Application of a Bacillus subtilis Whole-Cell Biosensor (PliaI-lux) for the Identification of Cell Wall Active Antibacterial Compounds.
Application of a Bacillus subtilis Whole-Cell Biosensor (PliaI-lux) for the Identification of Cell Wall Active Antibacterial Compounds.
复制标题
应用枯草芽孢杆菌全细胞生物传感器 (PliaI-lux) 鉴定细胞壁活性抗菌化合物。
DOI：
10.1007/978-1-0716-2855-3_13
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kobras CM
中科院分区：
文献类型：
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作者：
Kobras CM
基于报告菌株对特定刺激的响应的可视化的全细胞生物传感器是监测限定的环境条件或化学化合物的存在的稳健且具有成本效益的手段。这种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用于生产抗微生物化合物。在这里，我们描述了一种基于发光的枯草芽孢杆菌生物传感器在发现细胞壁活性物质方面的应用;这篇文章是对我们2017年发表的上一章的更新。该系统是基于充分表征的启动子PliaI，其响应于引起细胞被膜应激的广泛条件而被诱导，特别是干扰细胞壁生物合成的膜锚定步骤的抗生素。一种简单的“菌苔上点”试验，其中潜在生产菌株的菌落直接生长在报告菌株的菌苔上，允许对抗菌化合物进行定量和时间分辨检测。由于该方法的技术要求非常低，我们预计它很容易适用于各种候选生产菌株和生长条件。
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, that is, the screening of large numbers of bacterial or fungal strains for the production of antimicrobial compounds. Here, we describe the application of a luminescence-basedBacillus subtilisbiosensor for the discovery of cell wall active substances; this article is an update to our previous chapter published in 2017. 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.