Mapping the Secondary Metabolomes of Marine Cyanobacteria

绘制海洋蓝细菌的次级代谢组图

• 批准号: 9167954
• 负责人: PIETER C DORRESTEIN
• 金额: $ 3.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-05 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9167954
• 关键词: Agrochemicals; Algorithms; Analytical Chemistry; Aquaculture; Architecture; Area; Bacteria; Biochemical; Biochemistry; Biological; Biological Assay; Biomedical Research; Breathing; Chemicals; Chromatography; Clinical Trials; Collection; Complex; Cryptophycin; Curacin A; Cyanobacterium; Cyclic Peptides; Data; Detection; Development; Dolastatin 10; Effectiveness; Environment; Family; Future; Gene Cluster; Gene Expression; Genes; Genetic Markers; Genome; Genomics; Genotype; Goals; Health; High Pressure Liquid Chromatography; Image; Inflammation; Informatics; Invertebrates; Investigation; Knowledge; Lead; Libraries; Life; Maps; Marines; Mass Spectrum Analysis; Metabolism; Methodology; Methods; Molecular; Molecular Structure; Natural Product Drug; Natural Products; Nitrogen; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Physiology; Play; Porifera; Process; Production; Prokaryotic Cells; Property; Research; Role; Sampling; Science; Series; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure; Taxon; Techniques; Technology; Therapeutic; Toxic effect; Urochordata; analog; analytical method; base; bioactive natural products; biological systems; cancer cell; cost; drug discovery; genome sequencing; improved; innovation; innovative technologies; insight; interest; mathematical algorithm; member; metabolome; microorganism; neuroregulation; novel; novel therapeutics; research study; scale up; success

DESCRIPTION (provided by applicant): Key to the process of pharmaceutical lead compound discovery from natural sources is the effective access and characterization of highly diverse molecular structures. In this regard, exploration of the marine environment for bioactive natural products is revealing new vistas in natural products chemical diversity. In this application we propose the development of innovative technologies and knowledge, principally based on LC-MS/MS data and 'molecular mapping', which will improve the effectiveness of natural products drug discovery efforts. This will enable a much improved capacity to discover new molecular diversity or analogs in desired structure classes. We will develop an understanding of the degree of expression of natural product pathways in cultured strains, and will develop novel methods by which to upregulate low or non-expressing biosynthetic gene clusters. As a result of these studies, new marine cyanobacterial natural products will be discovered and their biomedical properties will be characterized. To accomplish these goals we have the following four specific aims: 1) To use LC-MS/MS profiling of cyanobacterial extracts and pure compounds, followed by molecular mapping, to create a representation of the chemical universe of our samples. 2) To use QPCR and genome sequencing technologies to evaluate the degree of expression of natural product pathways in our cultured marine cyanobacteria, and to connect Natural Product Super-producing strains of cyanobacteria with their genotypes. This latter information can be used to find genetic markers that can be rapidly deployed to locate this phenotype in new cyanobacterial cultures and collections. 3) To use a suite of imaginative methods to transcriptionally activate cryptic natural product biosynthetic gene clusters in strains determined in Aim 2 to possess un-expressed natural products capacity, and to analyze the resulting elicited secondary metabolomes by mass spectrometry and molecular mapping. 4) To isolate members of new families of compounds detected in Aims 1, as well as newly expressed natural products from Aim 3, and rigorously establish molecular structures using advanced analytical methods. Through the course of these four specific aims, this collaborative group will explore a number of innovative methods and approaches in the natural products sciences, including MS/MS molecular mapping, genomic analysis of natural products expression, elicitation of new natural products expression, connection of natural product-rich phenotypes to their corresponding genotypes, imaging mass spectrometry of complex consortiums of species wherein natural product pathways are activated, and novel automated MS approaches to natural products characterization. All of these methods are focused on improving the detection and characterization of the molecular diversity present in microorganisms, in this case, marine cyanobacteria. This molecular diversity continues to be an important source of inspirational molecules for biomedical research and drug discovery.

描述（由申请人提供）：从天然来源发现药物铅复合过程的关键是高度多样化分子结构的有效访问和表征。在这方面，探索生物活性天然产品的海洋环境正在揭示天然产品化学多样性中的新景色。在此应用中，我们建议开发创新技术和知识，主要基于LC-MS/MS数据和“分子映射”，这将提高天然产品药物发现工作的有效性。这将使在所需的结构类别中发现新的分子多样性或类似物的能力大大提高。我们将对培养菌株中天然产物途径的表达程度发展，并将开发出上调低或不表达生物合成基因簇的新方法。这些研究的结果是，将发现新的海洋蓝细菌天然产物，并将其生物医学特性进行表征。为了实现这些目标，我们具有以下四个特定目的：1）使用蓝细菌提取物和纯化合物的LC-MS/MS分析，然后进行分子映射，以创建样品化学宇宙的表示。 2）使用QPCR和基因组测序技术评估我们培养的海洋蓝细菌中天然产物途径的表达程度，并将自然产物超产物的蓝细菌菌株与其基因型联系起来。后一种信息可用于查找可以迅速部署的遗传标记，以在新的蓝细菌培养物和收集中找到这种表型。 3）使用一套富有想象力的方法在菌株中转录激活隐性天然产物生物合成基因簇 在AIM 2中确定具有未表达的天然产物容量，并通过质谱和分子映射分析所得的次级代谢组。 4）隔离AIM 1中检测到的新化合物的成员以及AIM 3的新表达天然产品，并使用先进的分析方法严格建立分子结构。通过这四个特定目的的过程，这个协作小组将探索自然产品科学中的许多创新方法和方法，包括MS/MS分子映射，对天然产物表达的基因组分析，新的天然产物表达的基因组分析，自然产物表达的连接，自然产物表型的连接与它们的相应基因型和相应的质量质量生产的综合产品的相关方法，该物种是自然的质谱，该物种的复杂性是一种物种的复杂性，而这些方法是在其中的种类，其中包括物种的复杂性，这些物种是一种物种的复杂性，而这些方法是在其中，其中的物种是构成物种的综合产品。天然产品表征。所有这些方法都集中在改善微生物中存在的分子多样性的检测和表征，在这种情况下是海洋蓝细菌。这种分子多样性仍然是生物医学研究和药物发现的励志分子的重要来源。