Instituto de Agrobiotecnología (IDAB). CSIC- Gobierno de Navarra, Mutilva, Spain.
The Campus 4 Crinan Street London N1, London, UK.
NPJ Biofilms Microbiomes. 2022 Aug 1;8(1):62. doi: 10.1038/s41522-022-00324-w.
Biofilm engineering has emerged as a controllable way to fabricate living structures with programmable functionalities. The amyloidogenic proteins comprising the biofilms can be engineered to create self-assembling extracellular functionalized surfaces. In this regard, facultative amyloids, which play a dual role in biofilm formation by acting as adhesins in their native conformation and as matrix scaffolds when they polymerize into amyloid-like fibrillar structures, are interesting candidates. Here, we report the use of the facultative amyloid-like Bap protein of Staphylococcus aureus as a tool to decorate the extracellular biofilm matrix or the bacterial cell surface with a battery of functional domains or proteins. We demonstrate that the localization of the functional tags can be change by simply modulating the pH of the medium. Using Bap features, we build a tool for trapping and covalent immobilizing molecules at bacterial cell surface or at the biofilm matrix based on the SpyTag/SpyCatcher system. Finally, we show that the cell wall of several Gram-positive bacteria could be functionalized through the external addition of the recombinant engineered Bap-amyloid domain. Overall, this work shows a simple and modulable system for biofilm functionalization based on the facultative protein Bap.
生物膜工程已成为一种可控制的方法，可以制造具有可编程功能的活体结构。生物膜中包含的淀粉样蛋白可以被工程化，以创建自组装的细胞外功能化表面。在这方面，兼性淀粉样蛋白是有趣的候选物，它们在天然构象中作为黏附素在生物膜形成中起作用，而在聚合为类似淀粉样纤维结构时则作为基质支架起作用。在这里，我们报告了使用金黄色葡萄球菌的兼性淀粉样样 Bap 蛋白作为工具，用一系列功能域或蛋白质来修饰细胞外生物膜基质或细菌表面。我们证明，通过简单地调节培养基的 pH 值，可以改变功能标记的定位。使用 Bap 特性，我们构建了一种基于 SpyTag/SpyCatcher 系统的工具，用于在细菌细胞表面或生物膜基质上捕获和共价固定分子。最后，我们表明，通过外部添加重组工程化的 Bap-淀粉样蛋白结构域，可以对几种革兰氏阳性菌的细胞壁进行功能化。总的来说，这项工作展示了一种基于兼性蛋白 Bap 的简单可调节的生物膜功能化系统。
