PDF(Pubmed)
Abstract：
The use of bacteriocins is a promising approach for addressing the immense threat of food-borne and drug-resistant pathogens. In recent years screening platforms for novel bacteriocins using whole-cell biosensors have been established. During screening cell-to-cell heterogeneity is currently neglected but might play a crucial role in signal development of the whole-cell biosensor after bacteriocin exposure. In this study, we explored the temporal dynamics of the signal heterogeneity of the biosensor Listeria innocua LMG2785/pNZpHin2 Lm after nisin exposure using microfluidic single-cell analysis. The results provided novel and detailed insights into the dynamics of cell-to-cell heterogeneity in L. innocua LMG2785/pNZpHin2 Lm at different nisin concentrations with a high spatio-temporal resolution. Furthermore, the formation of subpopulations during bacteriocin exposure was observed. In-depth single-cell tracking even revealed the regeneration of disrupted cells and recovery of pH homeostasis in rare instances. These findings are highly important for the future design and execution of bacteriocin assays and for the interpretation of fluorescence signal development at the population level after exposure to different concentrations of bacteriocins (here, nisin), as well as for obtaining deeper insights into single-cell persistence strategies to quantify the efficacy and efficiency of novel bacteriocins.
摘要：
使用细菌素是解决食源性和耐药性病原体的巨大威胁的有前途的方法。近年来，已经建立了使用全细胞生物传感器筛选新型细菌素的平台。在筛选过程中，目前忽略了细胞间异质性，但可能在细菌素暴露后全细胞生物传感器的信号发展中起关键作用。在这项研究中,我们使用微流控单细胞分析探索了nisin暴露后生物传感器无毒李斯特菌LMG2785/pNZpHin2Lm的信号异质性的时间动态。结果提供了新的和详细的见解，以不同浓度的乳酸链球菌素LMG2785/pNZpHin2Lm中细胞间异质性的动态，具有高的时空分辨率。此外，观察到细菌素暴露期间亚群的形成。深入的单细胞追踪甚至揭示了在极少数情况下破坏的细胞的再生和pH稳态的恢复。这些发现对于细菌素测定的未来设计和执行以及暴露于不同浓度的细菌素后在人群水平上荧光信号发展的解释非常重要（此处，Nisin),以及更深入地了解单细胞持久性策略，以量化新型细菌素的功效和效率。
