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Antonie van Leeuwenhoek

, Volume 108, Issue 2, pp 491–504 | Cite as

Isoprenyl caffeate, a major compound in manuka propolis, is a quorum-sensing inhibitor in Chromobacterium violaceum

  • Adrian Tandhyka Gemiarto
  • Nathaniel Nyakaat Ninyio
  • Siew Wei Lee
  • Joko Logis
  • Ayesha Fatima
  • Eric Wei Chiang Chan
  • Crystale Siew Ying Lim
Original Paper

Abstract

The emergence of antibiotic-resistant bacterial pathogens, especially Gram-negative bacteria, has driven investigations into suppressing bacterial virulence via quorum sensing (QS) inhibition strategies instead of bactericidal and bacteriostatic approaches. Here, we investigated several bee products for potential compound(s) that exhibit significant QS inhibitory (QSI) properties at the phenotypic and molecular levels in Chromobacterium violaceum ATCC 12472 as a model organism. Manuka propolis produced the strongest violacein inhibition on C. violaceum lawn agar, while bee pollen had no detectable QSI activity and honey had bactericidal activity. Fractionated manuka propolis (pooled fraction 5 or PF5) exhibited the largest violacein inhibition zone (24.5 ± 2.5 mm) at 1 mg dry weight per disc. In C. violaceum liquid cultures, at least 450 µg/ml of manuka propolis PF5 completely inhibited violacein production. Gene expression studies of the vioABCDE operon, involved in violacein biosynthesis, showed significant (≥two-fold) down-regulation of vioA, vioD and vioE in response to manuka propolis PF5. A potential QSI compound identified in manuka propolis PF5 is a hydroxycinnamic acid-derivative, isoprenyl caffeate, with a [M−H] of 247. Complete violacein inhibition in C. violaceum liquid cultures was achieved with at least 50 µg/ml of commercial isoprenyl caffeate. In silico docking experiments suggest that isoprenyl caffeate may act as an inhibitor of the violacein biosynthetic pathway by acting as a competitor for the FAD-binding pockets of VioD and VioA. Further studies on these compounds are warranted toward the development of anti-pathogenic drugs as adjuvants to conventional antibiotic treatments, especially in antibiotic-resistant bacterial infections.

Keywords

Quorum sensing inhibition Manuka propolis Isoprenyl caffeate vio operon Antibiotic resistance Chromobacterium violaceum 

