Bioinformatic Guided Discovery and Characterization of New RiPP Natural Products

项目总结 从青霉素到青蒿素，天然产物在这一发现和 开发新药。为了补充传统的基于活动的分离方法，基因组挖掘 通过专门针对有前景的目标，技术可以实现高效和高吞吐量的发现管道 生物体和基因簇的详细研究。然而，对生物合成途径的详细理解是 通常需要，这极大地限制了搜索过程。因此，替代的生物信息学方法 结合详细的生化特征和生物活性研究将有助于扩大我们的集体 了解自然界中具有生物活性的次生代谢物及其对人类健康的价值。 Chekan实验室的总体重点是利用非酶依赖的基因组挖掘作为指南，以实现 发现和详细研究新型生物活性天然产物和生物合成途径。这 提案的重点是核糖体合成和翻译后修饰多肽(RIPP)家族 天然产品。RIPP是一种无处不在的天然产品家族，存在于生活的各个领域，并拥有 广泛的生物活性，其中抗生素活性被特别好地表现出来。这些天然产品 它们都有一致的生物合成逻辑，但可以由各种各样的生物合成酶产生。在……里面 为了发现全新的RIPP类，我们建议利用保守的生物合成和结构 特征，而不是特定的酶转化。这将允许快速在电子计算机上识别有希望的 生物合成的基因簇，以前从未产生过孤立的分子。充分研究和描述这些 将采用生物化学和结构方法相结合的新途径。这些结果将给出 对新的生物合成酶的重要见解，揭示它们的机制，并评估它们对 工程学。最后，这个第一个项目将揭示新发现的天然产物的生物活性及其 作用机制。为了补充这一发现元素，第二个项目将寻求将多肽自然 没有生物合成信息的产品系列，以及它们的生物合成酶。这项建议 还将使培养研究生和博士后，负责下一代科学家 促进人类健康。总体而言，这一提议的结果不仅将导致发现新的天然 产品和酶，但将为未来基于基因组挖掘的发现研究和途径工程提供信息 努力寻找有希望的药物线索。

PROJECT SUMMARY Ranging from penicillin to artemisinin, natural products have been essential in the discovery and development of new drugs. To complement the traditional activity-guided isolation approaches, genome mining techniques can enable an efficient and high throughput discovery pipeline by specifically targeting promising organisms and gene clusters for detailed study. However, detailed understanding of a biosynthetic pathway is typically needed, significantly constraining the search process. Therefore, alternative bioinformatic approaches combined with detailed biochemical characterization and bioactivity studies will help expand our collective understanding of bioactive secondary metabolites in nature and their value for human health. The overall focus of the Chekan lab is to utilize enzyme-independent genome mining as a guide to enable the discovery and detailed study of new types of bioactive natural products and biosynthetic pathways. This proposal focuses on the ribosomally synthesized and post-translationally modified peptide (RiPP) family of natural products. RiPPs are a ubiquitous family of natural products found across all domains of life and possess a wide range of bioactivities, with antibiotic activity being particularly well represented. These natural products all share a consistent biosynthetic logic, but can be generated from a wide variety of biosynthetic enzymes. In order to discover completely new classes of RiPPs, we propose to exploit conserved biosynthetic and structural features instead of specific enzymatic transformations. This will allow for rapid in silico identification of promising biosynthetic gene clusters that generate never before isolated molecules. To fully study and characterize these new pathways, a combination of biochemical and structural approaches will be employed. These results will give important insights into the new biosynthetic enzymes, reveal their mechanisms, and assess their applicability for engineering. Finally, this first project will reveal the bioactivity of the newly discovered natural products and their mechanism of action. To complement this discovery element, a second project will seek to link peptidic natural product families, for which no biosynthetic information is available, to their biosynthetic enzymes. This proposal will also enable the training of both graduate students and postdocs, the next generation of scientists responsible for advancing human health. Overall, the results of this proposal will not only lead to the discovery of new natural products and enzymes, but will inform future genome mining based discovery studies and pathway engineering efforts to identify promising drug leads.