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Microbiology 148 (2002), 87-102
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Microbiology (2002), 148, 87-102.
© 2002 Society for General Microbiology

Research Paper

Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound

Morten Hentzer1, Kathrin Riedel2, Thomas B. Rasmussen1, Arne Heydorn1, Jens Bo Andersen1, Matthew R. Parsek3, Scott A. Rice4, Leo Eberl2, Søren Molin1, Niels Høiby5, Staffan Kjelleberg4 and Michael Givskov1

Department of Molecular Microbiology, BioCentrum-DTU, Technical University of Denmark, DK-2800 Lyngby, Denmark1
Lehrstuhl für Mikrobiologie, Technische Universität München, D-85350 Freising, Germany2
Department of Civil Engineering, Technological Institute, North Western University, Evanston, IL 60208-3109, USA3
School of Microbiology and Immunology, University of New South Wales (UNSW), Australia 20524
Department of Clinical Microbiology, University Hospital (Rigshospitalet), DK-2100 Copenhagen, Denmark5

Author for correspondence: Michael Givskov. Tel: +45 4525 2768. Fax: +45 4593 2809. e-mail: michael.givskov{at}biocentrum.dtu.dk

Novel molecular tools have been constructed which allow for in situ detection of N-acyl homoserine lactone (AHL)-mediated quorum sensing in Pseudomonas aeruginosa biofilms. The reporter responds to AHL activation of LasR by expression of an unstable version of the green-fluorescent protein (Gfp). Gfp-based reporter technology has been applied for non-destructive, single-cell-level detection of quorum sensing in laboratory-based P. aeruginosa biofilms. It is reported that a synthetic halogenated furanone compound, which is a derivative of the secondary metabolites produced by the Australian macroalga Delisea pulchra, is capable of interfering with AHL-mediated quorum sensing in P. aeruginosa. It is demonstrated that the furanone compound specifically represses expression of a PlasB-gfp reporter fusion without affecting growth or protein synthesis. In addition, it reduces the production of important virulence factors, indicating a general effect on target genes of the las quorum sensing circuit. The furanone was applied to P. aeruginosa biofilms established in biofilm flow chambers. The Gfp-based analysis reveals that the compound penetrates microcolonies and blocks cell signalling and quorum sensing in most biofilm cells. The compound did not affect initial attachment to the abiotic substratum. It does, however, affect the architecture of the biofilm and enhances the process of bacterial detachment, leading to a loss of bacterial biomass from the substratum.

Keywords: biofilms, quorum sensing inhibitors, green-fluorescent protein

Abbreviations: AHL, N-acyl-homoserine lactone; BHL, N-butanoyl-L-homoserine lactone; Gfp, green-fluorescent protein; OdDHL, N-[3-oxo-dodecanoyl]-L-homoserine lactone; OHHL, N-[3-oxo-hexanoyl]-L-homoserine lactone; QSC, quorum-sensing-controlled; QSI, quorum sensing inhibitor; Rfp, red-fluorescent protein; SCLM, scanning confocal laser microscopy




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