1887

Abstract

Quorum sensing is a signalling mechanism that controls diverse biological functions, including virulence, via -acylhomoserine lactone (AHL) signal molecules in Gram-negative bacteria. With the aim of isolating strains or enzymes capable of blocking quorum sensing by inactivating AHL, bacteria were screened for AHL degradation by their ability to utilize -3-oxohexanoyl--homoserine lactone (OHHL) as the sole carbon source. Among four isolates, strain IBN110, identified as sp., was found to grow rapidly on OHHL, and to degrade various AHLs with different lengths and acyl side-chain substitutions. Co-culture of sp. IBN110 and the plant pathogen significantly reduced both the AHL amount and pectate lyase activity in co-culture medium, suggesting the possibility of applying sp. IBN110 in the control of AHL-producing pathogenic bacteria. The gene from sp. IBN110 encoding the enzyme catalysing AHL degradation was cloned, and found to encode a protein of 273 amino acids. A mass spectrometry analysis showed that AhlD probably hydrolyses the lactone ring of -3-hexanoyl--homoserine lactone, indicating that AhlD is an -acylhomoserine lactonase (AHLase). A comparison of AhlD with other known AHL-degrading enzymes, sp. 240B1 AiiA, a subsp. AiiA homologue and AttM, revealed 25, 26 and 21 % overall identities, respectively, in the deduced amino acid sequences. Although these identities were relatively low, the HXDH≈H≈D motif was conserved in all the AHLases, suggesting that this motif is essential for AHLase activity. From a genome database search based on the conserved motif, putative AhlD-like lactonase genes were found in several other bacteria, and AHL-degrading activities were observed in and . Furthermore, it was verified that , an homologue, encodes an AHL-degrading enzyme in . Accordingly, the current results suggest the possibility that AhlDlike AHLases could exist in many other micro-organisms.

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2003-06-01
2019-09-23
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References

  1. Bainton, N. J., Stead, P., Chhabra, S. R., Bycroft, B. W., Salmond, G. P., Stewart, G. S. & Williams, P. ( 1992; ). N-(3-oxohexanoyl)-l-homoserine lactone regulates carbapenem antibiotic production in Erwinia carotovora. Biochem J 288, 997–1004.
    [Google Scholar]
  2. Chen, C. M. & Tomasek, P. H. ( 1991; ). 3,4-Dihydroxyxanthone dioxygenase from Arthrobacter sp. strain GFB100. Appl Environ Microbiol 57, 2217–2222.
    [Google Scholar]
  3. Cook, D. M., Li, P. L., Ruchaud, F., Padden, S. & Farrand, S. K. ( 1997; ). Ti plasmid conjugation is independent of vir: reconstitution of the tra functions from pTiC58 as a binary system. J Bacteriol 179, 1291–1297.
    [Google Scholar]
  4. Davies, D. G., Parsek, M. R., Pearson, J. P., Iglewski, B. H., Costerton, J. W. & Greenberg, E. P. ( 1998; ). The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280, 295–298.[CrossRef]
    [Google Scholar]
  5. De Kievit, T. R. & Iglewski, B. H. ( 2000; ). Bacterial quorum sensing in pathogenic relationships. Infect Immun 68, 4839–4849.[CrossRef]
    [Google Scholar]
  6. Dong, Y. H., Xu, J. L., Li, X. Z. & Zhang, L. H. ( 2000; ). AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. Proc Natl Acad Sci U S A 97, 3526–3531.[CrossRef]
    [Google Scholar]
  7. Dong, Y. H., Wang, L. H., Xu, J. L., Zhang, H. B., Zhang, X. F. & Zhang, L. H. ( 2001; ). Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase. Nature 411, 813–817.[CrossRef]
    [Google Scholar]
