title: Screening for N-AHSL-Based-Signaling Interfering Enzymes.
abstract: Quorum sensing (QS)-based signaling is a widespread pathway used by bacteria for the regulation of functions involved in their relation to the environment or their host. QS relies upon the production, accumulation and perception of small diffusable molecules by the bacterial population, hence linking high gene expression with high cell population densities. Among the different QS signal molecules, an important class of signal molecules is the N-acyl homoserine lactone (N-AHSL). In pathogens such as Erwinia or Pseudomonas, N-AHSL based QS is crucial to overcome the host defenses and ensure a successful infection. Interfering with QS-regulation allows the algae Delisea pulcra to avoid surface colonization by bacteria. Thus, interfering the QS-regulation of pathogenic bacteria is a promising antibiotic-free antibacterial therapeutic strategy. To date, two N-AHSL lactonases and one amidohydrolase families of N-ASHL degradation enzymes have been characterized and have proven to be efficient in vitro to control N-AHSL-based QS-regulated functions in pathogens. In this chapter, we provide methods to screen individual clones or bacterial strains as well as pool of clones for genomic and metagenomic libraries, that can be used to identify strains or clones carrying N-ASHL degradation enzymes.
authors.affiliation: Interactions Arbres Microorganismes, INRA Université de Lorraine, UMR1136, Champenoux, 54280, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Uroz
authors.affiliation: Univ Lyon, INSA-Lyon, UCBL, CNRS UMR5240, 69621, Villeurbanne Cedex, France. poger@ens-lyon.fr.|Univ Lyon, ENS-Lyon, CNRS UMR5276, Lyon, 69634, France. poger@ens-lyon.fr.
authors.forename: Phil M
authors.initials: PM
authors.lastname: Oger
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D013058:Mass Spectrometry; D053038:Quorum Sensing / Q000235:genetics*; D015398:Signal Transduction; D013379:Substrate Specificity
chemical_list: C088386:homoserine lactone; D015107:4-Butyrolactone
keywords: N-AHSL acylase; N-AHSL amidohydrolase; N-AHSL lactonase; N-acyl homoserine lactone; Quorum quenching; Quorum sensing