Abstract
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF), a non-halogenated furanone found in a variety of fruits, has been shown to have antimicrobial activity. However, few studies have focused on its inhibitory effect on bacterial quorum sensing (QS) at levels below the non-inhibitory concentration. In this study, 0.1 μM HDMF decreased the production of QS signal molecules and inhibited QS-controlled biofilm formation by Pseudomonas aeruginosa PAO1 without causing growth inhibition. In the presence of 0.1 and 1.0 μM HDMF, biofilm production by PAO1 was reduced by 27.8 and 42.6%, respectively, compared to that by untreated control cells. HDMF (1.0 μM) also significantly affected virulence factor expression (regulated by the las, rhl, and pqs system), resulting in a significant reduction in the production of LasA protease (53.8%), rhamnolipid (40.9%), and pyocyanin (51.4%). This HDMF-dependent inhibition of virulence factor expression was overcome by increasing the levels of two QS signal molecules of P. aeruginosa, N-(3-oxo-dodecanoyl)-L-homoserine lactone and N-butyryl-L-homoserine lactone, suggesting reversible competitive inhibition between HDMF and these molecules. The results of this study indicate that HDMF has great potential as an inhibitor of QS, and that it may be of value as a therapeutic agent and in biofilm control, without increasing selective pressure for resistance development.
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Adonizio, A., Kong, K.-F., and Mathee, K. 2008. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by South Florida plant extracts. Antimicrob. Agents Chemother. 52, 198–203.
Caiazza, N.C., Shanks, R.M., and O’Toole, G.A. 2005. Rhamnolipids modulate swarming motility patterns of Pseudomonas aeruginosa. J. Bacteriol. 187, 7351–7361.
Daniels, R., Vanderleyden, J., and Michiels, J. 2004. Quorum sensing and swarming migration in bacteria. FEMS Microbiol. Rev. 28, 261–289.
Davies, D.G., Parsek, M.R., Pearson, J.P., Iglewski, B.H., Costerton, J.W., and Greenberg, E.P. 1998. The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280, 295–298.
Dong, Y.H., Wang, L.H., Xu, J.L., Zhang, H.B., Zhang, X.F., and Zhang, L.H. 2001. Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase. Nature 411, 813–817.
Doyle, T.B., Hawkins, A.C., and McCarter, L.L. 2004. The complex flagella torque generator of Pseudomonas aeruginosa. J. Bacteriol. 186, 6341–6350.
Dubin, P.J. and Kolls, J.K. 2007. Pseudomonas aeruginosa and the host pulmonary immune response. Expert Rev. Respir. Med. 1, 121–137.
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.
Gule, N.P., De Kwaadsteniet, M., Cloete, T.E., and Klumperman, B. 2013. Furanone-containing poly(vinyl alcohol) nanofibers for cell-adhesion inhibition. Wat. Res. 47, 1049–1059.
Hentzer, M. and Givskov, M. 2003. Pharmacological inhibition of quorum sensing for the treatment of chronic bacterial infections. J. Clin. Invest. 112, 1300–1307.
Hentzer, M., Riedel, K., Rasmussen, T.B., Heydorn, A., Andersen, J.B., Parsek, M.R., Rice, S.A., Eberl, L., Molin, S., HØiby, N., and et al. 2002. Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound. Microbiology 148, 87–102.
Hentzer, M., Wu, H., Andersen, J.B., Riedel, K., Rasmussen, T.B., Bagge, N., Kumar, N., Schembri, M.A., Song, Z., Kristoffersen, P., and et al. 2003. Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO, J. 22, 3803–3815.
Hoffman, L.R., D’Argenio, D.A., MacCoss, M.J., Zhang, Z., Jones, R.A., and Miller, S.I. 2005. Aminoglycoside antibiotics induce bacterial biofilm formation. Nature 436, 1171–1175.
Janssens, J.C.A., Steenackers, H., Robijns, S., Gellens, E., Levin, J., Zhao, H., Hermans, K., De Coster, D., Verhoeven, T.L., Marchal, K., and et al. 2008. Brominated furanones inhibit biofilm formation by Salmonella enterica serovar Typhimurium. Appl. Environ. Microbiol. 74, 6639–6648.
Khan, M.S.A., Zahin, M., Hasan, S., Husain, F.M., and Ahmad, I. 2009. Inhibition of quorum sensing regulated bacterial functions by plant essential oils with special reference to clove oil. Lett. Appl. Microbiol. 49, 354–360.
Kim, C., Kim, J., Park, H.Y., Park, H.J., Lee, J.H., Kim, C.K., and Yoon, J. 2008. Furanone derivatives as quorum-sensing antagonists of Pseudomonas aeruginosa. Appl. Microbiol. Biotechnol. 80, 37–47.
Köhler, T., Curty, L.K., Barja, F., van Delden, C., and Pechère, J.C. 2000. Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J. Bacteriol. 182, 5990–5996.
