Collaborative Proposal: Biosynthesis of Alkamides - Experimental Modeling of a Modular Secondary Metabolic Pathway

智力优点：烷酰胺（与脂肪酸连接的酰胺）是新型植物次生化合物，其特征很少，但在紫锥菊中积累到显着水平，使得这种植物成为研究这些化合物及其生物合成的选择。我们对植物中烷酰胺功能的了解还处于起步阶段，但最近的数据表明，除了杀虫特性外，这些化合物可能还具有新的信号传导功能。这项工作将为进一步研究这些化合物在植物生物学中的重要性奠定基础，并开启对这种已药用数百年的植物属的研究。由于烷酰胺生物合成途径涉及氨基酸代谢和新颖的化学键，因此所涉及的酶在未来也有多种生物技术应用。拟议的研究将战略性地应用高通量全局分析技术来阐明这种天然产物的生物合成途径。 烷酰胺似乎是通过模块化代谢途径进行生物组装的，该途径可能是氨基酸和脂肪酸代谢的适应。 结合代谢物分析、代谢流研究和转录组学的实验将用于鉴定组装多种烷酰胺拼贴画的基因和酶。 定义烷酰胺途径提供了发现新的代谢过程的潜力，这些过程产生新的化学功能组合，具有广泛的应用（例如，润滑和洗涤剂行业）。 此外，该研究项目概述了一种通用方法，该方法应广泛适用于发现初级和特化植物代谢如何并置和进化以产生植物分类群之间的理化表型差异。 所使用的多层生物勘探提供了浏览生物体代谢库的机会，所涉及生化过程的全系统知识应转化为与化学工业相关的新型生物衍生化合物的创造，以及害虫抗性策略。 更广泛的影响：该项目将促进 IUPUI、ISU 研究人员和美国农业部中北部地区植物引种站的科学家之间的多学科伙伴关系。为开展这项研究而建立的合作伙伴关系将通过本科生、研究生和博士后的男女同校，在有机化学、机械生物化学和功能基因组学日益罕见的交叉领域为年轻科学家提供指导。 IUPUI 是一所大型城市大学，其学生将从麦克奈尔和多元化学者 STEM 研究项目的实习生库中选拔，其中有大量少数族裔和第一代学生。这些互动将使该项目能够与 IUPUI 校园计划协同运作，指导本科生接受研究生教育。作为 IUPUI 特有的推广活动，高中教师将体验实践研究。他们将与研究生一起进行实验，从而获得适合在其家庭学校使用的生物化学课程。这项工作与 NSF GK-12 持续参与 IUPUI 的研究项目相吻合。在 ISU，本科生将把拟议的研究转化为生物技术生物化学新课程的一个模块。

Intellectual Merit: Alkamides (amides linked to fatty acids) are novel plant secondary compounds that are poorly characterized, but that accumulate to significant levels in Echinacea, making this plant of choice to study these compounds and their biosynthesis. Our understanding of alkamide function in plants is in its infancy, but recent data suggest these compounds may have novel signaling functions in addition to insecticidal properties. This work will lay the foundation for further studies of the importance of these compounds in plant biology as well as open up research on this plant genus which has been used medicinally for hundreds of years. Because the pathways in alkamide biosynthesis involve amino acid metabolism and novel chemical linkages, there are also multiple future biotechnological applications for the enzymes involved. The proposed research will strategically apply high-throughput global profiling technologies to elucidate this natural product biosynthetic pathway. Alkamides appear to be biologically assembled via a modular metabolic pathway that may be an adaptation of amino acid and fatty acid metabolism. Experiments that combine metabolite profiling, metabolic flux studies and transcriptomics will be used to identify genes and enzymes that assemble a diverse collage of alkamides. Defining the alkamide pathway offers the potential of discovering new metabolic processes that generate novel combinations of chemical functionalities, which have wide-ranging applications (e.g., lubrication and detergent industries). In addition, this research project outlines a general methodology that should be broadly applicable to discovering how primary and specialized plant metabolism are juxtaposed and evolve to generate the physiochemical phenotypic differences among plant taxonomic groups. The multilayered bio-prospecting to be used offers the opportunity to browse the metabolic repertoire of an organism, and the system-wide knowledge of the involved biochemical processes should translate to the creation of novel bio-derived compounds relevant to the chemical industries, as well as strategies for pest resistance.Broader Impacts: This project will facilitate a multidisciplinary partnership between IUPUI, ISU researchers and scientists at the USDA North Central Regional Plant Introduction Station. The collaboration established to conduct this research will mentor young scientists at the increasingly uncommon intersection of organic chemistry, mechanistic biochemistry and functional genomics through the coeducation of undergraduates, graduate students and post-doctoral fellows. Students at IUPUI, a large, urban university, will be drawn from a trainee pool rich in underrepresented minority and first generation students in the McNair and Diversity Scholars STEM research programs. These interactions will allow the project to operate synergistically with IUPUI campus initiatives to mentor undergraduate students toward graduate education. As an outreach activity specific to IUPUI, high school teachers will experience hands-on research. In conjunction with the graduate students, they will engage in experiments leading to biochemical lessons suitable for use at their home schools. This effort dovetails with ongoing NSF GK-12 participation in IUPUI's research programs. At ISU, undergraduates will translate the proposed research into a module in a new course in biotechnological biochemistry.