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

智力优势：碱酰胺(与脂肪酸相连的酰胺)是一种新的植物次生化合物，特征不佳，但在紫锥菊中积累到显著水平，使这种植物成为研究这些化合物及其生物合成的首选。我们对烷胺在植物中的功能的了解还处于起步阶段，但最近的数据表明，这些化合物除了杀虫特性外，可能还具有新的信号功能。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，并开启对这一已有数百年药用历史的植物属的研究。由于烷胺生物合成的途径涉及氨基酸代谢和新的化学键，因此所涉及的酶也有多种未来的生物技术应用。这项拟议的研究将战略性地应用高通量全球图谱技术来阐明这一天然产物生物合成途径。烷胺类化合物似乎是通过模块化的代谢途径在生物上组装的，这可能是氨基酸和脂肪酸代谢的一种适应。结合代谢物图谱、代谢流研究和转录组学的实验将被用来识别组装不同烷胺类拼贴的基因和酶。定义烷胺途径提供了发现新的代谢过程的潜力，这些过程产生了具有广泛应用的化学功能的新组合(例如，润滑和洗涤剂工业)。此外，这项研究项目概述了一种普遍适用的方法，该方法应该广泛适用于发现初级和专门的植物新陈代谢是如何并列和进化的，以产生植物分类类群之间的理化表型差异。将要使用的多层次生物勘探提供了浏览生物体代谢谱的机会，涉及的生化过程的全系统知识应该转化为与化学工业相关的新生物衍生化合物的创造，以及抗虫害的策略。广泛的影响：该项目将促进IUPUI、ISU研究人员和美国农业部北中心区域植物引种站科学家之间的多学科伙伴关系。为进行这项研究而建立的合作将通过本科生、研究生和博士后研究员的男女同校教育，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指导年轻科学家。IUPUI是一所大型的城市大学，其学生将从麦克奈尔和多元化学者STEM研究项目中代表不足的少数族裔和第一代学生中挑选出来。这些互动将使该项目与IUPUI校园倡议协同运作，指导本科生接受研究生教育。作为一项专门针对IUPUI的外展活动，高中教师将体验实践研究。他们将与研究生一起进行实验，以获得适合本国学校使用的生物化学课程。这一努力与正在进行的NSF GK-12参与IUPUI的研究计划相吻合。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.