  8. Dong, Y. H., Gusti, A. R., Zhang, Q., Xu, J. L. & Zhang, L. H. ( 2002; ). Identification of quorum-quenching N-acyl homoserine lactonases from Bacillus species. Appl Environ Microbiol 68, 1754–1759.[CrossRef]
    [Google Scholar]
  9. Eberl, L., Winson, M. K., Sternberg, C. & 7 other authors ( 1996; ). Involvement of N-acyl-l-homoserine lactone autoinducers in controlling the multicellular behaviour of Serratia liquefaciens. Mol Microbiol 20, 127–136.[CrossRef]
    [Google Scholar]
  10. Finch, R. G., Pritchard, D. I., Bycroft, B. W., Williams, P. & Stewart, G. S. ( 1998; ). Quorum sensing: a novel target for anti-infective therapy. J Antimicrob Chemother 42, 569–571.[CrossRef]
    [Google Scholar]
  11. Flagan, S., Ching, W. K. & Leadbetter, J. R. ( 2003; ). Arthrobacter strain VAI-A utilizes acyl-homoserine lactone inactivation products and stimulates quorum signal biodegradation by Variovorax paradoxus. Appl Environ Microbiol 69, 909–916.[CrossRef]
    [Google Scholar]
  12. Hastings, J. W. & Nealson, K. H. ( 1977; ). Bacterial bioluminescence. Annu Rev Microbiol 31, 549–595.[CrossRef]
    [Google Scholar]
  13. Jones, S., Yu, B., Bainton, N. J. & 7 other authors ( 1993; ). The lux autoinducer regulates the production of exoenzyme virulence determinants in Erwinia carotovora and Pseudomonas aeruginosa. EMBO J 12, 2477–2482.
    [Google Scholar]
  14. Koch, C., Schumann, P. & Stackebrandt, E. ( 1995; ). Reclassification of Micrococcus agilis (Ali-Cohen 1889) to the genus Arthrobacter as Arthrobacter agilis comb. nov. and emendation of the genus Arthrobacter. Int J Syst Bacteriol 45, 837–839.[CrossRef]
    [Google Scholar]
  15. Latifi, A., Winson, M. K., Foglino, M., Bycroft, B. W., Stewart, G. S., Lazdunski, A. & Williams, P. ( 1995; ). Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PAO1. Mol Microbiol 17, 333–343.[CrossRef]
    [Google Scholar]
  16. Laurent, P., Buchon, L., Guespin-Michel, J. F. & Orange, N. ( 2000; ). Production of pectate lyases and cellulases by Chryseomonas luteola strain MFCL0 depends on the growth temperature and the nature of the culture medium: evidence for two critical temperatures. Appl Environ Microbiol 66, 1538–1543.[CrossRef]
    [Google Scholar]
  17. Leadbetter, J. R. & Greenberg, E. P. ( 2000; ). Metabolism of acyl-homoserine lactone quorum-sensing signals by Variovorax paradoxus. J Bacteriol 182, 6921–6926.[CrossRef]
    [Google Scholar]
  18. Lee, S. J., Park, S. Y., Lee, J. J., Yum, D. Y., Koo, B. T. & Lee, J. K. ( 2002; ). Genes encoding the N-acyl homoserine lactone-degrading enzyme are widespread in many subspecies of Bacillus thuringiensis. Appl Environ Microbiol 68, 3919–3924.[CrossRef]
    [Google Scholar]
  19. Lin, Y. H., Xu, J. L., Hu, J., Wang, L. H., Ong, S. L., Leadbetter, J. R. & Zhang, L. H. ( 2003; ). Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a novel and potent class of quorum-quenching enzymes. Mol Microbiol 47, 849–860.[CrossRef]
    [Google Scholar]
  20. Loh, J., Carlson, R. W., York, W. S. & Stacey, G. ( 2002; ). Bradyoxetin, a unique chemical signal involved in symbiotic gene regulation. Proc Natl Acad Sci U S A 99, 14446–14451.[CrossRef]
    [Google Scholar]
  21. Maidak, B. L., Cole, J. R., Parker, C. T., Jr & 11 other authors ( 1999; ). A new version of the RDP (Ribosomal Database Project). Nucleic Acids Res 27, 171–173.[CrossRef]
    [Google Scholar]
  22. Nealson, K. H. & Hastings, J. W. ( 1979; ). Bacterial bioluminescence: its control and ecological significance. Microbiol Rev 43, 496–518.
