Kobras Carolin Martina, Mascher Thorsten, Gebhard Susanne
Department of Biology and Biochemistry, Milner Centre for Evolution, University of Bath, Claverton Down, Bath, BA2 7AY, UK.
Institut für Mikrobiologie, Technische Universität Dresden, 01062, Dresden, Germany.
Methods Mol Biol. 2017;1520:121-131. doi: 10.1007/978-1-4939-6634-9_7.
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 , 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.
基于报告菌株对特定刺激的反应可视化的全细胞生物传感器，是监测特定环境条件或化合物存在的一种强大且经济高效的手段。此类生物传感器经常应用的一个特定领域是药物发现，即筛选大量细菌或真菌菌株以生产抗菌化合物。我们在此描述一种基于发光的枯草芽孢杆菌生物传感器用于发现细胞壁活性物质的应用。该系统基于特征明确的启动子P，它在响应导致细胞包膜应激的多种条件时被诱导，特别是干扰细胞壁生物合成膜锚定步骤的抗生素。一种简单的“点种法”检测，即将潜在生产菌株的菌落在报告菌株的菌苔上直接培养，能够对抗菌化合物进行定量和时间分辨检测。由于该方法对技术要求极低，我们预计它很容易应用于多种候选生产菌株和生长条件。