Kong, K.F., Jayawardena, S.R., Indulkar, S.D., del Puerto, A., Koh, C.L., HØiby, N., and Mathee, K. 2005. Pseudomonas aeruginosa AmpR is a global transcriptional factor that regulates expression of AmpC and PoxB β-lactamases, proteases, quorum sensing, and other virulence factors. Antimicrob. Agents Chemother. 49, 4567–4575.
Luo, Z.Q., Su, S., and Farrand, S.K. 2003. In situ activation of the quorum-sensing transcription factor TraR by cognate and noncognate acy l-homoserine lactone ligands: kinetics and consequences. J. Bacteriol. 185, 5665–5672.
Manefield, M., de Nys, R., Kumar, N., Read, R., Givskov, M., Steinberg, P., and Kjelleberg, S. 1999. Evidence that halogenated furanones from Delisea pulchra inhibit acylated homoserine lactone (AHL)-mediated gene expression by displacing the AHL signal from its receptor protein. Microbiology 145, 283–291.
Manefield, M., Rasmussen, T.B., Hentzer, M., Andersen, J.B., Steinberg, P., Kjelleberg, S., and Givskov, M. 2002. Halogenated furanones inhibit quorum sensing through accelerated LuxR turnover. Microbiology 148, 1119–1127.
Manefield, M., Welch, M., Givskov, M., Salmond, G.P.C., and Kjelleberg, S. 2001. Halogenated furanones from the red alga, Delisea pulchra, inhibit carbapenem antibiotic synthesis and exoenzyme virulence factor production in the phytopathogen Erwinia carotovora. FEMS Microbiol. Lett. 205, 131–138.
Martinelli, D., Grossmann, G., Séquin, U., Brandl, H., and Bachofen, R. 2004. Effects of natural and chemically synthesized furanones on quorum sensing in Chromobacterium violaceum. BMC Microbiol. 4, 25–34.
McClean, K.H., Winson, M.K., Fish, L., Taylor, A., Chhabra, S.R., Camara, M., Daykin, M., Lamb, J.H., Swift, S., Bycroft, B.W., and et al. 1997. Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143, 3703–3711.
de Nys, R., Wright, A.D., König, G.M., and Sticher, O. 1993. New halogenated furanones from the marine alga Delisea pulchra (cf. fimbriata). Tetrahedron 49, 11213–11220.
O’Toole, G.A. and Kolter, R. 1998. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol. Microbiol. 28, 449–461.
Overhage, J., Lewenza, S., Marr, A.K., and Hancock, R.E.W. 2007. Identification of genes involved in swarming motility using a Pseudomonas aeruginosa PAO1 mini-Tn5-lux mutant library. J. Bacteriol. 189, 2164–2169.
Parsek, M.R., Val, D.L., Hanzelka, B.L., Cronan, J.E. Jr., and Greenberg, E.P. 1999. Acyl homoserine-lactone quorum-sensing signal generation. Proc. Natl. Acad. Sci. USA 96, 4360–4365.
Paul, D., Kim, Y.S., Ponnusamy, K., and Kweon, J.H. 2009. Application of quorum quenching to inhibit biofilm formation. Environ. Eng. Sci. 26, 1319–1324.
Pearson, J.P., Pesci, E.C., and 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.
Ponnusamy, K., Paul, D., Kim, Y.S., and Kweon, J.H. 2010. 2(5H)-Furanone: a prospective strategy for biofouling-control in membrane biofilm bacteria by quorum sensing inhibition. Braz. J. Microbiol. 41, 227–234.
Rasch, M., Buch, C., Austin, B., Slierendrecht, W.J., Ekmann, K.S., Larsen, J.L., Johansen, C., Riedel, K., Eberl, L., Givskov, M., and et al. 2004. An inhibitor of bacterial quorum sensing reduces mortalities caused by vibriosis in rainbow trout (Oncorhyncus mykiss, Walbaum). Syst. Appl. Microbiol. 27, 350–359.
Rasmussen, T.B., Bjarnsholt, T., Skindersoe, M.E., Hentzer, M., Kristoffersen, P., Köte, M., Nielsen, J., Eberl, L., and Givskov, M. 2005. Screening for quorum-sensing inhibitors (QSI) by use of a novel genetic system, the QSI selector. J. Bacteriol. 187, 1799–1814.
Rasmussen, T.B., Manefield, M., Andersen, J.B., Eberl, L., Anthoni, U., Christophersen, C., Steinberg, P., Kjelleberg, S., and Givskov, M. 2000. How Delisea pulchra furanones affect quorum sensing and swarming motility in Serratia liquefaciens MG1. Microbiology 146, 3237–3244.
Ravn, L., Christensen, A.B., Molin, S., Givskov, M., and Gram, L. 2001. Methods for detecting acylated homoserine lactones produced by Gram-negative bacteria and their application in studies of AHL-production kinetics. J. Microbiol. Methods 44, 239–251.
