Multi-Omics And Synbio Enabled Discovery Of Antifungal Fernene Triterpenes

项目摘要。提出了一种新的功能代谢基因组学方法， 基于蕨烯的抗真菌萜烯的产生。我们的团队由米德（瓦里根），凯勒（大学） 威斯康星州）和凯莱赫（西北大学）探索这种独特的三萜家族的多样性， 将基因组学与真菌萜烯合成的代谢组学理解结合起来的技术融合。我们 开始通过应用基因组挖掘驱动的方法来寻找恩夫马芬净的新的天然类似物。所得 然后将数据与代谢组学分析和抗真菌生物活性测定数据整合，以建立推定的 不同菌株编码的恩福霉素样代谢物，并最终绘制结构-活性关系 在这类强效抗真菌药物中占很大比例。与此同时，我们的团队正在构建一个模块化平台， 产生数百种不同的功能量的类似物。在这里，我们将异序表达 来自工程化构巢曲霉宿主中的真菌菌株的恩富霉素样途径。顺序是- 本计划中发现的先导分子的特异性克隆和异源表达将建立方法 在多个物种中进行环化酶基因的靶向交换， 脚手架长期的目标，需要额外的资源，是推进第二代三萜 抗真菌类似物进入研究和临床市场。总体而言，该计划提供了一个重要的下一个 建立功能代谢基因组学作为从基因组到功能 有治疗作用的作为一个试验平台，我们转向一个天然产品家族，该家族具有最近的临床应用历史， 开发新的类似物和技术来获取它们。

Project Summary. A new functional metabologenomics approach is proposed for the development of a second generation of fernene-based antifungal terpenes. Our team led by Mead (Varigen), Keller (University of Wisconsin) and Kelleher (Northwestern University) explore the diversity of this unique family of triterpenes using a fusion of technologies that unites genomics with metabolomic understanding of fungal terpene synthesis. We begin by applying a genome mining-driven approach to find new natural analogs of enfumafungin. The resulting data are then integrated with metabolomic analysis and antifungal bioactivity assay data to establish the putative enfumafungin-like metabolites encoded by diverse strains and ultimately chart the structure-activity relationship space within this potent class of antifungals. In parallel, our team is constructing a modular platform capable of generating hundreds of different analogs in functional amounts. Here, we will heterologously express the enfumafungin-like pathways from fungal strains in an engineered Aspergillus nidulans host. The sequence- specific cloning and heterologous expression of lead molecules discovered in this program will establish methods to conduct targeted swapping of cyclase genes across multiple species to generate new antifungal fernene-like scaffolds. The long-term goal, requiring additional resources, is to advance a second generation of triterpene antifungal analogs into the research and clinical market place. Overall, this program provides an important next step towards establishing functional metabologenomics as a viable tool to go from genome to functional therapeutic. As a testbed, we turn to a natural product family with a recent history of clinical utility for the development of new analogs and technologies to access them.