Natural Products from Under-explored Microorganisms

来自未充分探索的微生物的天然产物

• 批准号: 7758428
• 负责人: Ben Shen
• 金额: $ 37.12万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7758428
• 关键词: Actinobacteria class; Actinomyces; Bacteria; Biochemical; Biodiversity; Biological Assay; Biological Factors; Biology; Chemicals; China; Chinese People; Detection; Fermentation; High Pressure Liquid Chromatography; Human; Institutes; Libraries; Medicinal Plants; Microbiology; Molecular Bank; Pharmaceutical Preparations; Preparation; Process; Production; Screening procedure; Structure; Symbiosis; Universities; base; member; microbial; microorganism; novel; public health relevance; scale up; tool

DESCRIPTION (provided by applicant): We propose to isolate novel natural products from under-explored microorganisms and provide them to the Molecular Libraries Screening Center Network (MLSCN). The bacterial strains that will be investigated include: (i) endophytes, (ii) actinomycetes, and (iii) extremophiles from under-explored ecological niches in various parts of China. The criteria for strain selection are: (i) the uniqueness and richness of biodiversity, (ii) symbiosis with traditional Chinese medicinal plants, (iii) morphological and taxonomical distinctiveness, and (iv) bias towards actinomycetes. We will (i) optimize natural product production, (ii) prepare extracts and subject them to TLC and HPLC for chemical profiling and titer estimation followed by LC-MS and 1H NMR dereprelication for strain prioritization, (iii) isolate and determine the structures of the novel natural products, and (iv) provide them to MLSCN. Our hypotheses are: (i) bacteria that are morphologically distinctive and taxonomically new are likely to produce novel natural products, (ii) assay-blind and TLC- and HPLC-based chemical profilings allow the detection of all natural products, (iii) LC-MS and 1H NMR followup analyses permit rapid dereplication of the extracts, allowing prioritization of the strains for subsequent isolation and structural elucidation, and (iv) quantities of the natural products can be produced by scaleup microbial fermentation. The specific alms are: (i) to evaluate 600 strains, including approximately 200 endophytes, 300 actinomycetes, and 100 extremophiles, provided by Prof. Cheng-Lin Jiang of the Yunnan Institute of Microbiology, Yunnan University, China, by fermentation optimization, extract preparation, TLC- and HPLC-based chemical profiling and titer estimation, and LC-MS and 1H NMR analyses to identify 15 hit strains annually; (ii) to dereplicate the natural products in the 15 hit strains by natural product isolation and structural elucidation to identify one to five novel natural products per strain to yield a total of 15-75 compounds annually; and (iii) to scale up fermentation of the hit strains and to provide a 15-75-member library of the resultant novel natural products, each of which will be HPLC and NMR pure and in quantities weighing minimally 10 mg, to MLSCN annually. PUBLIC HEALTH RELEVANCE: Natural products have inspired ~75% of the drugs approved for human use and are evolutionarily optimized to serve as modulators of biologically important processes. This application seeks to discover natural products from new microorganisms, isolated from previously untapped and carefully selected ecological niches, for use as biochemical tools by which to understand fundamental and applied biology.

描述（由申请人提供）：我们建议从尚未开发的微生物中分离出新型天然产物，并将其提供给分子图书馆筛选中心网络（MLSCN）。将研究的细菌菌株包括：（i）内生菌，（ii）放线菌，和（iii）来自中国各地尚未开发的生态位的极端微生物。菌株选择的标准是：（i）生物多样性的独特性和丰富性，（ii）与传统中药植物的共生性，（iii）形态和分类学的独特性，以及（iv）偏向放线菌。我们将 (i) 优化天然产物的生产，(ii) 制备提取物并进行 TLC 和 HPLC 进行化学分析和滴度估计，然后进行 LC-MS 和 1H NMR 去重复以进行菌株优先级排序，(iii) 分离并确定其结构新颖的天然产物，以及 (iv) 将其提供给 MLSCN。我们的假设是：(i) 形态独特且分类学上新颖的细菌可能会产生新型天然产物，(ii) 盲法以及基于 TLC 和 HPLC 的化学分析可以检测所有天然产物，(iii) LC -MS 和 1H NMR 后续分析可以快速消除提取物的重复，从而可以对菌株进行优先排序以进行后续分离和结构阐明，并且 (iv) 可以生产大量的天然产物通过放大微生物发酵。具体任务是：（i）对中国云南大学云南微生物研究所蒋成林教授提供的600株菌株进行评价，包括约200种内生菌、300种放线菌和100种极端微生物，通过发酵优化、提取制备、基于 TLC 和 HPLC 的化学分析和滴度估计，以及 LC-MS 和 1H NMR 分析，以鉴定 15 种命中菌株每年； (ii) 通过天然产物分离和结构解析，对 15 个命中菌株中的天然产物进行去复制，以鉴定每个菌株 1 至 5 个新型天然产物，每年产生总共 15 至 75 种化合物； (iii) 扩大命中菌株的发酵规模，并每年向 MLSCN 提供由 15-75 个成员组成的新型天然产物文库，其中每种产物均为 HPLC 和 NMR 纯，且数量至少为 10 mg。 公共健康相关性：约 75% 的批准用于人类使用的药物受到天然产物的启发，并经过进化优化，可作为重要生物学过程的调节剂。该应用旨在从新微生物中发现天然产物，这些微生物是从以前未开发和精心挑选的生态位中分离出来的，用作了解基础生物学和应用生物学的生化工具。