Mass Spectrometric Studies of Thiotemplate Biosynthesis

硫模板生物合成的质谱研究

基本信息

批准号：
9240599
负责人：
NEIL L KELLEHER
金额：
$ 32.73万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9240599
关键词：
Actinobacteria class Amino Acids Anabolism Antibiotics Bacteria Basic Science Binding Proteins Biological Assay Chemicals Collection Communities Computer Simulation Country Data Development Exhibits Family Gene Cluster Genome Genomics Goals Grant Hand Harvest Health Hospitals Human Hybrids Investigation Joints Laboratories Left Libraries Literature Medicine Methods Microbial Genome Sequencing Mining Modeling Modus Molecular Structure Molecular Target Natural Products Nature Oxidoreductase Pharmacologic Substance Polyenes Process Productivity Proteomics Public Health Publications Research Research Personnel Resources Solid Source Stream Structure Surveys Synthesis Chemistry Work base complex biological systems drug development human disease innovation insight interdisciplinary approach metabolomics microbial microorganism monomer novel novel strategies novel therapeutics pharmacophore public health relevance screening small molecule

项目摘要

DESCRIPTION (provided by applicant): As microbial genome sequencing becomes more widespread, the capacity of microorganisms to produce natural products is coming into better view. This competing renewal surveys this capacity using the latest in computational and expression screening and targets specific sub-classes of natural products that emerge from a fundamentally new platform for discovery of natural products from the microbial world. Described here is the pursuit of specific set of natural products and their gene clusters which have emerged from a new approach that achieves large-scale pairing of biosynthetic gene clusters with their small molecules for discovery. From a library of 178 actinobacteria strains, a genome-enabled metabolomics approach was used (a.k.a., "metabologenomics") to correctly identify natural products and correlate them with their biosynthetic gene clusters. The proposed research extends this method to examine unknown metabolites, tambromycin (Aim 1) and rimosamides (Aim 2), each with their respective clusters exhibiting unusual biosynthetic mechanisms and monomers, such as the new amino acid, "tambroline" (see Aim 1). In the first two aims, the precise structures, biosynthetic details and in the case of tambromycin-the protein binding target(s) of these new natural products will be studied and defined. In Aim 3a, a set of new gene cluster-compound pairs will be mined in silico for those that terminate in reductase domains, and therefore likely harbor electrophilic warheads from reductive off-loading from thiotemplate assembly lines. Aim 3b proposes an expansion of this gene-cluster focused approach to discovering new bioactive metabolites by targeted mining of metabologenomics data for other pharmacophores known to elicit bioactivity. The specific case of a new polyene antibiotic is put forth based on sequence data indicating the presence of multiple dehydrogenase domains within the biosynthetic cluster. Mining the microbial world with "metabologenomics" represents a major shift from past activities in this R01 grant (which led to 18 publications over the past granting period). With a set of new compounds and their biosynthetic gene clusters in hand, the investigators will pursue several of these in targeted mode. Beyond the direct impact of this proposal, the work provides a major path forward for an 'omics'-driven resurgence in natural products discovery. Such a resurgence promises a more deterministic path for structure- based discovery of natural products and to provide these structures at a rate not seen before in the field (a.k.a. "high throughput discovery" as described in recent literature). In essence, the proposed research will demonstrate the value of regularizing a microbial strain collection into a bona fide library of compounds, which are known to be expressed and possess particular sub-structures (e.g., an electrophilic warhead). These value propositions serve as motivating rationales to merge a new kind of discovery pipeline containing unique molecular structures with assays for bioactivity.

描述（由申请人提供）：随着微生物基因组测序变得更加普遍，微生物生产天然产物的能力越来越受到人们的关注，这项竞争性的更新利用最新的计算和表达筛选技术来调查这种能力，并针对特定的子类。从微生物世界发现天然产物的全新平台中出现的天然产物这里描述的是对特定天然产物及其基因簇的追求，这些天然产物及其基因簇是通过实现大规模配对的新方法而出现的。从 178 个放线菌菌株的文库中，使用基因组代谢组学方法（又名“代谢基因组学”）来正确识别天然产物并将其与其生物合成基因簇相关联。将此方法扩展到检查未知代谢物坦溴霉素（目标 1）和利莫酰胺（目标 2），每种代谢物都具有它们各自的簇表现出不寻常的生物合成机制和单体，例如新的氨基酸“tambroline”（参见目标 1），在前两个目标中，精确的结构、生物合成细节以及坦溴霉素的蛋白质结合靶标（ s) 这些新的天然产物将被研究和定义，在目标 3a 中，将在计算机中挖掘一组新的基因簇-化合物对，以查找终止于还原酶结构域的化合物。因此，Aim 3b 可能含有来自硫模板装配线还原卸载的亲电子弹头，提出了一种扩展这种以基因簇为中心的方法，通过有针对性地挖掘已知能引发新生物活性的其他药效团的代谢基因组学数据来发现新的生物活性代谢物。多烯抗生素是根据表明生物合成簇内存在多个脱氢酶结构域的序列数据提出的。 “代谢基因组学”代表了 R01 资助中过去活动的重大转变（在过去的资助期间发表了 18 篇出版物），研究人员将有针对性地研究其中的一些新化合物及其生物合成基因簇。除了该提案的直接影响之外，这项工作还为“组学”驱动的天然产物发现的复兴提供了一条主要道路，这种复兴为基于结构的天然产物发现提供了一条更具确定性的道路。本质上，拟议的研究将证明将微生物菌株集合规范化为真正的库的价值。这些价值主张作为将包含独特分子结构的新型发现管道与分析相结合的激励原理。生物活性。