Genomics-Accelerated Natural Product Discovery

本课题组主要研究天然产物的发现、生物合成、结构和功能。 是小分子药物的最大化学物质来源。最高产的药用植物生产商 相关的NPs来源于一组被称为放线菌的细菌。回溯基因组分析已经 反复证明，具有合成能力的微生物，包括放线菌，能够合成 比目前已知的NPs要多得多。探索未被发现的大多数，即所谓的NP“暗物质”， 拥有巨大的潜力，不仅可以通过扩大我们的药物储备来改善人类健康，而且 鉴于纳米粒子作为化学探针的关键作用，也是为了推进生物学的基础研究。 该提案概述了一种以化学为中心的NP发现战略，该战略专门为解决传统的NP发现策略而设计。 传统上的缺陷。尽管传统的NP发现方法取得了成功，但最流行的生物检测- 引导性隔离存在固有的偏见，导致不可接受的高再发现率。而不是 通过其生物活性来追踪NP，我们选择通过存在以下有机官能团来识别NP 可以化学选择性地衍生，而不需要提纯。对25万个核动力源的回顾 《NP词典》显示，约50%的NP含有这样的官能团。我们将使用pmoL检测这些组 在与合适的探针反应后，在粗细菌提取物的背景下使用质谱仪进行灵敏度测定。 重要的是，我们所针对的官能团的存在可以基于生物信息预测 已知的生物合成途径。我们将这种NP发现过程称为基于反应性的筛查(RBS) 最近报道了通过亲核1，4-加成以及 非核糖体多肽(NRPs)通过肟基连接。一个定制的生物信息学软件程序也已经 由我们小组开发，用于帮助预测各种目标有机官能团的存在。 利用我们内部收集的约10,000株放线菌，其中许多来自稀有的、未被研究的分类群，我们已经去- 监督这个项目由三个独立但相互关联的目标组成。对于Aim I来说，重点是小说 硫肽，这是一种广泛的翻译后修饰多肽，以其阻断能力而闻名 细菌蛋白质翻译。AIM II重点介绍了新型多酮化合物的发现和表征 III是针对核反应堆的。每个目标都将使用多层次策略来阐明NP的结构和生物活性。 后两个目的还引入了除了已经提到的反应之外的新的探针化学。 这个我们有大量支持性数据的项目的中心假设是NP的发现 可以通过基因组学优先排序和苏格兰皇家银行的结合来加速。我们设想了这种方法 随着更多的基因组出现在公共数据库中，这将是高度可行的，并增加力量。

Our group focuses on the discovery, biosynthesis, structure, and function of natural products (NPs), which rep- resent the largest source of chemical matter for small molecule drugs. The most prolific producers of medicinally relevant NPs derive from a group of bacteria known as the actinomycetes. Retrospective genome analysis has repeatedly shown that synthetically talented microbes, including actinomycetes, are capable of synthesizing many more NPs than are currently known. Exploring the undiscovered majority, this so-called NP “dark matter”, holds enormous potential for not only improving human health by expanding our pharmaceutical repertoire but also for advancing the fundamental study of biology given the critical role of NPs as chemical probes. This proposal outlines a chemistry-centric NP discovery strategy designed specifically to address tradi- tional shortcomings. Despite the success of traditional NP discovery methods, the most prevalent, bioassay- guided isolation, suffers from inherent biases that leads to unacceptably high rates of rediscovery. Rather than track a NP by its bioactivity, we have elected to identify NPs by the presence of organic functional groups that can be chemoselectively derivatized without the need for purification. A review of the 250,000 NPs present in the Dictionary of NPs shows that ~50% contain such a functional group. We will detect these groups with pmol sensitivity using mass spectrometry in the context of crude bacterial extracts after reaction with a suitable probe. Importantly, the presence of the functional groups we are targeting can be bioinformatically predicted based on known biosynthetic pathways. We have termed this NP discovery procedure reactivity-based screening (RBS) and have recently reported the discovery of thiopeptides and polyketides by nucleophilic 1,4-addition as well as non-ribosomal peptides (NRPs) by oxime ligation. A custom bioinformatics software program has also been developed by our group to aid in predicting the presence of various targetable organic functional groups. Employing our in-house collection of ~10,000 actinomycetes, many from rare, understudied taxa, we have de- vised this project to consist of three independent but interconnected aims. For Aim I, the focus is on novel thiopeptides, which are extensively posttranslationally modified peptides best known for their ability to block bacterial protein translation. Aim II focuses on the discovery and characterization of novel polyketides while Aim III is directed towards NRPs. Each aim will elucidate NP structure and bioactivity using a multi-tiered strategy. The latter two aims also introduce new probe chemistry beyond the reactions already mentioned. The central hypothesis of this project, for which we have a large amount of supportive data, is that NP discovery can be accelerated through the marriage of genomics prioritization and RBS. We envision this approach to be highly enabling and increase in power as more genomes appear in public databases.