Nanoscale visualisation of bacteriocins in action on bacterial membranes

细菌素在细菌膜上作用的纳米级可视化

• 批准号: 2870122
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2870122
• 关键词: Nanoscale; visualisation; bacteriocins; action; bacterial

The rise in antimicrobial-resistant "superbugs" urges us to expand the repertoire of antimicrobial strategies. Of particular interest are antibiotics that target bacterial membranes, since resistance to these antibiotics is hard to develop without compromising bacterial viability overall. Bacteriocins are promising candidates: e.g. nisin is already widely used as a food preservative that targets Gram-positive bacteria, but is not (yet) effective against Gram-negative bacteria, and e.g. epidermicin shows wide tuneability in its modes of membrane disruption. However, mechanisms of action on real bacterial membranes remain poorly understood, and substantial bioengineering remains necessary to reach the specificity and broad-range effectiveness required for next-generation antibiotics.This project is based on the presumption that such bioengineering and prospective wider application will be facilitated by a nanoscale understanding of mechanisms of action, and on the hypothesis that this understanding can be achieved by an integrative microscopy approach applied to living bacteria.Our objectives are:To prepare functional, fluorescently labelled bacteriocin constructs (based on nisin, epidermicin) and to visualise their modes of action by AFM on supported lipid bilayers.To develop and apply experimental protocols to image binding bacteriocins on model membranes by single-molecule total internal reflection fluorescence (TIRF) microscopy.To visualise these bacteriocins and their modes of action by AFM on live bacteria.To develop and apply experimental protocols to image bacteriocins on live bacteria, as well as the membrane disruption (using lipid and nucleic acid staining) by fluorescence microscopy, as well as stop-flow experiments for super-resolution experiments (PALM/STORM).To use these methods to correlate molecular design with modes of membrane disruption and bacterial killing.

抗菌抗菌的“超级细菌”的增长敦促我们扩大抗菌策略的曲目。特别令人感兴趣的是靶向细菌膜的抗生素，因为对这些抗生素的耐药性很难在不损害细菌生存力的情况下发展。细菌素是有希望的候选人：例如Nisin已被广泛用作靶向革兰氏阳性细菌的食品防腐剂，但尚未（尚未）对革兰氏阴性细菌有效，例如表皮素在其膜破坏模式下显示出广泛的可调性。然而，对真实细菌膜的作用机制仍然很众所周知，并且要达到下一代抗生素所需的特异性和宽距离的实质性工程仍然是必要的。该项目基于这样的假定，通过对纳米级的机制的理解，对这种能力的理解将促进这种能力，并在纳米级的范围内促进了这种方法，并且可以通过纳米级的机制来促进，并促进了这种方法，并促进了纳米级的作用。应用于活细菌。我们的目标是：准备功能性的，荧光标记的细菌蛋白构建体（基于Nisin，表皮蛋白），并可视化AFM对AFM的作用方式，以开发和应用实验方案来开发和应用实验方案，以对这些反射型细菌图像型型细菌（模型膜）在模型中进行图像，以使模型（模型膜）在模型中进行构图（细菌素及其对活细菌的作用模式。要开发和应用实验方案，以通过荧光显微镜进行膜的细菌以及膜破坏（使用脂质和核酸染色）来形象，并通过荧光显微镜进行膜破坏（使用脂质和核酸染色），并与这些模式的超级分辨率实验（对这些方法进行摩擦）。破坏和细菌杀死。