1887

Microbiology

Volume 148, Issue 11

Growth of on citrate and isocitrate is supported by the Mg–citrate transporter CitM Free

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.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): divalent metal ion–citrate complex , exchange , FCCP, carbonylcyanide p-trifluormethoxy-phenylhydrazone , membrane vesicles , MSMYE, minimal salts medium/0·05% yeast extract , promoter fusion , RSO, right-side-out , TCA cycle intermediate and TCA, tricarboxylic acid
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-11-3405
2002-11-01
2019-10-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/11/1483405a.html?itemId=/content/journal/micro/10.1099/00221287-148-11-3405&mimeType=html&fmt=ahah

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. Microbiology 146, 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 Bacteriol 166, 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. Biochemistry 38, 10352-10360.[CrossRef]
    [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 Chem 272, 18140-18146.[CrossRef]
    [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 Bacteriol 178, 6216-6222.
     [Google Scholar]
  6. Bott, M. ( 1997; ). Anaerobic citrate metabolism and its regulation in enterobacteria. Arch Microbiol 167, 78-88.[CrossRef]
    [Google Scholar]
  7. Fortnagel, P. & Freese, E. ( 1968; ). Analysis of sporulation mutants. II. Mutants blocked in the citric acid cycle. J Bacteriol 95, 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 Chem 247, 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 Prog 11, 380-385.[CrossRef]
    [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 Acta 1553, 39-56.[CrossRef]
    [Google Scholar]
  11. Kaback, H. R. ( 1983; ). The lac carrier protein in Escherichia coli. J Membr Biol 76, 95-112.[CrossRef]
    [Google Scholar]
  12. Kay, W. W. ( 1978; ). Transport of carboxylic acids. In Bacterial Transport , pp. 385-411. Edited by B. P. Rosen. New York:Marcel Dekker.
  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 Bacteriol 182, 6374-6381.[CrossRef]
    [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 Bacteriol 183, 5862-5869.[CrossRef]
    [Google Scholar]
  15. Kunst, F., Ogasawara, N., Moszer, I. & 48 other authors ( 1997; ). The complete genome sequence of the Gram-positive bacterium Bacillus subtilis. Nature 390, 249–256.[CrossRef]
    [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 Biochem 220, 469-475.[CrossRef]
    [Google Scholar]
  17. Martell, A. E. & Smith, R. M. (1977). Critical Stability Constants, vol. 3: Other Organic Ligands. New York: Plenum.
  18. McKillen, M. N., Willecke, K. & Pardee, A. B. ( 1972; ). Citrate transport by Bacillus subtilis. In The Molecular Basis of Biological Transport , pp. 249-70. Edited by J. F. Woessner & F. Huijung. New York:Academic Press.
  19. Miller, J. H. (1972). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  20. Somers, J. M., Sweet, G. D. & Kay, W. W. ( 1981; ). Fluorocitrate resistant tricarboxylate transport mutants of Salmonella typhimurium. Mol Gen Genet 181, 338-345.[CrossRef]
    [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 Chem 259, 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 Bacteriol 174, 4893-4898.
     [Google Scholar]
  23. Warner, J. B. & Lolkema, J. S. ( 2002; ). LacZ-promoter fusions: the effect of growth. Microbiology 148, 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 Bacteriol 182, 6099-6105.[CrossRef]
    [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 Chem 275, 30287-30292.[CrossRef]
    [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 Bacteriol 170, 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 Microbiol 37, 898-912.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-11-3405
Loading
Growth of Bacillus subtilis on citrate and isocitrate is supported by the Mg2+–citrate transporter CitM
Microbiology 148, 3405 (2002); https://doi.org/10.1099/00221287-148-11-3405
/content/journal/micro/10.1099/00221287-148-11-3405
/content/journal/micro/10.1099/00221287-148-11-3405
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error