Skip to main content
SpringerLink
Log in

Two novel synthetic peptides inhibit quorum sensing-dependent biofilm formation and some virulence factors in Pseudomonas aeruginosa PAO1

  • Microbial Pathogenesis and Host-Microbe Interaction
  • Published:
Journal of Microbiology Aims and scope Submit manuscript

Abstract

Quorum sensing (QS) regulates virulence factor expression in Pseudomonas aeruginosa. Inhibiting the QS-controlled virulence factors without inhibiting the growth of P. aeruginosa is a promising approach for overcoming the widespread resistance of P. aeruginosa. This study was proposed to investigate the effects of two novel synthetic peptides on the biofilm development and virulence factor production of P. aeruginosa. The tested strain was P. aeruginosa PAO1. The results indicated that both of the synthetic peptides (LIVRHK and LIVRRK) inhibited (P < 0.05) the formation of biofilms and the production of virulence factors, including pyocyanin, protease, and rhamnolipids, without inhibiting the growth of PAO1. Additionally, we detected transcriptional changes related to QS and found a significant reduction in the levels of gene expression of lasI, lasR, rhlI, and rhlR. This study demonstrates that LIVRRK and LIVRHK are novel synthetic peptides that can act as potent inhibitors of QS-regulated virulence factors in P. aeruginosa. Moreover, these synthetic peptides have potential applications in the treatment of biofilmrelated diseases. Both peptides may be able to control chronic infections and biofilm-associated problems of P. aeruginosa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Abdel-Mawgoud, A.M., Hausmann, R., Lépine, F., Müller, M.M., and Déziel, E. 2011. Rhamnolipids: detection, analysis, biosynthesis, genetic regulation, and bioengineering of production, pp. 13–55. In Biosurfactants, Springer, Berlin, Hdidelberg, Germany.

    Chapter  Google Scholar 

  • Andrejko, M., Zdybicka-Barabas, A., Janczarek, M., and Cytryńska, M. 2013. Three Pseudomonas aeruginosa strains with different protease profiles. Acta Biochim. Pol. 60, 83–90.

    Article  PubMed  Google Scholar 

  • Ayora, S. and Götz, F. 1994. Genetic and biochemical properties of an extracellular neutral metalloprotease from Staphylococcus hyicus subsp. hyicus. Mol. Gen. Genet. 242, 421.

    CAS  PubMed  Google Scholar 

  • Azghani, A.O., Miller, E.J., and Peterson, B.T. 2000. Virulence factors from Pseudomonas aeruginosa increase lung epithelial permeability. Lung 178, 261–269.

    Article  CAS  PubMed  Google Scholar 

  • Bala, A., Kumar, R., and Harjai, K. 2011. Inhibition of quorum sensing in Pseudomonas aeruginosa by azithromycin and its effectiveness in urinary tract infections. J. Med. Microbiol. 60, 300–306.

    Article  CAS  PubMed  Google Scholar 

  • Balaji, K., Thenmozhi, R., and Pandian, S.K. 2013. Effect of subinhibitory concentrations of fluoroquinolones on biofilm production by clinical isolates of Streptococcus pyogenes. Indian J. Med. Res. 137, 963.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Brogden, K.A. 2005. Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria? Nat. Rev. Microbiol. 3, 238.

    Article  CAS  PubMed  Google Scholar 

  • Cherkasov, A., Hilpert, K., Jenssen, H., Fjell, C.D., Waldbrook, M., Mullaly, S.C., Volkmer, R., and Hancock, R.E.W. 2008. Use of artificial intelligence in the design of small peptide antibiotics effective against a broad spectrum of highly antibiotic-resistant superbugs. ACS Chem. Biol. 4, 65–74.

    Article  CAS  Google Scholar 

  • CLSI (Clinical and Laboratory Standards Institute). 2012. Performance standards for antimicrobial disk susceptibility tests; Approved standards. Wayne, PA: M100–S22.

    Google Scholar 

  • De Kievit, T.R. 2009. Quorum sensing in Pseudomonas aeruginosa biofilms. Environ. Microbiol. 11, 279–288.

    Article  CAS  PubMed  Google Scholar 

  • de la Fuente-Núñez, C., Cardoso, M.H., de Souza Cândido, E., Franco, O.L., and Hancock, R.E.W. 2016. Synthetic antibiofilm peptides. BBA-Biomembranes 1858, 1061–1069.

