On-line high-performance liquid chromatography–mass spectrometric detection and quantification of N-acylhomoserine lactones, quorum sensing signal molecules, in the presence of biological matrices

Morin, Danièle; Grasland, Béatrice; Vallée‐Réhel, Karine; Dufau, Chrystèle; Haras, Dominique

doi:10.1016/s0021-9673(03)00730-1

Cited by 163 publications

(162 citation statements)

References 20 publications

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“…The mobile phase consisted of a linear gradient (40-95%) of solvent B (methanol with 0.1% formic acid) and solvent A (25 mM ammonium formate with 0.1% formic acid). Effluents were ionized by electrospray ionization under positive mode and detected using the multiple reaction monitoring approach (Morin et al, 2003;Decho et al, 2009). Matrix-matched multiple reaction monitoring experiments were conducted for all 13 standard AHLs (Supplementary Table S1).…”

Section: Chemicalsmentioning

confidence: 99%

The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules

et al. 2014

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Quorum sensing (QS) signalling has been extensively studied in single species populations. However, the ecological role of QS in complex, multi-species communities, particularly in the context of community assembly, has neither been experimentally explored nor theoretically addressed. Here, we performed a long-term bioreactor ecology study to address the links between QS, organization and composition of complex microbial communities. The conversion of floccular biomass to highly structured granules was found to be non-random, but strongly and positively correlated with N-acyl-homoserine-lactone (AHL)-mediated QS. Specific AHLs were elevated up to 100-fold and were strongly associated with the initiation of granulation. Similarly, the levels of particular AHLs decreased markedly during the granular disintegration phase. Metadata analysis indicated that granulation was accompanied by changes in extracellular polymeric substance (EPS) production and AHL add-back studies also resulted in increased EPS synthesis. In contrast to the commonly reported nanomolar to micromolar signal concentrations in pure culture laboratory systems, QS signalling in the granulation ecosystem occurred at picomolar to nanomolar concentrations of AHLs. Given that low concentrations of AHLs quantified in this study were sufficient to activate AHL bioreporters in situ in complex granular communities, AHL mediated QS may be a common feature in many natural and engineered ecosystems, where it coordinates community behaviour.

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Section: Chemicalsmentioning

confidence: 99%

The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules

et al. 2014

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“…Linear-quadrupole ion trap devices are capable of secondary mass spectrometry, enabling fine structural investigations of unknown analytes (e.g. Morin et al, 2003;Ortori et al, 2007). Finally, triple quadrupole devices are extremely sensitive and selective; they are capable of operating in a selected reaction monitoring (SRM) mode which allows quantitative observation of specific ions amidst an extremely complex matrix.…”

Section: Analytical Approaches To the Investigation Of Qs In Natural mentioning

confidence: 99%

Microbial interactions associated with biofilms attached to Trichodesmium spp. and detrital particles in the ocean

Hmelo¹

2010

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Quorum sensing (QS) via acylated homoserine lactones (AHLs) was discovered in the ocean, yet little is known about its role in the ocean beyond its involvement in certain symbiotic interactions. The objectives of this thesis were to constrain the chemical stability of AHLs in seawater, explore the production of AHLs in marine particulate environments, and probe selected behaviors which might be controlled by AHL-QS. I established that AHLs are more stable in seawater than previously expected and are likely to accumulate within biofilms. Based on this result, I chose to study AHL-QS in the bacterial communities inhabiting biofilms attached to Trichodesmium spp. and detrital (photosynthetically-derived sinking particulate organic carbon, POC) particles. These hot spots of microbial activity are primary sites of interaction between marine primary producers and heterotrophs and crucial components of the biological pump.Biofilm communities associated with Trichodesmium thiebautii colonies in the Sargasso Sea differed considerably from seawater microbial communities. In addition, there was no overlap between the communities associated with tuft and puff colonies. These results suggest that bacterial communities associated with Trichodesmium are not random; rather, Trichodesmium selects for specific microbial flora. Novel 16S rRNA gene sequences are present both in clone libraries constructed from DNA extracted from colonies of Trichodesmium spp. and in culture collections derived from wild and laboratory cultivated Trichodesmium spp., supporting the idea that the phycosphere of Trichodesmium is a unique microenvironment.Using high performance liquid chromatography-mass spectrometry, I demonstrated that bacteria isolated from Trichodesmium synthesize AHLs. In addition, I detected AHLs in sinking particles collected from a site off of Vancouver Island, Canada. AHLs were subsequently added to laboratory cultures of non-axenic Trichodesmium colonies and sinking POC samples. This is the first time AHLs have been detected in POC and indicates that AHL-QS was occurring in POC. Further, I showed that AHLs enhanced certain organic-matter degrading hydrolytic enzyme activities. My results suggest that AHL-QS is a factor regulating biogeochemically relevant enzyme activities on sinking POC and within the biofilms attached to Trichodesmium colonies and thereby may impact the timing and magnitude of biogeochemical fluxes in the ocean.

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“…After removal of Na 2 SO 4 crystals by centrifugation , the solvent was evaporated. The samples were then treated as described previously (Morin et al, 2003) and analysed by C 18 reversed-phase HPLC/MS analysis using an Agilent Technologies 1100 system coupled to a Bruker Esquire-LC ESI-MS/ MS positive-ion electrospray ionization and ion-trap mass spectrometer.…”

Section: Methodsmentioning

confidence: 99%

Gamma-aminobutyric acid acts as a specific virulence regulator in Pseudomonas aeruginosa

et al. 2013

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Gamma-aminobutyric acid (GABA) is widespread in the environment and can be used by animal and plants as a communication molecule. Pseudomonas species, in particular fluorescent ones, synthesize GABA and express GABA-binding proteins. In this study, we investigated the effects of GABA on the virulence of Pseudomonas aeruginosa. While exposure to GABA (10 mM) did not modify either the growth kinetics or the motility of the bacterium, its cytotoxicity and virulence were strongly increased. The Caenorhabditis elegans 'fast killing test' model revealed that GABA acts essentially through an increase in diffusible toxin(s). GABA also modulates the biofilm formation activity and adhesion properties of PAO1. GABA has no effect on cell surface polarity, biosurfactant secretion or on the lipopolysaccharide structure. The production of several exoenzymes, pyoverdin and exotoxin A is not modified by GABA but we observed an increase in cyanogenesis which, by itself, could explain the effect of GABA on P. aeruginosa virulence. This mechanism appears to be regulated by quorum sensing. A proteomic analysis revealed that the effect of GABA on cyanogenesis is correlated with a reduction of oxygen accessibility and an over-expression of oxygen-scavenging proteins. GABA also promotes specific changes in the expression of thermostable and unstable elongation factors Tuf/Ts involved in the interaction of the bacterium with the host proteins. Taken together, these results suggest that GABA is a physiological regulator of P. aeruginosa virulence.

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On-line high-performance liquid chromatography–mass spectrometric detection and quantification of N-acylhomoserine lactones, quorum sensing signal molecules, in the presence of biological matrices

Cited by 163 publications

References 20 publications

The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules

The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules

Microbial interactions associated with biofilms attached to Trichodesmium spp. and detrital particles in the ocean

Gamma-aminobutyric acid acts as a specific virulence regulator in Pseudomonas aeruginosa

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