Mapping the Secondary Metabolomes of Marine Cyanobacteria

绘制海洋蓝细菌的次级代谢组图

• 批准号: 8562582
• 负责人: PIETER C DORRESTEIN
• 金额: $ 45.24万
• 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 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8562582
• 关键词: Agrochemicals; Algorithms; Analytical Chemistry; Aquaculture; Architecture; Area; Bacteria; Biochemical; Biochemistry; Biological; Biological Assay; Biological Factors; 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; 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; 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; biological systems; cancer cell; cost; drug discovery; genome sequencing; improved; innovation; innovative technologies; insight; interest; mathematical algorithm; member; microorganism; neuroregulation; novel; public health relevance; 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先导化合物的过程的关键是有效获取和表征高度多样化的分子结构。在这方面，对海洋环境中生物活性天然产物的探索正在揭示天然产物化学多样性的新前景。在此应用中，我们建议开发主要基于 LC-MS/MS 数据和“分子图谱”的创新技术和知识，这将提高天然产物药物发现工作的有效性。这将大大提高发现所需结构类别中的新分子多样性或类似物的能力。我们将了解培养菌株中天然产物途径的表达程度，并将开发上调低表达或非表达生物合成基因簇的新方法。这些研究的结果将是发现新的海洋蓝藻天然产物，并表征其生物医学特性。为了实现这些目标，我们有以下四个具体目标：1) 使用 LC-MS/MS 对蓝藻提取物和纯化合物进行分析，然后进行分子图谱分析，以创建我们样品的化学宇宙的表示。 2）利用QPCR和基因组测序技术评估我们培养的海洋蓝藻中天然产物途径的表达程度，并将蓝藻天然产物超产菌株与其基因型联系起来。后一信息可用于寻找遗传标记，这些标记可快速部署以在新的蓝藻培养物和收藏品中定位该表型。 3）使用一套富有想象力的方法来转录激活菌株中神秘的天然产物生物合成基因簇 目标 2 中确定其具有未表达的天然产物能力，并通过质谱和分子作图分析所产生的次级代谢组。 4) 分离目标1中检测到的新化合物家族的成员，以及目标3中新表达的天然产物，并使用先进的分析方法严格建立分子结构。通过这四个具体目标的课程，该合作小组将探索天然产物科学中的许多创新方法和途径，包括MS/MS分子图谱、天然产物表达的基因组分析、新天然产物表达的诱导、天然产物丰富的表型与其相应的基因型，其中天然产物途径被激活的复杂物种联合体的成像质谱分析，以及天然产物表征的新型自动化 MS 方法。所有这些方法都集中于改进微生物（在本例中为海洋蓝藻）中存在的分子多样性的检测和表征。这种分子多样性仍然是生物医学研究和药物发现的灵感分子的重要来源。