Metagenomics pp 271-286

Part of the Methods in Molecular Biology book series (MIMB, volume 1539) | Cite as

Screening for N-AHSL-Based-Signaling Interfering Enzymes

Protocol

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.

Key words

N-acyl homoserine lactone Quorum sensing Quorum quenching N-AHSL lactonase N-AHSL acylase N-AHSL amidohydrolase 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Interactions Arbres MicroorganismesINRA Université de LorraineChampenouxFrance
  2. 2.Univ LyonINSA-Lyon, UCBLVilleurbanne CedexFrance
  3. 3.Univ Lyon, ENS-LyonCNRS UMR5276LyonFrance

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