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Antibiotics pp 121-131 | Cite as

Application of a Bacillus subtilis Whole-Cell Biosensor (PliaI-lux) for the Identification of Cell Wall Active Antibacterial Compounds

  • Carolin Martina Kobras
  • Thorsten Mascher
  • Susanne Gebhard
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1520)

Abstract

Whole-cell biosensors, based on the visualization of a reporter strain’s response to a particular stimulus, are a robust and cost-effective means to monitor defined environmental conditions or the presence of chemical compounds. One specific field in which such biosensors are frequently applied is drug discovery, i.e., the screening of large numbers of bacterial or fungal strains for the production of antimicrobial compounds. We here describe the application of a luminescence-based Bacillus subtilis biosensor for the discovery of cell wall active substances. The system is based on the well-characterized promoter P liaI , which is induced in response to a wide range of conditions that cause cell envelope stress, particularly antibiotics that interfere with the membrane-anchored steps of cell wall biosynthesis. A simple “spot-on-lawn” assay, where colonies of potential producer strains are grown directly on a lawn of the reporter strain, allows for quantitative and time-resolved detection of antimicrobial compounds. Due to the very low technical demands of this procedure, we expect it to be easily applicable to a large variety of candidate producer strains and growth conditions.

Key words

Bio-assay Reporter gene Cell envelope stress Cell wall Antibiotic Antimicrobial peptide Stress response Luminescence Lipid II cycle 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Carolin Martina Kobras
    • 1
  • Thorsten Mascher
    • 2
  • Susanne Gebhard
    • 1
  1. 1.Department of Biology and Biochemistry, Milner Centre for EvolutionUniversity of BathClaverton Down, BathUK
  2. 2.Institut für MikrobiologieTechnische Universität DresdenDresdenGermany

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