Advertisement

A novel medium for the isolation of N-acylhomoserine lactone-degrading bacteria

  • Kok-Gan Chan
  • Wai-Fong Yin
  • Choon-Kook Sam
  • Chong-Lek Koh
Original Paper

Abstract

A novel chemically defined medium, named KG medium, supplemented with N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL), an acylhomoserine lactone (AHL) used as signalling molecules in Gram-negative bacterial cell-to-cell communication, as the sole source of carbon and nitrogen, was designed and successfully used for the enrichment and isolation of AHL-degrading bacteria. A 3-oxo-C6-HSL-degrading bacterium, 13sw7, was isolated from sewage after six enrichment transfers in the 3-oxo-C6-HSL-containing KG medium. On the basis of the almost complete 16S ribosomal DNA sequence, isolate 13sw7 was clustered with unculturable β-proteobacteria. This study indicates that the AHL-containing KG medium is effective in isolating AHL-degrading bacteria, including those previously considered unculturable, from environmental sources. To the best of our knowledge, this is the first documentation of the isolation of an AHL-degrading proteobacterium from sewage.

Keywords

KG medium N-3-oxo-hexanoylhomoserine lactone Proteobacterium Quorum sensing Quorum quenching Sewage 

Notes

Acknowledgments

K. G. C. thanks the University of Malaya for a Ph.D. fellowship and the Commonwealth Scholarship Commission, UK, for a Commonwealth Split-site Ph.D. Scholarship. We thank Paul Williams (Nottingham, UK) for the biosensor and AHL. We thank Joel Low Z. B. for technical assistance in DNA sequencing and phylogenetic analysis. This research was supported by the Malaysia Toray Science Foundation (MTSF1013-2007A), Research University Grants (University of Malaya, grant no. FR146/2007A, TA048/2008A), and two Vote F research grants from the University of Malaya to K. G. C.

References

  1. 1.
    Chan KG, Tiew SZ, Ng CC (2007) Rapid isolation method of soil bacilli and screening of their quorum quenching activity. As Pac J Mol Biol Biotech 15:153–156Google Scholar
  2. 2.
    Dewhirst FE, Chien CC, Paster BJ, Ericson RL, Orcutt RP, Schauer DB, Fox JG (1999) Phylogeny of the defined murine microbiota: Altered Schaedler fora. Appl Environ Microbiol 65:3287–3292PubMedGoogle Scholar
  3. 3.
    Dong YH, Xu JL, Li XZ, Zhang LH (2000) AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. Proc Natl Acad Sci USA 97:3526–3531. doi: 10.1073/pnas.060023897 PubMedCrossRefGoogle Scholar
  4. 4.
    Dong YH, Wang LH, Xu JL, Zhang HB, Zhang XF, Zhang LH (2001) Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase. Nature 411:813–817. doi: 10.1038/35081101 PubMedCrossRefGoogle Scholar
  5. 5.
    Fuqua WC, Greenberg EP (1998) Self perception in bacteria: quorum sensing with acylated homoserine lactones. Curr Opin Microbiol 1:183–189. doi: 10.1016/S1369-5274(98)80009-X PubMedCrossRefGoogle Scholar
  6. 6.
    Fuqua WC, Winans SC, Greenberg EP (1994) Quorum sensing in bacteria—the LuxR-LuxI family of cell density-responsive transcriptional regulators. J Bacteriol 176:269–275PubMedGoogle Scholar
  7. 7.
    Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML, Pace NR (1985) Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci USA 82:6955–6959. doi: 10.1073/pnas.82.20.6955 PubMedCrossRefGoogle Scholar
  8. 8.
    Leadbetter JR, Greenberg EP (2000) Metabolism of acyl-homoserine lactone quorum-sensing signals by Variovorax paradoxus. J Bacteriol 182:6921–6926. doi: 10.1128/JB.182.24.6921-6926.2000 PubMedCrossRefGoogle Scholar
  9. 9.
    McClean KH, Winson MK, Fish L, Taylor A, Chhabra SR, Camara M, Daykin M, Lamb JH, Swift S, Bycroft BW, Stewart GSAB, Williams P (1997) Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143:3703–3711PubMedCrossRefGoogle Scholar
  10. 10.
    Sio CF, Otten LG, Cool RH, Diggle SP, Braun PG, Bos R, Daykin M, Camara M, Williams P, Quax WJ (2006) Quorum quenching by an N-acyl-homoserine lactone acylase from Pseudomonas aeruginosa PAO1. Infect Immun 74:1673–1682. doi: 10.1128/IAI.74.3.1673-1682.2006 PubMedCrossRefGoogle Scholar
  11. 11.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599. doi: 10.1093/molbev/msm092 PubMedCrossRefGoogle Scholar
  12. 12.
    Uroz S, Angelo-Picard C, Carlier A, Elasri M, Sicot C, Petit A, Oger P, Faure D, Dessaux Y (2003) Novel bacteria degrading N-acylhomoserine lactones and their use as quenchers of quorum-sensing-regulated functions of plant-pathogenic bacteria. Microbiology 149:1981–1989. doi: 10.1099/mic.0.26375-0 PubMedCrossRefGoogle Scholar
  13. 13.
    Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703PubMedGoogle Scholar
  14. 14.
    Whitehead NA, Barnard AML, Slater H, Simpson NJL, Salmond GPC (2001) Quorum-sensing in gram-negative bacteria. FEMS Microbiol Rev 25:365–404. doi: 10.1111/j.1574-6976.2001.tb00583.x PubMedCrossRefGoogle Scholar
  15. 15.
    Williams P (2002) Quorum sensing: an emerging target for antibacterial chemotherapy? Expert Opin Ther Targets 6:1–8. doi: 10.1517/14728222.6.3.257 CrossRefGoogle Scholar
  16. 16.
    Williams P, Camara M, Hardman A, Swift S, Milton D, Hope VJ, Winzer K, Middleton B, Pritchard DI, Bycroft BW (2000) Quorum sensing and the population-dependent control of virulence. Philos Trans R Soc Lond B Biol Sci 355:667–680. doi: 10.1098/rstb.2000.0607 PubMedCrossRefGoogle Scholar
  17. 17.
    Yates EA, Philipp B, Buckley C, Atkinson S, Chhabra SR, Sockett RE, Goldner M, Dessaux Y, Camara M, Smith H, Williams P (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. doi: 10.1128/IAI.70.10.5635-5646.2002 PubMedCrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology 2008

Authors and Affiliations

  • Kok-Gan Chan
    • 1
  • Wai-Fong Yin
    • 1
  • Choon-Kook Sam
    • 2
  • Chong-Lek Koh
    • 2
  1. 1.Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia
  2. 2.Natural Sciences and Science Education AG, National Institute of EducationNanyang Technological UniversitySingaporeSingapore

Personalised recommendations