthetase from Zoogloea ramigera I-16-M. Eur J Biochem. 1982 Oct;127(2):423–428. doi: 10.1111/j.1432-1033.1982.tb06889.x. [DOI] [PubMed] [Google Scholar]
  9. Gorny N., Schink B. Anaerobic degradation of catechol by Desulfobacterium sp. strain Cat2 proceeds via carboxylation to protocatechuate. Appl Environ Microbiol. 1994 Sep;60(9):3396–3400. doi: 10.1128/aem.60.9.3396-3400.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grupe H., Gottschalk G. Physiological Events in Clostridium acetobutylicum during the Shift from Acidogenesis to Solventogenesis in Continuous Culture and Presentation of a Model for Shift Induction. Appl Environ Microbiol. 1992 Dec;58(12):3896–3902. doi: 10.1128/aem.58.12.3896-3902.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kreke B., Cypionka H. Protonmotive force in freshwater sulfate-reducing bacteria, and its role in sulfate accumulation in Desulfobulbus propionicus. Arch Microbiol. 1992;158(3):183–187. doi: 10.1007/BF00290814. [DOI] [PubMed] [Google Scholar]
  12. Lack A., Fuchs G. Carboxylation of phenylphosphate by phenol carboxylase, an enzyme system of anaerobic phenol metabolism. J Bacteriol. 1992 Jun;174(11):3629–3636. doi: 10.1128/jb.174.11.3629-3636.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lewis A. J., Miller J. D. Keto acid metabolism in Desulfovibrio. J Gen Microbiol. 1975 Oct;90(2):286–292. doi: 10.1099/00221287-90-2-286. [DOI] [PubMed] [Google Scholar]
  14. Lewis A. J., Miller J. D. The tricarboxylic and acid pathway in Desulfovibrio. Can J Microbiol. 1977 Jul;23(7):916–921. doi: 10.1139/m77-135. [DOI] [PubMed] [Google Scholar]
  15. Oberlies G., Fuchs G., Thauer R. K. Acetate thiokinase and the assimilation of acetate in methanobacterium thermoautotrophicum. Arch Microbiol. 1980 Dec;128(2):248–252. doi: 10.1007/BF00406167. [DOI] [PubMed] [Google Scholar]
  16. Platen H., Schink B. Anaerobic degradation of acetone and higher ketones via carboxylation by newly isolated denitrifying bacteria. J Gen Microbiol. 1989 Apr;135(4):883–891. doi: 10.1099/00221287-135-4-883. [DOI] [PubMed] [Google Scholar]
  17. Platen H., Schink B. Enzymes involved in anaerobic degradation of acetone by a denitrifying bacterium. Biodegradation. 1990;1(4):243–251. doi: 10.1007/BF00119761. [DOI] [PubMed] [Google Scholar]
  18. Platen H., Schink B. Methanogenic degradation of acetone by an enrichment culture. Arch Microbiol. 1987;149(2):136–141. doi: 10.1007/BF00425079. [DOI] [PubMed] [Google Scholar]
  19. Platen H., Temmes A., Schink B. Anaerobic degradation of acetone by Desulfococcus biacutus spec. nov. Arch Microbiol. 1990;154(4):355–361. doi: 10.1007/BF00276531. [DOI] [PubMed] [Google Scholar]
  20. Rössle M., Kreusch J., Decker K. Acetyl coenzyme A and coenzyme a contents of growing clostridium kluyveri as determined by isotope assays. Arch Microbiol. 1981 Dec;130(4):288–293. doi: 10.1007/BF00425942. [DOI] [PubMed] [Google Scholar]
  21. TRUEPER H. G., SCHLEGEL H. G. SULPHUR METABOLISM IN THIORHODACEAE. I. QUANTITATIVE MEASUREMENTS ON GROWING CELLS OF CHROMATIUM OKENII. Antonie Van Leeuwenhoek. 1964;30:225–238. doi: 10.1007/BF02046728. [DOI] [PubMed] [Google Scholar]
  22. Thauer R. K. Citric-acid cycle, 50 years on. Modifications and an alternative pathway in anaerobic bacteria. Eur J Biochem. 1988 Oct 1;176(3):497–508. doi: 10.1111/j.1432-1033.1988.tb14307.x. [DOI] [PubMed] [Google Scholar]
  23. Thauer R. K., Jungermann K., Decker K. Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev. 1977 Mar;41(1):100–180. doi: 10.1128/br.41.1.100-180.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wofford N. Q., Beaty P. S., McInerney M. J. Preparation of cell-free extracts and the enzymes involved in fatty acid metabolism in Syntrophomonas wolfei. J Bacteriol. 1986 Jul;167(1):179–185. doi: 10.1128/jb.167.1.179-185.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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