    Article  CAS  PubMed  Google Scholar 

  • de la Fuente-Núñez, C., Korolik, V., Bains, M., Nguyen, U., Breidenstein, E.B.M., Horsman, S., Lewenza, S., Burrows, L., and Hancock, R.E.W. 2012. Inhibition of bacterial biofilm formation and swarming motility by a small synthetic cationic peptide. Antimicrob. Agents Chemother. 56, 2696–2704.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Déziel, E., Lépine, F., Milot, S., and Villemur, R. 2000. Mass spectrometry monitoring of rhamnolipids from a growing culture of Pseudomonas aeruginosa strain 57RP. Biochim. Biophys. Acta (BBA)-Mol. Cell Biol. Lipids 1485, 145–152.

    Article  Google Scholar 

  • Essar, D.W., Eberly, L., Hadero, A., and Crawford, I.P. 1990. Identification and characterization of genes for a second anthranilate synthase in Pseudomonas aeruginosa: interchangeability of the two anthranilate synthases and evolutionary implications. J. Bacteriol. 172, 884–900.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fjell, C.D., Hiss, J.A., Hancock, R.E.W., and Schneider, G. 2012. Designing antimicrobial peptides: form follows function. Nat. Rev. Drug Discov. 11, 37.

    Article  CAS  Google Scholar 

  • Fuqua, C. and Greenberg, E.P. 2002. Signalling: listening in on bacteria: acyl-homoserine lactone signalling. Nat. Rev. Mol. Cell Biol. 3, 685.

    Article  CAS  PubMed  Google Scholar 

  • Gonzâlez-Lamothe, R., Mitchell, G., Gattuso, M., Diarra, M.S., Malouin, F., and Bouarab, K. 2009. Plant antimicrobial agents and their effects on plant and human pathogens. Int. J. Mol. Sci. 10, 3400–3419.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta, S., Sharma, A.K., Jaiswal, S.K., and Sharma, V.K. 2016. Prediction of biofilm inhibiting peptides: an in silico approach. Front. Microbiol. 7, 949.

    PubMed  PubMed Central  Google Scholar 

  • Hancock, R.E.W. and Sahl, H.G. 2006. Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat. Biotechnol. 24, 1551.

    Article  CAS  PubMed  Google Scholar 

  • Haney, E.F., Brito-Sânchez, Y., Trimble, M.J., Mansour, S.C., Cherkasov, A., and Hancock, R.E.W. 2018. Computer-aided discovery of peptides that specifically attack bacterial biofilms. Sci. Rep. 8, 1871.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Haney, E.F., Mansour, S.C., Hilchie, A.L., de la Fuente-Núñez, C., and Hancock, R.E.W. 2015. High throughput screening methods for assessing antibiofilm and immunomodulatory activities of synthetic peptides. Peptides 71, 276–285.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • He, J. and Furmanski, P. 1995. Sequence specificity and transcriptional activation in the binding of lactoferrin to DNA. Nature 373, 721–724.

    Article  CAS  PubMed  Google Scholar 

  • Hentzer, M., Riedel, K., Rasmussen, T.B., Heydorn, A., Andersen, J.B., Parsek, M.R., Rice, S.A., Eberl, L., Molin, S., Hoiby, N., et al. 2002. Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound. Microbiology 148, 87–102.

    Article  CAS  PubMed  Google Scholar 

  • Høiby, N. 1994. Diffuse panbronchiolitis and cystic fibrosis: East meets West. Thorax 49, 531.

    Article  PubMed  PubMed Central  Google Scholar 

  • Jensen, P.Ø., Bjarnsholt, T., Phipps, R., Rasmussen, T.B., Calum, H., Christoffersen, L., Moser, C., Williams, P., Pressler, T., and Givskov, M. 2007. Rapid necrotic killing of polymorphonuclear leukocytes is caused by quorum-sensing-controlled production of rhamnolipid by Pseudomonas aeruginosa. Microbiology 153, 1329–1338.

    Article  CAS  PubMed  Google Scholar 

  • Köhler, T., Guanella, R., Carlet, J., and Van Delden, C. 2010. Quorum sensing-dependent virulence during Pseudomonas aeruginosa colonization and pneumonia in mechanically ventilated patients. Thorax 65, 703–710.

