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，用于开发第二种 二茂铁基抗真菌萜类化合物的合成。我们的团队由Mead(Varigen)，Keller(University of 威斯康星)和凯莱赫(西北大学)利用 将基因组学与真菌萜类合成的代谢组学理解相结合的技术融合。我们 首先，应用基因组挖掘驱动的方法来寻找新的天然恩福马芬净类似物。由此产生的 然后，将数据与代谢组分析和抗真菌生物活性分析数据相结合，以建立假定的 不同菌株编码的Enfumafungin类代谢物，并最终绘制结构-活性关系图 在这类有效的抗真菌药物中。同时，我们的团队正在构建一个模块化平台，能够 生成数百种不同的功能数量的类似物。在这里，我们将异源地表达 工程化尼杜拉曲霉宿主中真菌菌株的Enfumafungin-like通路。序列- 在该计划中发现的铅分子的特异性克隆和异源表达将建立方法 在多个物种之间进行有针对性的环化酶基因交换，以产生新的抗真菌铁烯类化合物 脚手架。需要额外资源的长期目标是推动第二代三萜类化合物的发展 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步 朝着建立功能代谢基因组学作为从基因组到功能的可行工具迈出一步 有治疗作用。作为试验床，我们转向一种具有最近临床实用历史的天然产品系列 开发新的类似物和获取它们的技术。

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.