Reimmann, C., Ginet, N., Michel, L., Keel, C., Michaux, P., Krishnapillai, V., Zala, M., Heurlier, K., Triandafillu, K., Harms, H., and et al. 2002. Genetically programmed autoinducer destruction reduces virulence gene expression and swarming motility in Pseudomonas aeruginosa PAO1. Microbiology 148, 923–932.
Sasaki, T., Yamakoshi, J., Saito, M., Kasai, K., Matsudo, T., Koga, T., and Mori, K. 1998. Antioxidative activities of 4-hydroxy-3(2H)-furanones and their anticataract effect on spontaneous cataract rat (ICR/f). Biosci. Biotechnol. Biochem. 62, 1865–1869.
Schaadt, N.S., Steinbach, A., Hartmann, R.W., and Helms, V. 2013. Rule-based regulatory and metabolic model for quorum sensing in P. aeruginosa. BMC Syst. Biol. 7, 81–94.
Shetye, G.S., Singh, N., Gao, X., Bandyopadhyay, D., Yan, A., and Luk, Y.Y. 2013. Structures and biofilm inhibition activities of brominated furanones for Escherichia coli and Pseudomonas aeruginosa. Med. Chem. Commun. 4, 1079–1084.
Shobharani, P. and Agrawal, R. 2010. Interception of quorum sensing signal molecule by furanone to enhance shelf life of fermented milk. Food Control 21, 61–69.
Shrout, J.D., Chopp, D.L., Just, C.L., Hentzer, M., Givskov, M., and Parsek, M.R. 2006. The impact of quorum sensing and swarming motility on Pseudomonas aeruginosa biofilm formation is nutritionally conditional. Mol. Microbiol. 62, 1264–1277.
Sio, C.F., Otten, L.G., Cool, R.H., Diggle, S.P., Braun, P.G., Bos, R., Daykin, M., Cámara, M., Williams, P., and Quax, W.J. 2006. Quorum quenching by an N-acyl-homoserine lactone acylase from Pseudomonas aeruginosa PAO1. Infect. Immun. 74, 1673–1682.
Slaughter, J.C. 1999. The naturally occurring furanones: formation and function from pheromone to food. Biol. Rev. 74, 259–276.
Smith, R.S. and Iglewski, B.H. 2003a. P. aeruginosa quorum-sensing systems and virulence. Curr. Opin. Microbiol. 6, 56–60.
Smith, R.S. and Iglewski, B.H. 2003b. Pseudomonas aeruginosa quorum sensing as a potential antimicrobial target. J. Clin. Invest. 112, 1460–1465.
Steindler, L. and Venturi, V. 2007. Detection of quorum-sensing Nacyl homoserine lactone signal molecules bybacterial biosensors. FEMS Microbiol. Lett. 266, 1–9.
Stewart, P.S. and Costerton, J.W. 2001. Antibiotic resistance of bacteria in biofilms. Lancet 358, 135–138.
Sung, W.S., Jung, H.J., Park, K., Kim, H.S., Lee, I.S., and Lee, D.G. 2007. 2,5-Dimethyl-4-hydroxy-3(2H)-furanone (DMHF); antimicrobial compound with cell cycle arrest in nosocomial pathogens. Life Sci. 80, 586–591.
Teplitski, M., Robinson, J.B., and Bauer, W.D. 2000. Plants secrete substances that mimic bacterial N-acyl homoserine lactone signal activities and affect population density-dependent behaviors in associated bacteria. Mol. Plant-Microbe Interact. 13, 637–648.
Vandeputte, O.M., Kiendrebeogo, M., Rajaonson, S., Diallo, B., Mol, A., El Jaziri, M., and Baucher, M. 2010. Identification of Catechin as one of the flavonoids from Combretum albiflorum bark extract that reduces the production of quorum-sensingcontrolled virulence factors in Pseudomonas aeruginosa PAO1. Appl. Environ. Microbiol. 76, 243–253.
Whitehead, N.A., Welch, M., and Salmond, G.P.C. 2001. Silencing the majority. Nature Biotechnol. 19, 735–736.
Zhao, Z.G., Yan, S.S., Yu, Y.M., Mi, N., Zhang, L.X., Liu, J., Li, X.L., Liu, F., Xu, J.F., Yang, W.Q., and et al. 2013. An aqueous extract of Yunnan Baiyao inhibits the quorum-sensing-related virulence of Pseudomonas aeruginosa. J. Microbiol. 51, 207–212.
Zhou, L., Zheng, H., Tang, Y., Yu, W., and Gong, Q. 2013. Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnol. Lett. 35, 631–637.
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Choi, SC., Zhang, C., Moon, S. et al. Inhibitory effects of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) on acyl-homoserine lactone-mediated virulence factor production and biofilm formation in Pseudomonas aeruginosa PAO1. J Microbiol. 52, 734–742 (2014). https://doi.org/10.1007/s12275-014-4060-x
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DOI: https://doi.org/10.1007/s12275-014-4060-x