Notes

Acknowledgments

This work was done with funding from UCSI University’s Faculty of Applied Sciences and the MAKNA Cancer Research Award 2012. We thank Prof. Tom Coenye for his constructive comments of the early findings of this study and helpful insights into the QSI activity of cinnamaldehyde and cinnamaldehyde derivatives. We also thank the Drug Design & Development Research Group (DDDRG) of the University of Malaya for use of the docking software.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Annapoorani A, Parameswari R, Pandian SK, Ravi AV (2012) Methods to determine antipathogenic potential of phenolic and flavonoid compounds against urinary pathogen Serratia marcescens. J Microbiol Methods 91:208–211PubMedCrossRefGoogle Scholar
  2. August PR, Grossman TH, Minor C, Draper MP, MacNeil IA, Pemberton JM, Call KM, Holt D, Osburne MS (2000) Sequence analysis and functional characterization of the violacein biosynthetic pathway from Chromobacterium violaceum. J Mol Microbiol Biotechnol 2:513–519PubMedGoogle Scholar
  3. Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The protein data bank. Nucleic Acids Res 28:235–242PubMedCentralPubMedCrossRefGoogle Scholar
  4. Brackman G, Defoirdt T, Miyamoto C, Bossier P, Van CS, Nelis H, Coenye T (2008) Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR. BMC Microbiol 8:149PubMedCentralPubMedCrossRefGoogle Scholar
  5. Brackman G, Celen S, Hillaert U, Van Calenbergh S, Cos P, Maes L, Nelis HJ, Coenye T (2011) Structure-activity relationship of cinnamaldehyde analogs as inhibitors of AI-2 based quorum sensing and their effect on virulence of Vibrio spp. PLoS One 6:e16084PubMedCentralPubMedCrossRefGoogle Scholar
  6. Brazilian National Project Consortium (2003) The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability. Proc Natl Acad Sci USA 100:11660–11665CrossRefGoogle Scholar
  7. Bulman Z, Lee P, Hudson AO, Savka MA (2011) A novel property of propolis (bee glue): anti-pathogenic activity by inhibition of N-acyl-homoserine lactone mediated signaling in bacteria. J Ethnopharmacol 138:788–797PubMedCrossRefGoogle Scholar
  8. Falcão SI, Vilas-Boas M, Estevinho LM, Barros C, Domingues MR, Cardoso SM (2010) Phenolic characterization of Northeast Portuguese propolis: usual and unusual compounds. Anal Bioanal Chem 396:887–897PubMedCrossRefGoogle Scholar
  9. Gouveia SC, Castilho PC (2012) Validation of a HPLC-DAD–ESI/MSn method for caffeoylquinic acids separation, quantification and identification in medicinal Helichrysum species from Macaronesia. Food Res Int 45:362–368CrossRefGoogle Scholar
  10. Hentzer M, Givskov M (2003) Pharmacological inhibition of quorum sensing for the treatment of chronic bacterial infections. J Clin Invest 112:1300–1307PubMedCentralPubMedCrossRefGoogle Scholar
  11. Hentzer M, Wu H, Andersen JB, Riedel K, Rasmussen TB, Bagge N, Kumar N, Schembri MA, Song Z, Kristoffersen P, Manefield M, Costerton JW, Molin S, Eberl L, Steinberg P, Kjelleberg S, Høiby N, Givskov M (2003) Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO J 22:3803–3815PubMedCentralPubMedCrossRefGoogle Scholar
  12. Lamberte LE, Cabrera EC, Rivera WL (2011) Activity of the ethanolic extract of propolis (EEP) as a potential inhibitor of quorum sensing-mediated pigment production in Chromobacterium violaceum and virulence factor production in Pseudomonas aeruginosa. Philipp Agric Sci 94:14–22Google Scholar
  13. Lee JH, Park JH, Kim JA, Neupane GP, Cho MH, Lee CS, Lee J (2011) Low concentrations of honey reduce biofilm formation, quorum sensing, and virulence in Escherichia coli O157:H7. Biofouling 27:1095–1104PubMedCrossRefGoogle Scholar
  14. Martinelli D, Grossmann G, Séquin U, Brandl H, Bachofen M (2004) Effects of natural and chemically synthesized furanones on quorum sensing in Chromobacterium violaceum. BMC Microbiol 4:25PubMedCentralPubMedCrossRefGoogle Scholar
  15. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45PubMedCentralPubMedCrossRefGoogle Scholar
