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

智力优势：碱酰胺（与脂肪酸连接的酰胺）是一种新型的植物次生化合物，其特征很差，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。我们对植物中烷酰胺功能的理解还处于起步阶段，但最近的数据表明，这些化合物除了杀虫特性外，还可能具有新的信号功能。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，并开辟对这种已被药用数百年的植物属的研究。由于在醇酰胺生物合成的途径涉及氨基酸代谢和新的化学键，也有多种未来的生物技术应用的酶参与。拟议的研究将战略性地应用高通量全球分析技术来阐明这种天然产物的生物合成途径。 碱酰胺似乎是通过一个模块化的代谢途径，可能是一个适应氨基酸和脂肪酸代谢的生物组装。 结合联合收割机代谢物分析，代谢通量研究和转录组学的实验将被用来确定基因和酶，组装一个不同的拼贴碱酰胺。 定义烷酰胺途径提供了发现新的代谢过程的可能性，这些代谢过程产生化学官能团的新组合，这些化学官能团具有广泛的应用（例如，润滑和洗涤剂工业）。 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专业植物代谢是如何并列和进化的，以产生植物分类群之间的理化表型差异。 将采用的多层次生物勘探提供了浏览一种有机体的代谢谱的机会，而对所涉及的生物化学过程的全系统知识应转化为与化学工业有关的新型生物衍生化合物的创造，以及抗虫害的战略。该项目将促进IUPUI，ISU研究人员和美国农业部中北部区域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将通过本科生，研究生和博士后研究员的共同教育，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点指导年轻科学家。IUPUI是一所大型城市大学，其学生将来自麦克奈尔和多样性学者STEM研究项目中代表性不足的少数民族和第一代学生的培训人才库。这些互动将使该项目与IUPUI校园倡议协同运作，指导本科生进行研究生教育。作为IUPUI特有的外展活动，高中教师将体验动手研究。与研究生一起，他们将从事实验，导致适合在他们的家乡学校使用的生物化学课程。这一努力与NSF GK-12正在参与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.