    Article  PubMed  Google Scholar 

  • Krishnan, T., Yin, W.F., and Chan, K.G. 2012. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa PAO1 by Ayurveda spice clove (Syzygium aromaticum) bud extract. Sensors 12, 4016–4030.

    Article  CAS  PubMed  Google Scholar 

  • Lieberman, D. and Lieberman, D. 2003. Pseudomonal infections in patients with COPD. Am. J. Respir. Med. 2, 459–468.

    Article  PubMed  Google Scholar 

  • Lyczak, J.B., Cannon, C.L., and Pier, G.B. 2002. Lung infections associated with cystic fibrosis. Clin. Microbiol. Rev. 15, 194–222.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McKnight, S.L., Iglewski, B.H., and Pesci, E.C. 2000. The Pseudomonas quinolone signal regulates rhl quorum sensing in Pseudomonas aeruginosa. J. Bacteriol. 182, 2702–2708.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Melander, R.J. and Melander, C. 2017. The challenge of overcoming antibiotic resistance: an adjuvant approach? ACS Infect. Dis. 3, 559–563.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Morita, Y., Tomida, J., and Kawamura, Y. 2014. Responses of Pseudomonas aeruginosa to antimicrobials. Front. Microbiol. 4, 422.

    Article  PubMed  PubMed Central  Google Scholar 

  • Nalca, Y., Jänsch, L., Bredenbruch, F., Geffers, R., Buer, J., and Häussler, S. 2006. Quorum-sensing antagonistic activities of azithromycin in Pseudomonas aeruginosa PAO1: a global approach. Antimicrob. Agents Chemother. 50, 1680–1688.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nazzaro, F., Fratianni, F., and Coppola, R. 2013. Quorum sensing and phytochemicals. Int. J. Mol. Sci. 14, 12607–12619.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ochsner, U.A., Fiechter, A., and Reiser, J. 1994. Isolation, characterization, and expression in Escherichia coli of the Pseudomonas aeruginosa rhlAB genes encoding a rhamnosyltransferase involved in rhamnolipid biosurfactant synthesis. J. Biol. Chem. 269, 19787–19795.

    CAS  PubMed  Google Scholar 

  • Ouyang, J., Sun, F., Feng, W., Sun, Y., Qiu, X., Xiong, L., Liu, Y., and Chen, Y. 2016. Quercetin is an effective inhibitor of quorum sensing, biofilm formation and virulence factors in Pseudomonas aeruginosa. J. Appl. Microbiol. 120, 966–974.

    Article  CAS  PubMed  Google Scholar 

  • Overhage, J., Campisano, A., Bains, M., Torfs, E.C.W., Rehm, B.H.A., and Hancock, R.E.W. 2008. Human host defense peptide LL-37 prevents bacterial biofilm formation. Infect. Immun. 76, 4176–4182.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pesci, E.C., Milbank, J.B.J., Pearson, J.P., McKnight, S., Kende, A.S., Greenberg, E.P., and Iglewski, B.H. 1999. Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 96, 11229–11234.

    Article  CAS  PubMed  Google Scholar 

  • Powers, J.P.S., Martin, M.M., Goosney, D.L., and Hancock, R.E.W. 2006. The antimicrobial peptide polyphemusin localizes to the cytoplasm of Escherichia coli following treatment. Antimicrob. Agents Chemother. 50, 1522–1524.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rossolini, G.M. and Mantengoli, E. 2005. Treatment and control of severe infections caused by multiresistant Pseudomonas aeruginosa. Clin. Microbiol. Infect. 11, 17–32.

    Article  CAS  PubMed  Google Scholar 

  • Sarkar, R., Chaudhary, S.K., Sharma, A., Yadav, K.K., Nema, N.K., Sekhoacha, M., Karmakar, S., Braga, F.C., Matsabisa, M.G., and Mukherjee, P.K. 2014. Anti-biofilm activity of Marula-A study with the standardized bark extract. J. Ethnopharmacol. 154, 170–175.

    Article  PubMed  Google Scholar 

  • Savli, H., Karadenizli, A., Kolayli, F., Gundes, S., Ozbek, U., and Vahaboglu, H. 2003. Expression stability of six housekeeping genes: a proposal for resistance gene quantification studies of Pseudomonas aeruginosa by real-time quantitative RT-PCR. J. Med. Microbiol. 52, 403–408.

