Evaluation of a new high-throughput method for identifying quorum quenching bacteria

Tang, Kaihao; Zhang, Yunhui; Yu, Min; Shi, Xiaochong; Coenye, Tom; Bossier, Peter; Zhang, Xiaohua

doi:10.1038/srep02935

Cited by 71 publications

(64 citation statements)

References 56 publications

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“…In a previous study, we demonstrated that the flounder mucusderived strain Muricauda olearia Th120 has strong AHL degradative activity (14) and could decrease biofilm formation and virulence in P. aeruginosa PAO1 (19). In the present study, we identified the momL gene that encodes a novel secretory AHL lactonase from Th120.…”

mentioning

confidence: 67%

“…The cell suspension was sonicated on ice, and the supernatant and cell content were collected after centrifugation and filtration. The C 6 -and C 12 -HSL degradative activities of these sample were measured using the A136 liquid X-Gal assay, which applies 1,4-piperazinediethanesulfonic acid (PIPES) buffer (pH 6.7) to prevent alkaline hydrolysis (14). In addition, the acidification test was used to demonstrate whether the AHL degradation was due to lactonase activity (6).…”

Section: Methodsmentioning

confidence: 99%

“…Within the Bacteroidetes, seven species of QQ bacteria all belonging to the class Flavobacteriia have been identified (14).…”

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confidence: 99%

“…Most of these QQ bacteria were isolated from the marine environment (14)(15)(16)(17), and only one was derived from soil (the potato root-associated Chryseobacterium sp. strain StRB126) (18).…”

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confidence: 99%

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MomL, a Novel Marine-Derived N -Acyl Homoserine Lactonase from Muricauda olearia

Tang

Brackman

et al. 2015

Appl Environ Microbiol

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bGram-negative bacteria use N-acyl homoserine lactones (AHLs) as quorum sensing (QS) signaling molecules for interspecies communication, and AHL-dependent QS is related with virulence factor production in many bacterial pathogens. Quorum quenching, the enzymatic degradation of the signaling molecule, would attenuate virulence rather than kill the pathogens, and thereby reduce the potential for evolution of drug resistance. In a previous study, we showed that Muricauda olearia Th120, belonging to the class Flavobacteriia, has strong AHL degradative activity. In this study, an AHL lactonase (designated MomL), which could degrade both short-and long-chain AHLs with or without a substitution of oxo-group at the C-3 position, was identified from Th120. Liquid chromatography-mass spectrometry analysis demonstrated that MomL functions as an AHL lactonase catalyzing AHL degradation through lactone hydrolysis. MomL is an AHL lactonase belonging to the metallo-␤-lactamase superfamily that harbors an N-terminal signal peptide. The overall catalytic efficiency of MomL for C 6 -HSL is ϳ2.9 ؋ 10 5 s ؊1 M ؊1 . Metal analysis and site-directed mutagenesis showed that, compared to AiiA, MomL has a different metal-binding capability and requires the histidine and aspartic acid residues for activity, while it shares the "HXHXDH" motif with other AHL lactonases belonging to the metallo-␤-lactamase superfamily. This suggests that MomL is a representative of a novel type of secretory AHL lactonase. Furthermore, MomL significantly attenuated the virulence of Pseudomonas aeruginosa in a Caenorhabditis elegans infection model, which suggests that MomL has the potential to be used as a therapeutic agent. N-Acyl homoserine lactones (AHLs) are quorum-sensing (QS) signaling molecules that are used by many Gram-negative bacteria to communicate within species, to regulate gene expression and to synchronize social behaviors, such as biofilm formation, bioluminescence, and secretion of virulence factors (1, 2). An AHL molecule typically consists of a homoserine lactone and an acyl chain with an even number of carbons, with an occasional modification (hydroxy or olefinic double bond) at the C-3 position (1). It has been well established that AHL-dependent QS regulates virulence factor production in many bacterial pathogens, such as Pseudomonas aeruginosa, Erwinia carotovora, Vibrio spp., and Burkholderia spp. (1). Interference with QS has been recognized as a promising antivirulence therapy. Disturbing the QS systems in these pathogens would attenuate virulence rather than kill the bacteria and thereby weaken the selective pressure imposed on the pathogens and reduce the potential for evolution of drug resistance (3). QS inhibitors (QSIs; small molecules) and quorum-quenching (QQ) enzymes can both be used to interfere with QS. QSIs generally act to inactivate autoinducer synthases or receptors through competitive binding, whereas QQ enzymes switch off signal transmission through degradation of the signaling molecules. It has been demonstrated th...

