1887

Abstract

168 was assayed for its growth on tricarboxylic acid (TCA) cycle intermediates and related compounds as the sole carbon sources. Growth of the organism was supported by citrate, D-isocitrate, succinate, fumarate and L-malate, whereas no growth was observed in the presence of -aconitate,2-oxoglutarate, D-malate, oxaloacetate and tricarballylate. Growth of the organism on the tricarboxylates citrate and D-isocitrate required the presence of functional CitM, an Mg–citrate transporter, whereas its growth on succinate, fumarate and L-malate appeared to be CitM-independent. Interestingly, the naturally occurring enantiomer D-isocitrate was favoured over L-isocitrate by the organism. Like citrate, D-isocitrate was shown to be an inducer of expression in . The addition of 1 mM Mg to the growth medium improved growth of the organism on both citrate and D-isocitrate, suggesting that D-isocitrate was taken up by CitM in complex with divalent metal ions. Subsequently, the ability of CitM to transport D-isocitrate was demonstrated by competition experiments and by heterologous exchange in right-side-out membrane vesicles prepared from cells expressing . None of the other TCA cycle intermediates and related compounds tested were recognized by CitM. Uptake experiments using radioactive Ni provided direct evidence that D-isocitrate is transported in complex with divalent metal ions.

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2002-11-01
2020-04-07
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References

  1. Asai K, Baik S. H, Kasahara Y, Moriya S., Ogasawara N. 2000; Regulation of the transport system for C4-dicarboxylic acids in Bacillus subtilis . Microbiology146:263–271
    [Google Scholar]
  2. Aymerich S, Gonzy-Tréboul G., Steinmetz M. 1986; 5′-Noncoding region sacR is the target of all identified regulation affecting the levansucrase gene in Bacillus subtilis . J Bacteriol166:993–998
    [Google Scholar]
  3. Bandell M., Lolkema J. S. 1999; Stereoselectivity of the membrane potential-generating citrate and malate transporters of lactic acid bacteria. Biochemistry38:10352–10360
    [Google Scholar]
  4. Bandell M, Ansanay V, Rachidi N, Dequin S., Lolkema J. S. 1997; Membrane potential-generating malate (MleP) and citrate (CitP) transporters of lactic acid bacteria are homologous proteins. Substrate specificity of the 2-hydroxycarboxylate transporter family. J Biol Chem272:18140–18146
    [Google Scholar]
  5. Boorsma A, van der Rest M. E, Lolkema J. S., Konings W. N. 1996; Secondary transporters for citrate and the Mg2+–citrate complex in Bacillus subtilis are homologous proteins. J Bacteriol178:6216–6222
    [Google Scholar]
  6. Bott M. 1997; Anaerobic citrate metabolism and its regulation in enterobacteria. Arch Microbiol167:78–88
    [Google Scholar]
  7. Fortnagel P., Freese E. 1968; Analysis of sporulation mutants. II. Mutants blocked in the citric acid cycle. J Bacteriol95:1431–1438
    [Google Scholar]
  8. Fournier R. E, McKillen M. N, Pardee A. B., Willecke K. 1972; Transport of dicarboxylic acids in Bacillus subtilis . Inducible uptake of l-malate. J Biol Chem247:5587–5595
    [Google Scholar]
  9. Goel A, Lee J, Domach M. M., Ataai M. M. 1995; Suppressed acid formation by cofeeding of glucose and citrate in Bacillus cultures: emergence of pyruvate kinase as a potential metabolic engineering site. Biotechnol Prog11:380–385
    [Google Scholar]
  10. Janausch I. G, Zientz E, Tran Q. H, Kröger A., Unden G. 2002; C4-dicarboxylate carriers and sensors in bacteria. Biochim Biophys Acta1553:39–56
    [Google Scholar]
  11. Kaback H. R. 1983; The lac carrier protein in Escherichia coli . J Membr Biol76:95–112
    [Google Scholar]
  12. Kay W. W. 1978; Transport of carboxylic acids. In Bacterial Transport pp385–411 Edited by Rosen B. P.. New York: Marcel Dekker;
    [Google Scholar]
  13. Krom B. P, Warner J. B, Konings W. N., Lolkema J. S. 2000; Complementary metal ion specificity of the metal–citrate transporters CitM and CitH of Bacillus subtilis . J Bacteriol182:6374–6381
    [Google Scholar]
  14. Krom B. P, Aardema R., Lolkema J. S. 2001; Bacillus subtilis YxkJ is a secondary transporter of the 2-hydroxycarboxylate transporter family that transports l-malate and citrate. J Bacteriol183:5862–5869
    [Google Scholar]
  15. Kunst F, Ogasawara N, Moszer I.. 48 other authors 1997; The complete genome sequence of the Gram-positive bacterium Bacillus subtilis . Nature390:249–256
    [Google Scholar]
  16. Lolkema J. S, Enequist H., van der Rest M. E. 1994; Transport of citrate catalyzed by the sodium-dependent citrate carrier of Klebsiella pneumoniae is obligatorily coupled to the transport of two sodium ions. Eur J Biochem220:469–475
    [Google Scholar]
  17. Martell A. E., Smith R. M. 1977; Critical Stability Constants, vol. 3: Other Organic Ligands New York: Plenum;
    [Google Scholar]
  18. McKillen M. N, Willecke K., Pardee A. B. 1972; Citrate transport by Bacillus subtilis . In The Molecular Basis of Biological Transport pp249–70 Edited by Woessner J. F., Huijung F.. New York: Academic Press;
    [Google Scholar]
  19. Miller J. H. 1972; Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Somers J. M, Sweet G. D., Kay W. W. 1981; Fluorocitrate resistant tricarboxylate transport mutants of Salmonella typhimurium . Mol Gen Genet181:338–345
    [Google Scholar]
  21. Sweet G. D, Kay C. M., Kay W. W. 1984; Tricarboxylate-binding proteins of Salmonella typhimurium . Purification, crystallization, and physical properties. J Biol Chem259:1586–1592
    [Google Scholar]
  22. van der Rest M. E, Molenaar D., Konings W. N. 1992; Mechanism of Na+-dependent citrate transport in Klebsiella pneumoniae . J Bacteriol174:4893–4898
    [Google Scholar]
  23. Warner J. B., Lolkema J. S. 2002; LacZ-promoter fusions: the effect of growth. Microbiology148:1241–1243
    [Google Scholar]
  24. Warner J. B, Krom B. P, Magni C, Konings W. N., Lolkema J. S. 2000; Catabolite repression and induction of the Mg2+–citrate transporter CitM of Bacillus subtilis . J Bacteriol182:6099–6105
    [Google Scholar]
  25. Wei Y, Guffanti A. A, Ito M., Krulwich T. A. 2000; Bacillus subtilis YqkI is a novel malic/Na+–lactate antiporter that enhances growth on malate at low protonmotive force. J Biol Chem275:30287–30292
    [Google Scholar]
  26. Widenhorn K. A, Somers J. M., Kay W. W. 1988; Expression of the divergent tricarboxylate transport operon ( tctI) of Salmonella typhimurium . J Bacteriol170:3223–3227
    [Google Scholar]
  27. Yamamoto H, Murata M., Sekiguchi J. 2000; The CitST two-component system regulates the expression of the Mg–citrate transporter in Bacillus subtilis . Mol Microbiol37:898–912
    [Google Scholar]
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