Discovery and characterization of new natural products from uncultured bacteria

来自未培养细菌的新天然产物的发现和表征

• 批准号: 7683813
• 负责人: SEAN F BRADY
• 金额: $ 32万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683813
• 关键词: Address; Anabolism; Archives; Aromatic Amino Acids; Bacteria; Biological; Biological Assay; Biological Factors; Biological Process; Collection; Conserved Sequence; Cosmids; Costa Rica; Coupled; DNA; DNA Library; Data; Environment; Enzymes; FDA approved; Gene Cluster; Genes; Genetic Variation; Glycerol; Lactams; Libraries; Location; Microbe; Molecular; Nature; Pathway interactions; Peptides; Phylogenetic Analysis; Production; Protocols documentation; Reaction; Recovery; Research Personnel; Running; Sampling; Screening procedure; Site; Soil; Source; Tanzania; Testing; Therapeutic; Therapeutic Agents; Thiazoles; United States; base; cellular targeting; chemical genetics; complex biological systems; human disease; method development; microbial host; novel; programs; soil sampling

Project Summary: The study of biologically active natural products has been a very effective gateway to the identification of compounds with unique cellular targets that have been useful both as therapeutics and,more recently in the emerging field of chemical genetics, as modulators of complex biological systems. One of the key revelations to come from molecular phylogenetic analyses of environmental samples is that only a small fraction of bacteria present in the environment is easily cultured. Traditional approaches used for the discovery of new natural products are therefore unlikely to provide access to the vast majority of natural products that exist in nature. Soil microbes that have not yet been cultured and as a result have not been examined for the production of useful natural products outnumber their cultured counterparts by at least two to three orders of magnitude. Using PCR with degenerate primers based on conserved regions in genes known to be involved in the biosynthesis of natural products and high throughput sequencing we are proposing to screen cosmid libraries of DNA extracted directly from soil for biosynthetic pathways that encode the production of new natural products. Heterologous expression will then be used to access the molecules encoded by these new gene clusters and the resulting molecules screened for biological activity in a variety of human disease related screens. Relevance: Natural products, naturally derived small organic compounds, have traditionally been one of the major sources of FDA approved therapeutics. Uncultured bacteria are one of the largest remaining pools of genetic diversity that have not yet been examined for the production of secondary metabolites. The development of methods to functionally access the previously inaccessible natural products produced by uncultured bacteria should significantly increase the number and diversity of natural products that are available to test as probes of biological processes and therapeutic agents.

项目概要： 生物活性天然产物的研究已成为鉴定天然产物的非常有效的途径。 具有独特细胞靶点的化合物既可用作治疗药物，也可用于最近的研究 化学遗传学的新兴领域，作为复杂生物系统的调节剂。关键之一 来自环境样本的分子系统发育分析的启示是，只有一小部分 环境中存在的部分细菌很容易培养。传统方法用于 因此，新天然产品的发现不太可能提供绝大多数天然产品的获取途径。 自然界中存在的产品。尚未培养且因此尚未被处理的土壤微生物 经过检查，有用的天然产品的生产数量至少比人工培养的产品多两倍 到三个数量级。使用基于基因保守区域的简并引物进行 PCR 已知参与天然产物的生物合成和高通量测序，我们是 提议筛选直接从土壤中提取的 DNA 粘粒文库，以寻找生物合成途径 对新天然产品的生产进行编码。然后，异源表达式将用于访问 这些新基因簇编码的分子以及筛选所得分子的生物活性 各种人类疾病相关屏幕。 关联： 天然产物，天然衍生的小有机化合物，传统上是主要的产品之一 FDA 批准的治疗药物的来源。未培养的细菌是最大的剩余细菌库之一 尚未检查次生代谢物产生的遗传多样性。这 开发方法以功能性地获取以前无法获取的天然产物 未培养的细菌应该显着增加天然产物的数量和多样性 可作为生物过程和治疗剂的探针进行测试。