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confidence: 67%

Section: Methodsmentioning

confidence: 99%

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MomL, a Novel Marine-Derived N -Acyl Homoserine Lactonase from Muricauda olearia

Tang

Brackman

et al. 2015

Appl Environ Microbiol

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116

129

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“…These strains often lack the ability to produce native QS signals; however, they are able to respond to exogenous autoinducers, often with a clearly detectable phenotype, such as violacein pigment production in Chromobacterium violaceum CV026 (85,86) and bioluminescence production in V. harveyi (87) or Agrobacterium tumefaciens A136 (88). Using such reporter strains, numerous compounds degrading or modifying AHL have been identified, including signal molecules and AHL analogues (reviewed in references 16 and 19).…”

Section: Figmentioning

confidence: 99%

Novel Reporter for Identification of Interference with Acyl Homoserine Lactone and Autoinducer-2 Quorum Sensing

Weiland‐Bräuer

Pinnow

Schmitz

2015

Appl Environ Microbiol

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Two reporter strains were established to identify novel biomolecules interfering with bacterial communication (quorum sensing [QS]). The basic design of these Escherichia coli-based systems comprises a gene encoding a lethal protein fused to promoters induced in the presence of QS signal molecules. Consequently, these E. coli strains are unable to grow in the presence of the respective QS signal molecules unless a nontoxic QS-interfering compound is present. The first reporter strain designed to detect autoinducer-2 (AI-2)-interfering activities (AI2-QQ.1) contained the E. coli ccdB lethal gene under the control of the E. coli lsrA promoter. The second reporter strain (AI1-QQ.1) contained the Vibrio fischeri luxI promoter fused to the ccdB gene to detect interference with acyl-homoserine lactones. Bacteria isolated from the surfaces of several marine eukarya were screened for quorum-quenching (QQ) activities using the established reporter systems AI1-QQ.1 and AI2-QQ.1. Out of 34 isolates, two interfered with acylated homoserine lactone (AHL) signaling, five interfered with AI-2 QS signaling, and 10 were demonstrated to interfere with both signal molecules. Open reading frames (ORFs) conferring QQ activity were identified for three selected isolates (Photobacterium sp., Pseudoalteromonas sp., and Vibrio parahaemolyticus). Evaluation of the respective heterologously expressed and purified QQ proteins confirmed their ability to interfere with the AHL and AI-2 signaling processes. Q uorum sensing (QS) is the cell-cell communication betweenbacteria that allows the perception of population density by small signaling molecules-so-called autoinducers-and modifies gene expression in response to the population density. It controls a wide spectrum of processes and phenotypic behaviors, including stress resistance, production of toxins and secondary metabolites, pathogenicity, swarming, and biofilm formation (for a review, see references 1 and 2). In addition to playing important roles in intraspecies and interspecies communication, QS is also involved in host-microbe interactions (3-5). Intraspecific communication of Gram-negative bacteria is based on the production and perception of acylated homoserine lactones (AHLs) synthesized by LuxI homologs. Increasing intracellular concentrations of diffusible AHLs with higher cell densities are perceived by binding of the AHLs to the cognate receptor (e.g., LuxR in Vibrio fischeri), resulting in activation or inhibition of target gene transcription (6, 7). Interspecific communication between different species within one habitat has been demonstrated to depend on the synthesis and detection of the universal autoinducer-2 (AI-2), which can be inhibited by furanones (8, 9). The perception of these signal molecules is achieved either by a two-component regulatory system (e.g., in Vibrio harveyi, where the induced phosphorylation cascade finally results in the induction of specific target genes) (10), or by transportation of the signal molecule into the cell (e.g., in Escherichia coli,...

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Natural Quorum Sensing Inhibitors – Small Molecules, Big Messages

Delago

Mandabi

Meijler

2015

Israel Journal of Chemistry

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Due to the increasing emergence of antibiotic resistant bacterial strains within the past few decades, bacterial infections in general – and hospital‐borne infections, in particular – have become increasingly difficult to fight. It is therefore crucial to find new strategies to fight pathogenic bacteria. Targeted inhibition of quorum sensing (QS) presents a promising alternative. QS is a cell density‐dependent signaling pathway used for intra‐ and interspecies coordination of gene expression. In many bacteria, pathogenic phenotypes, as well as the expression of virulence factors, are under the control of QS regulons. A closer look at natural quorum sensing inhibitors may be helpful to identify potent compounds that can be used as alternatives to antibiotics. Moreover, it will also provide insight into the interactions between species that compete for the same habitat and resources. This review aims to summarize our current knowledge concerning natural QS inhibitors, as a starting point for the design and synthesis of new therapeutics to treat or prevent bacterial infections.

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Evaluation of a new high-throughput method for identifying quorum quenching bacteria

Cited by 71 publications

References 56 publications

MomL, a Novel Marine-Derived N -Acyl Homoserine Lactonase from Muricauda olearia

MomL, a Novel Marine-Derived N -Acyl Homoserine Lactonase from Muricauda olearia

Novel Reporter for Identification of Interference with Acyl Homoserine Lactone and Autoinducer-2 Quorum Sensing

Natural Quorum Sensing Inhibitors – Small Molecules, Big Messages

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