Natural Products from Under-explored Microorganisms

描述(由申请人提供)：我们建议从未被开发的微生物中分离出新的天然产物，并将它们提供给分子图书馆筛选中心网络(MLSCN)。将进行调查的细菌菌株包括：(I)内生菌，(Ii)放线菌，(Iii)来自中国各地未被开发的生态位的极端细菌。选择菌株的标准是：(I)生物多样性的独特性和丰富性；(Ii)与传统药用植物的共生；(Iii)形态和分类上的独特性；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，并对其进行TLC和HPLC化学图谱和效价评估，然后进行LC-MS和1H核磁共振去复制以确定菌株的优先顺序，(Iii)分离和确定新天然产物的结构，(Iv)将它们提供给MLSCN。我们的假设是：(I)在形态上独特的和分类上新的细菌可能产生新的天然产物，(Ii)基于分析盲和基于薄层和高效液相的化学图谱允许检测所有天然产物，(Iii)LC-MS和1HNMR后续分析允许快速去复制提取物，允许优先排序菌株用于随后的分离和结构鉴定，以及(Iv)大量的天然产物可以通过扩大微生物发酵产生。具体措施是：(1)对云南大学云南微生物研究所蒋成林教授、中国教授提供的600株细菌进行鉴定，包括近200株内生菌、300株放线菌和100株极端细菌，通过发酵优化、提取物制备、基于薄层和高效液相的化学图谱和效价测定以及LC-MS和1H核磁共振分析，每年鉴定15株热门菌株；(Ii)通过天然产物分离和结构鉴定，对15株热门菌株中的天然产物进行去复制，每个菌株鉴定1~5种新的天然产物，每年总共产生15~75种化合物；以及(Iii)扩大Hit菌株的发酵，并每年向MLSCN提供15-75个成员的新天然产物库，其中每一个都将是高纯度的高效液相和核磁共振，质量至少为10毫克。 与公共健康相关：天然产品激发了约75%被批准用于人类的药物的灵感，并经过进化优化，作为生物重要过程的调节器。这项应用寻求从新的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，用作了解基础和应用生物学的生化工具。

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.