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. 1976 Feb 1;153(2):173–179. doi: 10.1042/bj1530173

Factors affecting the activity of citrate synthase of Acetobacter xylinum and its possible regulatory role.

M Swissa, M Benziman
PMCID: PMC1172560  PMID: 6002

Abstract

The citrate synthase activity of Acetobacter xylinum cells grown on glucose was the same as of cells grown on intermediates of the tricarboxylic acid cycle. The activity of citrate synthase in extracts is compatible with the overall rate of acetate oxidation in vivo. The enzyme was purified 47-fold from sonic extracts and its molecular weight was determined to be 280000 by gel filtration. It has an optimum activity at pH 8.4. Reaction rates with the purified enzyme were hyperbolic functions of both acetyl-CoA and oxaloacetate. The Km for acetyl-CoA is 18 mum and that for oxaloacetate 8.7 mum. The enzyme is inhibited by ATP according to classical kinetic patterns. This inhibition is competitive with respect to acetyl-CoA (Ki = 0.9 mM) and non-competitive with respect to oxaloacetate. It is not affected by changes in pH and ionic strength and is not relieved by an excess of Mg2+ ions. Unlike other Gram-negative bacteria, the A. xylinum enzyme is not inhibited by NADH, but is inhibited by high concentrations of NADPH. The activity of the enzyme varies with energy charge in a manner consistent with its role in energy metabolism. It is suggested that the flux through the tricarboxylic acid cycle in A. xylinum is regulated by modulation of citrate synthase activity in response to the energy state of the cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkinson D. E. The energy charge of the adenylate pool as a regulatory parameter. Interaction with feedback modifiers. Biochemistry. 1968 Nov;7(11):4030–4034. doi: 10.1021/bi00851a033. [DOI] [PubMed] [Google Scholar]
  2. BENZIMAN M., ABELIOVITZ A. METABOLISM OF DICARBOXYLIC ACIDS IN ACETOBACTER XYLINUM. J Bacteriol. 1964 Feb;87:270–277. doi: 10.1128/jb.87.2.270-277.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BENZIMAN M., BURGER-RACHAMIMOV H. Synthesis of cellulose from pyruvate by succinate-grown cells of Acetobacter xylinum. J Bacteriol. 1962 Oct;84:625–630. doi: 10.1128/jb.84.4.625-630.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benziman M., Eizen N. Pyruvate-phosphate dikinase and the control of gluconeogenesis in Acetobacter xylinum. J Biol Chem. 1971 Jan 10;246(1):57–61. [PubMed] [Google Scholar]
  5. Benziman M. Factors afecting the activity of pyruvate kinase of Acetobacter xylinum. Biochem J. 1969 May;112(5):631–636. doi: 10.1042/bj1120631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benziman M., Levy L. Phosphorylation coupled to malate oxidation in Acetobacter xylinum. Biochem Biophys Res Commun. 1966 Jul 20;24(2):214–217. doi: 10.1016/0006-291x(66)90722-4. [DOI] [PubMed] [Google Scholar]
  7. Benziman M. Role of phosphoenolpyruvate carboxylation in Acetobacter xylinum. J Bacteriol. 1969 Jun;98(3):1005–1010. doi: 10.1128/jb.98.3.1005-1010.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Flechtner V. R., Hanson R. S. Coarse and fine control of citrate synthase from Bacillus subtilis. Biochim Biophys Acta. 1969 Jul 30;184(2):252–262. doi: 10.1016/0304-4165(69)90027-0. [DOI] [PubMed] [Google Scholar]
  9. Flechtner V. R., Hanson R. S. Regulation of the tricarboxylic acid cycle in bacteria. A comparison of citrate synthases from different bacteria. Biochim Biophys Acta. 1970 Nov 24;222(2):253–264. doi: 10.1016/0304-4165(70)90114-5. [DOI] [PubMed] [Google Scholar]
  10. GROMET Z., SCHRAMM M., HESTRIN S. Synthesis of cellulose by Acetobacter Xylinum. 4. Enzyme systems present in a crude extract of glucose-grown cells. Biochem J. 1957 Dec;67(4):679–689. doi: 10.1042/bj0670679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Garland P. B. Control of citrate synthesis in mitochondria. Biochem Soc Symp. 1968;27:41–60. [PubMed] [Google Scholar]
  12. Jangaard N. O., Unkeless J., Atkinson D. E. The inhibition of citrate synthase by adenosine triphosphate. Biochim Biophys Acta. 1968 Jan 8;151(1):225–235. doi: 10.1016/0005-2744(68)90177-0. [DOI] [PubMed] [Google Scholar]
  13. Johnson D. E., Hanson R. S. Bacterial citrate synthases: purification, molecular weight and kinetic mechanism. Biochim Biophys Acta. 1974 Jun 18;350(2):336–353. doi: 10.1016/0005-2744(74)90508-7. [DOI] [PubMed] [Google Scholar]
  14. Kosicki G. W., Lee L. P. Effect of divalent metal ions on nucleotide inhibition of pig heart citrate synthase. J Biol Chem. 1966 Aug 10;241(15):3571–3574. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. MONOD J., CHANGEUX J. P., JACOB F. Allosteric proteins and cellular control systems. J Mol Biol. 1963 Apr;6:306–329. doi: 10.1016/s0022-2836(63)80091-1. [DOI] [PubMed] [Google Scholar]
  17. Moellering H., Gruber W. Determination of citrate with citrate lyase. Anal Biochem. 1966 Dec;17(3):369–376. doi: 10.1016/0003-2697(66)90172-2. [DOI] [PubMed] [Google Scholar]
  18. SCHRAMM M., HESTRIN S. Factors affecting production of cellulose at the air/liquid interface of a culture of Acetobacter xylinum. J Gen Microbiol. 1954 Aug;11(1):123–129. doi: 10.1099/00221287-11-1-123. [DOI] [PubMed] [Google Scholar]
  19. Srere P. A. An eclectic view of metabolic regulation: control of citrate synthase activity. Adv Enzyme Regul. 1970;9:221–233. doi: 10.1016/s0065-2571(71)80046-8. [DOI] [PubMed] [Google Scholar]
  20. Srere P. A. Studies on purified citrate-enzymes: metabolic interpretations. Biochem Soc Symp. 1968;27:11–21. [PubMed] [Google Scholar]
  21. Weinhouse H., Benziman M. Regulation of gluconeogenesis in Acetobacter xylinum. Eur J Biochem. 1972 Jun 23;28(1):83–88. doi: 10.1111/j.1432-1033.1972.tb01886.x. [DOI] [PubMed] [Google Scholar]
  22. Weinhouse H., Benziman M. Regulation of hexose phosphate metabolism in Acetobacter xylinum. Biochem J. 1974 Mar;138(3):537–542. doi: 10.1042/bj1380537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weitzman P. D., Dunmore P. Citrate synthases: allosteric regulation and molecular size. Biochim Biophys Acta. 1969 Jan 7;171(1):198–200. doi: 10.1016/0005-2744(69)90122-3. [DOI] [PubMed] [Google Scholar]
  24. Weitzman P. D.J., Dunmore P. Regulation of citrate synthase activity by alpha-ketoglutarate. Metabolic and taxonomic significance. FEBS Lett. 1969 Jun;3(4):265–267. doi: 10.1016/0014-5793(69)80154-7. [DOI] [PubMed] [Google Scholar]
  25. Weitzman P. D., Jones D. Regulation of citrate synthase and microbial taxonomy. Nature. 1968 Jul 20;219(5151):270–272. doi: 10.1038/219270a0. [DOI] [PubMed] [Google Scholar]

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