  16. Rasmussen TB, Givskov M (2006) Quorum-sensing inhibitors as anti-pathogenic drugs. Int J Med Microbiol 296:149–161PubMedCrossRefGoogle Scholar
  17. Rasmussen TB, Bjarnsholt T, Skindersoe ME, Hentzer M, Kristoffersen P, Köte M, Nielsen J, Eberl L, Givskov M (2005) Screening for quorum-sensing inhibitors (QSI) by use of a novel genetic system, the QSI Selector. J Bacteriol 187:1799–1814PubMedCentralPubMedCrossRefGoogle Scholar
  18. Ryan KS, Balibar CJ, Turo KE, Walsh CT, Drennan CL (2008) The violacein biosynthetic enzyme VioE shares a fold with lipoprotein transporter proteins. J Biol Chem 283:6467–6475PubMedCrossRefGoogle Scholar
  19. Schwede T, Kopp J, Guex N, Peitsch M (2003) SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res 31:3381–3385PubMedCentralPubMedCrossRefGoogle Scholar
  20. Singh BN, Singh BR, Singh RL, Prakash D, Dhakarey R, Upadhyay G, Singh HB (2009) Oxidative DNA damage protective activity, antioxidant and anti-quorum sensing potentials of Moringa oleifera. Food Chem Toxicol 47:1109–1116PubMedCrossRefGoogle Scholar
  21. Slama TG (2008) Gram-negative antibiotic resistance: there is a price to pay. Crit Care 12(Suppl 10):1–7Google Scholar
  22. Souli M, Galani I, Giamarellou H (2008) Emergence of extensively drug-resistant and pandrug-resistant Gram-negative bacilli in Europe. Euro Surveill 13:19045PubMedGoogle Scholar
  23. Stauff DL, Bassler BL (2011) Quorum sensing in Chromobacterium violaceum: DNA recognition and gene regulation by the CviR receptor. J Bacteriol 193:3871–3878PubMedCentralPubMedCrossRefGoogle Scholar
  24. Taganna JC, Rivera WL (2008) Epigallocatechin gallate from Camellia sinensis L. (Kuntze) is a potential quorum sensing inhibitor in Chromobacterium violaceum. Sci Diliman 20:24–30Google Scholar
  25. Taganna JC, Quanico JP, Perono RM, Amor EC, Rivera WL (2011) Tannin-rich fraction from Terminalia catappa inhibits quorum sensing (QS) in Chromobacterium violaceum and the QS-controlled biofilm maturation and LasA staphylolytic activity in Pseudomonas aeruginosa. J Ethnopharmacol 134:865–871PubMedCrossRefGoogle Scholar
  26. Wagner VE, Gillis RJ, Iglewski BH (2004) Transcriptome analysis of quorum-sensing regulation and virulence factor expression in Pseudomonas aeruginosa. Vaccine 22(Suppl 1):S15–S20PubMedCrossRefGoogle Scholar
  27. Wallace AC, Laskowski RA, Thornton JM (1995) LIGPLOT: a program to generate schematic diagrams of protein-ligand interactions. Protein Eng 8:127–134PubMedCrossRefGoogle Scholar
  28. Wang R, Starkey M, Hazan R, Rahme LG (2012) Honey’s ability to counter bacterial infections arises from both bactericidal compounds and QS inhibition. Front Microbiol 2012:144Google Scholar
  29. Waters CM, Bassler BL (2005) Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol 21:319–346PubMedCrossRefGoogle Scholar
  30. Winser K, Williams P (2001) Quorum sensing and the regulation of virulence gene expression in pathogenic bacteria. Int J Med Microbiol 291:131–143CrossRefGoogle Scholar
  31. Wu G, Robertson DH, Brooks CL 3rd, Vieth M (2003) Detailed analysis of grid-based molecular docking: a case study of CDOCKER-A CHARMm-based MD docking algorithm. J Comput Chem 24:1549–1562PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Adrian Tandhyka Gemiarto
    • 1
    • 4
  • Nathaniel Nyakaat Ninyio
    • 1
    • 5
  • Siew Wei Lee
    • 1
  • Joko Logis
    • 1
  • Ayesha Fatima
    • 3
  • Eric Wei Chiang Chan
    • 2
  • Crystale Siew Ying Lim
    • 1
  1. 1.Department of Biotechnology, Faculty of Applied SciencesUCSI UniversityCheras, Kuala LumpurMalaysia
  2. 2.Department of Food Science and Nutrition, Faculty of Applied SciencesUCSI UniversityCheras, Kuala LumpurMalaysia
  3. 3.Faculty of Pharmaceutical SciencesUCSI UniversityKuala LumpurMalaysia
  4. 4.Comparative Genomics CentreJames Cook UniversityTownsvilleAustralia
  5. 5.Department of Microbiology, Faculty of ScienceKaduna State UniversityKadunaNigeria

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