Development and application of a toolbox of biofilm matrix association modules

描述(申请人提供)：我们实验室最近完成了对革兰氏阴性生物膜基质的第一次全面的蛋白质组分析。这项工作证明，生物膜基质中保留了分泌蛋白质的一个子集。对生物膜中起结构作用的三种蛋白质进行了详细的研究。有趣的是，这些蛋白质在生物膜基质中有独特的分布，这与它们在生物膜形成中的功能相对应。更多推测的生物膜相关蛋白的功能和分布仍有待鉴定。 我们假设我们可以利用这些生物膜基质相关蛋白来将功能性酶或抗原靶向生物膜基质的特定区域。在这里，我们建议开发一个生物膜基质结合模块工具箱，并将该工具箱用于原则性实验的证明，在这些实验中，我们将霍乱毒素的几丁质酶分泌酶或B亚单位与生物膜基质在特定的分布中相关联。我们的目标是建立一种新的策略，使蛋白质与革兰氏阴性菌的生物膜基质相结合，用于革兰氏阴性菌的生物医学、生物技术和生物工程应用。 公共卫生相关性：在这里，我们提出了一种针对细菌生物膜基质特定区域的分泌蛋白的开发方法。这种技术在生物技术、生物工程和生物医学应用中可能是有用的，包括在整个细胞中有效地包装分泌抗原、灭活细菌疫苗和将功能性酶输送到 表面。

DESCRIPTION (provided by applicant): Our laboratory recently completed the first comprehensive proteomic analysis of a Gram-negative biofilm matrix. This work demonstrated that a subset of secreted proteins is retained in the biofilm matrix. Three proteins that play a structural role in the biofilm were studied in detail. Interestingly, these proteins were found to have unique distributions in the biofilm matrix that correspond to their functions in biofilm formation. The functions and distributions of several more putative biofilm-associated proteins remain to be characterized. We hypothesize that we can harness these biofilm matrix-associated proteins for use in targeting functional enzymes or antigens to specific regions of the biofilm matrix. Here we propose to develop a toolbox of biofilm matrix association modules and then to use this toolbox in proof of principle experiments in which we associate the secreted enzyme chitinase or the B subunit of cholera toxin with the biofilm matrix in specific distributions. Our goal is to establis a novel strategy for association of proteins with the biofilm matrix of Gram-negative bacteria for use in biomedical, biotechnical, and bioengineering applications of Gram-negative bacteria for use in biomedical, biotechnical, and bioengineering applications. PUBLIC HEALTH RELEVANCE: Here we propose the development of a method for targeting secreted proteins to specific regions of the bacterial biofilm matrix. Such a technology could be useful in biotechnical, bioengineering, and biomedical applications including efficient packaging of secreted antigens in whole cell, killed bacterial vaccines and delivery of functional enzymes to surfaces.