    [Google Scholar]
  23. Pearson, J. P., Gray, K. M., Passador, L., Tucker, K. D., Eberhard, A., Iglewski, B. H. & Greenberg, E. P. ( 1994; ). Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. Proc Natl Acad Sci U S A 91, 197–201.[CrossRef]
    [Google Scholar]
  24. Pearson, J. P., Pesci, E. C. & Iglewski, B. H. ( 1997; ). Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems in control of elastase and rhamnolipid biosynthesis genes. J Bacteriol 179, 5756–5767.
    [Google Scholar]
  25. Pierson, L. S. I., Keppenne, V. D. & Wood, D. W. ( 1994; ). Phenazine antibiotic biosynthesis in Pseudomonas aureofaciens 30-84 is regulated by PhzR in response to cell density. J Bacteriol 176, 3966–3974.
    [Google Scholar]
  26. Piper, K. R., Beck von Bodman, S. & Farrand, S. K. ( 1993; ). Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction. Nature 362, 448–450.[CrossRef]
    [Google Scholar]
  27. Salmond, G. P., Bycroft, B. W., Stewart, G. S. & Williams, P. ( 1995; ). The bacterial ‘enigma’: cracking the code of cell-cell communication. Mol Microbiol 16, 615–624.[CrossRef]
    [Google Scholar]
  28. Sayler, G. S., Hooper, S. W., Layton, A. C. & King., J. M. H. ( 1990; ). Catabolic plasmids of environmental and ecological significance. Microb Ecol 19, 1–20.[CrossRef]
    [Google Scholar]
  29. Swift, S., Throup, J. P., Williams, P., Salmond, G. P. & Stewart, G. S. ( 1996; ). Quorum sensing: a population-density component in the determination of bacterial phenotype. Trends Biochem Sci 21, 214–219.[CrossRef]
    [Google Scholar]
  30. Throup, J. P., Camara, M., Briggs, G. S., Winson, M. K., Chhabra, S. R., Bycroft, B. W., Williams, P. & Stewart, G. S. ( 1995; ). Characterisation of the yenI/yenR locus from Yersinia enterocolitica mediating the synthesis of two N-acylhomoserine lactone signal molecules. Mol Microbiol 17, 345–356.[CrossRef]
    [Google Scholar]
  31. Vallee, B. & Galdes, A. ( 1984; ). The metallobiochemistry of zinc enzymes. Adv Enzymol Relat Areas Mol Biol 56, 283–430.
    [Google Scholar]
  32. van Waasbergen, L. G., Balkwill, D. L., Crocker, F. H., Bjornstad, B. N. & Miller, R. V. ( 2000; ). Genetic diversity among Arthrobacter species collected across a heterogeneous series of terrestrial deep-subsurface sediments as determined on the basis of 16S rRNA and recA gene sequences. Appl Environ Microbiol 66, 3454–3463.[CrossRef]
    [Google Scholar]
  33. Whitehead, N. A., Welch, M. & Salmond, G. P. ( 2001; ). Silencing the majority. Nat Biotechnol 19, 735–736.[CrossRef]
    [Google Scholar]
  34. Yates, E. A., Philipp, B., Buckley, C. & 8 other authors ( 2002; ). N-Acylhomoserine lactones undergo lactonolysis in a pH-, temperature-, and acyl chain length-dependent manner during growth of Yersinia pseudotuberculosis and Pseudomonas aeruginosa. Infect Immun 70, 5635–5646.[CrossRef]
    [Google Scholar]
  35. Zhang, L., Murphy, P. J., Kerr, A. & Tate, M. E. ( 1993; ). Agrobacterium conjugation and gene regulation by N-acyl-l-homoserine lactones. Nature 362, 446–448.[CrossRef]
    [Google Scholar]
  36. Zhang, H. B., Wang, L. H. & Zhang, L. H. ( 2002; ). Genetic control of quorum-sensing signal turnover in Agrobacterium tumefaciens. Proc Natl Acad Sci U S A 99, 4638–4643.[CrossRef]
    [Google Scholar]
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