Mechanistic Study of Cellulosome Through Reprogramming Its Assembly

小行星1264708 本研究旨在研究纤维素体的自组装过程，纤维素体是一组由微生物分泌的天然蛋白质复合物，用于高效降解植物细胞壁为可发酵糖。 多纤维素酶体是由一个具有结构功能的核心支架蛋白和几个具有催化功能的水解蛋白组成。 多纤维素体的组装通过支架蛋白的粘附蛋白结构域和催化模块的匹配锚定蛋白结构域之间的相互作用发生。 由于在单一微生物中粘着蛋白-锚定蛋白识别的非歧视性性质，组装的纤维素体具有不同的结构和分子组成。 拟议的研究旨在实现区域和化学计量控制的设计师cellulosomes通过工程正交cohesin-dockerin蛋白对最小的crosstalking识别。 具有改变的特异性的工程化的粘附素结构域将在预先排列的位置组装以形成重组支架蛋白，其能够通过匹配的锚定蛋白结构域模板化连接催化模块以形成设计的纤维素体。 我们将研究它们的构效关系，并期望通过这些研究，合成出在植物纤维素材料降解过程中性能优于天然纤维素体的纤维素体。 它将有助于产生生物质衍生原料，用于生物燃料和生物能源生产，这有可能解决许多长期的经济和环境问题。 此外，对多纤维素体自组装的速度和精度的更深入了解可能会导致自组装蛋白质复合物合成的新策略，对生物催化，代谢工程，合成生物学和纳米科学领域产生广泛影响。拟议的研究为各个发展水平的学生提供了出色的培训机会。 学生将深入接触内布拉斯加大学化学和生物学的大量研究问题。 一些具体的推广活动包括为四年制大学的本科生制定暑期研究计划和为高中生制定化学实验室工具包。 这些活动将吸引大学生和高中生，特别是妇女和代表性不足的学生，在STEM领域从事研究和学术事业。该项目由化学、生物工程、环境和运输系统司的生物技术、生物化学和生物质工程项目，分子和细胞生物学司的系统和合成生物学项目，刺激竞争研究实验计划办公室（EPSCoR）。

1264708 Guo, Jiantao This research seeks to study the self-assembly process of cellulosomes, a group of natural protein complexes secreted by microorganisms for the highly efficient degradation of plant cell walls into fermentable sugars. A cellulosome consists of a core scaffoldin protein of structural function and several hydrolytic proteins of catalytic function. Assembly of the cellulosome occurs via interactions between cohesin domains of the scaffoldin protein and matching dockerin domains of catalytic modules. Due to the non-discriminatory nature of cohesin-dockerin recognition in a single microorganism, the assembled cellulosomes have diverse structural and molecular composition. The proposed research seeks to achieve regio- and stoichiometric controlled assembly of designer cellulosomes through engineering orthogonal cohesin-dockerin protein pairs with minimal crosstalking recognition. The engineered cohesin domains with altered specificity will be assembled at pre-arranged positions to form recombinant scaffoldin protein, which enables templated attachment of catalytic modules through matching dockerin domains to form designer cellulosomes. Their structure-activity relationships will be investigated.The proposed research is expected to lead to synthetic cellulosomes that can outperform their natural counterparts in the process of plant cellulosic material degradation. It will contribute to the generation of biomass-derived feedstocks for biofuel and bioenergy production, which has potential to resolve many long-term economic and environmental concerns. Furthermore, a deeper understanding of the speed and precision of cellulosome self-assembly could lead to new strategies for the synthesis of self-assembled protein complexes with broad impact on the fields of biocatalysis, metabolic engineering, synthetic biology, and nanosciences.The proposed research represents an outstanding training opportunity for students at all levels of development. Students will be exposed in depth to a large array of research problems in chemistry and biology at the University of Nebraska. Some specific outreach activities include development of a summer research program for undergraduate students from four-year colleges and of chemistry lab kits for high school students. These activities will attract college and high school students, including specifically women and underrepresented students, to pursue studies and academic careers in STEM areas.This project is jointly funded by the Biotechnology, Biochemical and Biomass Engineering Program of the Chemical, Bioengineering, Environmental, and Transport Systems Division, by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology, and by the Office of the Experimental Program to Stimulate Competitive Research (EPSCoR).