title: Application of a Bacillus subtilis Whole-Cell Biosensor (PliaI-lux) for the Identification of Cell Wall Active Antibacterial Compounds.
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 PliaI, 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.
authors.affiliation: Department of Biology and Biochemistry, Milner Centre for Evolution, University of Bath, Claverton Down, Bath, BA2 7AY, UK.
authors.forename: Carolin Martina
authors.initials: CM
authors.lastname: Kobras
authors.affiliation: Institut für Mikrobiologie, Technische Universität Dresden, 01062, Dresden, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Mascher
authors.affiliation: Department of Biology and Biochemistry, Milner Centre for Evolution, University of Bath, Claverton Down, Bath, BA2 7AY, UK. s.gebhard@bath.ac.uk.
authors.forename: Susanne
authors.initials: S
authors.lastname: Gebhard
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000166:cytology* / Q000187:drug effects*; D015374:Biosensing Techniques / Q000379:methods*; D002473:Cell Wall / Q000187:drug effects* / Q000378:metabolism; D017930:Genes, Reporter; D049449:Luminescence; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents
keywords: Antibiotic; Antimicrobial peptide; Bio-assay; Cell envelope stress; Cell wall; Lipid II cycle; Luminescence; Reporter gene; Stress response