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）：生物体使用化学品作为与周围环境相互作用的一种手段，科学家们利用这一事实发现了当今用于治疗疾病的绝大多数药物。盘基网柄藻是目前已知基因组中聚酮合酶（PKS）基因最丰富的植物。此外，该生物体中的某些类型的PKS与其他酶活性蛋白质结构域融合。这种独特的杂交排列非常有效地合成了参与黏菌细胞分化的聚酮分子。这对传染病的治疗具有潜在的意义，因为临床使用的大多数常见抗生素是来自土壤传播微生物的聚酮化合物-然而，阿米巴原虫尚未在这方面进行研究。我们将开始筛选大量的土源性变形虫，以用于抗生素的生产。已故的肯尼思·雷珀（Kenneth Raper）博士是社会变形虫生态学和系统发育学方面的权威，他积累了这一资源。强奸犯档案是一个社会土传阿米巴的多样化集合，代表五个不同的属，十七个不同的物种，和一千多个独特的分离物。在这个项目中，共培养的dictyostelids和他们的细菌猎物将在固体或液体培养基中生长，这些共培养的上清液将被筛选的抗菌活性使用一组已知的病原体。这些板也将在液体和固体培养基中生长。将进行生物测定以鉴定含有抗菌活性的上清液。通过使用截留分子量（5 x 103）的膜滤器，我们将评估抗生素功能是蛋白质依赖性还是小分子依赖性。将根据抗生素的推断性质设计纯化程序。我们的R21项目将通过指导已证实的抗生素发现策略来重振药物NP研究，以靶向微生物次级代谢产物的新的和有前途的来源，真核土栖阿米巴。在短期内，这项研究将提高我们的科学知识的dictyostelids的功能基因组学。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，这些细菌以产生有价值的代谢产物而闻名，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，它们具有与昨天多产的生产者相似的生态和生化特征：土壤居住的真核社会阿米巴。