    Article  CAS  PubMed  Google Scholar 

  • Schuster, M. and Greenberg, E.P. 2006. A network of networks: quorum-sensing gene regulation in Pseudomonas aeruginosa. Int. J. Med. Microbiol. 296, 73–81.

    Article  CAS  PubMed  Google Scholar 

  • Sharma, G., Rao, S., Bansal, A., Dang, S., Gupta, S., and Gabrani, R. 2014. Pseudomonas aeruginosa biofilm: potential therapeutic targets. Biologicals 42, 1–7.

    Article  CAS  PubMed  Google Scholar 

  • Smith, R.S. and Iglewski, B.H. 2003. Pseudomonas aeruginosa quorum-sensing systems and virulence. Curr. Opin. Microbiol. 6, 56–60.

    Article  CAS  PubMed  Google Scholar 

  • Stepanović, S., Vuković, D., Hola, V., Bonaventura, G.D.I., Djukić, S., Ćirković, I., and Ruzicka, F. 2007. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 115, 891–899.

    Article  PubMed  Google Scholar 

  • Sui, S.J.H., Lo, R., Fernandes, A.R., Caulfield, M.D.G., Lerman, J.A., Xie, L., Bourne, P.E., Baillie, D.L., and Brinkman, F.S.L. 2012. Raloxifene attenuates Pseudomonas aeruginosa pyocyanin production and virulence. Int. J. Antimicrob. Agents 40, 246–251.

    Article  CAS  Google Scholar 

  • Tateda, K., Comte, R., Pechere, J.C., Köhler, T., Yamaguchi, K., and Van Delden, C. 2001. Azithromycin inhibits quorum sensing in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 45, 1930–1933.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Venturi, V. 2006. Regulation of quorum sensing in Pseudomonas. FEMS Microbiol. Rev. 30, 274–291.

    Article  CAS  PubMed  Google Scholar 

  • Vijayaraghavan, P. and Vincent, S.G.P. 2013. A simple method for the detection of protease activity on agar plates using bromocresolgreen dye. J. Biochem. Technol. 4, 628–630.

    CAS  Google Scholar 

  • Winstanley, C. and Fothergill, J.L. 2009. The role of quorum sensing in chronic cystic fibrosis Pseudomonas aeruginosa infections. FEMS Microbiol. Lett. 290, 1–9.

    Article  CAS  PubMed  Google Scholar 

  • Yonezawa, A., Kuwahara, J., Fujii, N., and Sugiura, Y. 1992. Binding of tachyplesin I to DNA revealed by footprinting analysis: significant contribution of secondary structure to DNA binding and implication for biological action. Biochemistry 31, 2998–3004.

    Article  CAS  PubMed  Google Scholar 

  • Zhao, T. and Liu, Y. 2010. N-acetylcysteine inhibit biofilms produced by Pseudomonas aeruginosa. BMC Microbiol. 10, 140.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zulianello, L., Canard, C., Köhler, T., Caille, D., Lacroix, J.S., and Meda, P. 2006. Rhamnolipids are virulence factors that promote early infiltration of primary human airway epithelia by Pseudomonas aeruginosa. Infect. Immun. 74, 3134–3147.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Dr. Amal Emad (Microbiology Department, Faculty of Pharmacy, Future University) which supported us with the standard strain P. aeruginosa PAO1.

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, Faculty of Pharmacy, Nahda University, Beni-Suef, Egypt

    Mostafa N. Taha

  2. Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Menoufia University, Menoufia, Egypt

    Amal E. Saafan

  3. Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt

    A. Ahmedy

  4. Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt

    Eman El Gebaly & Ahmed S. Khairalla

Authors
  1. Mostafa N. Taha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amal E. Saafan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Ahmedy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Eman El Gebaly
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ahmed S. Khairalla
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mostafa N. Taha.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Taha, M.N., Saafan, A.E., Ahmedy, A. et al. Two novel synthetic peptides inhibit quorum sensing-dependent biofilm formation and some virulence factors in Pseudomonas aeruginosa PAO1. J Microbiol. 57, 618–625 (2019). https://doi.org/10.1007/s12275-019-8548-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12275-019-8548-2

Keywords

Search

Navigation