Mining a collection of soil amoebae for antibacteria agents

挖掘土壤阿米巴原虫群以获取抗菌剂

• 批准号: 8264540
• 负责人: MARCIN S FILUTOWICZ
• 金额: $ 18.17万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8264540
• 关键词: Amoeba genus; Anti-Bacterial Agents; Anti-Infective Agents; Antibiotic Resistance; Antibiotics; Archives; Back; Bacteria; Bacteriology; Biochemical; Biological Assay; Biological Factors; Biological Response Modifier Therapy; Biology; Cells; Characteristics; Chemical Structure; Chemicals; Clinical; Coculture Techniques; Collection; Development; Dictyostelium; Dictyostelium discoideum; Disease; Ecology; Enzymes; Exhibits; Foundations; Freezing; Funding; Genome; Genomics; Growth; High Pressure Liquid Chromatography; Hybrids; Investigation; Ion Exchange Resins; Island; Knowledge; Lead; Learning; Liquid substance; Literature; Membrane; Methods; Microbe; Mining; Molecular Weight; Natural Products Chemistry; Nature; Nuclear Magnetic Resonance; Organic solvent product; Organism; Phagocytes; Pharmacologic Substance; Phylogeny; Physarum polycephalum; Procedures; Process; Production; Proteins; Relative (related person); Research; Resources; Scientist; Screening procedure; Secondary to; Soil; Solid; Solubility; Solvents; Source; Staging; System; Tertiary Protein Structure; Therapeutic; Thin Layer Chromatography; Universities; Wisconsin; Work; antimicrobial; authority; base; design; drug discovery; functional genomics; improved; infectious disease treatment; interest; microbial; microorganism; novel; pathogen; pathogenic bacteria; polyketide synthase; public health relevance; repository; research study; small molecule; social

DESCRIPTION (provided by applicant): Organisms use chemicals as a means of interacting with their surroundings and scientists have exploited this fact to discover the vast majority of pharmaceuticals used to treat disease today. Dictyostelium discoideum exhibits the largest repository of polyketide synthase (PKS) enzymes of all known genomes. Furthermore, some types of PKSs in this organism are fused to other enzymatically active protein domains. The unique hybrid arrangement works with great efficiency to synthesize polyketide molecules that are involved in the differentiation of slime mold cells. This has potential implications for the treatment of infectious disease because most of the common antibiotics in clinical use are polyketides derived from soil-borne microbes - however, amoebae have not been examined in this regard. We will begin screening a large archive of soil- borne amoebae for the production of antibiotics. The late Dr. Kenneth Raper, an authority on the ecology and phylogeny of social amoebae, amassed this resource. The Raper Archive is a diverse collection of social soil- borne amoebae representing five different genera, seventeen different species, and over a thousand unique isolates. In this project, co-cultures of dictyostelids and their bacterial prey will be grown in solid or liquid media and the supernatants of these co-cultures will be screened for antimicrobial activity using a panel of known pathogens. The panels will also be grown both in liquid and on solid media. Bioassays will be conducted to identify supernatants that contain antibacterial activity. By using membrane filters with a molecular weight cutoff (5 x 103), we will assess whether the antibiotic function is proteinaceous or small molecule-dependent. Purification procedures will be designed according to the inferred nature of the antibiotic. Our R21 project will reinvigorate pharmaceutical NP research by directing proven antibiotic discovery strategies to target a new and promising source of microbial secondary metabolites, eukaryotic soil-dwelling amoebae. In the short term, this research will improve our scientific knowledge of the functional genomics of dictyostelids. And ultimately, our work will identify lead compounds for much-needed antibiotics that may have novel chemical structures and mechanisms of action. PUBLIC HEALTH RELEVANCE: The growing problem of antibiotic resistance is generating a need for vigorous drug discovery research. Most clinical antibiotics come from screens of soil-dwelling bacteria, which are renowned for their production of valuable metabolites, but are increasingly yielding previously discovered antibiotics. In this antibiotic discovery project we will screen an untapped group of microorganisms with similar ecological and biochemical characteristics to the prolific producers of yesterday: the soil-dwelling, eukaryotic social amoebae.

描述（由申请人提供）：生物体使用化学物质作为与周围环境相互作用的手段，科学家利用这一事实发现了当今用于治疗疾病的绝大多数药物。盘基网柄菌拥有所有已知基因组中最大的聚酮合酶 (PKS) 库。此外，该生物体中的某些类型的 PKS 与其他酶活性蛋白结构域融合。独特的混合排列可以非常有效地合成参与粘菌细胞分化的聚酮化合物分子。这对传染病的治疗具有潜在的影响，因为临床使用的大多数常见抗生素都是源自土传微生物的聚酮化合物 - 然而，阿米巴原虫尚未在这方面进行检查。我们将开始筛选大量用于生产抗生素的土传变形虫。已故社会变形虫生态学和系统发育学权威肯尼思·雷珀博士积累了这一资源。 Raper Archive 是社会性土传变形虫的多样化收藏，代表 5 个不同属、17 个不同物种和 1000 多个独特的分离株。在该项目中，网柄菌及其细菌猎物的共培养物将在固体或液体培养基中生长，并且将使用一组已知病原体筛选这些共培养物的上清液的抗菌活性。这些面板还将在液体和固体培养基上生长。将进行生物测定以确定含有抗菌活性的上清液。通过使用截留分子量 (5 x 103) 的膜过滤器，我们将评估抗生素功能是否依赖于蛋白质或小分子。纯化程序将根据抗生素的推断性质来设计。我们的 R21 项目将通过指导经过验证的抗生素发现策略，以微生物次生代谢物的新的、有前途的来源——真核土壤阿米巴原虫为目标，重振药物 NP 研究。从短期来看，这项研究将提高我们对网柄科动物功能基因组学的科学认识。最终，我们的工作将确定急需的抗生素的先导化合物，这些化合物可能具有新颖的化学结构和作用机制。 公共卫生相关性：日益严重的抗生素耐药性问题正在产生对积极药物发现研究的需求。大多数临床抗生素来自土壤细菌的筛选，这些细菌以其产生有价值的代谢物而闻名，但越来越多地产生以前发现的抗生素。在这个抗生素发现项目中，我们将筛选一组未开发的微生物，它们与昨天的多产生产者具有相似的生态和生化特征：土壤栖息的真核社会变形虫。