Natural Products from Under-explored Microorganisms

描述（由申请人提供）：我们建议从未开发的微生物中分离新型天然产物，并将其提供给分子库筛选中心网络（MLSCN）。将被调查的细菌菌株包括：（i）内生菌，（ii）放线菌，和（iii）极端微生物，从中国各地未开发的生态位。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关系，（iii）形态和分类的独特性，以及（iv）对放线菌的偏好。我们将（i）优化天然产物的生产，（ii）制备提取物，并将其进行TLC和HPLC，以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）分离并确定新型天然产物的结构，以及（iv）将其提供给MLSCN。我们的假设是：（i）形态上独特且分类学上新的细菌可能产生新的天然产物，（ii）基于分析盲法以及TLC和HPLC的化学分析允许检测所有天然产物，（iii）LC-MS和1H NMR后续分析允许提取物的快速去复制，允许菌株的优先级排序用于随后的分离和结构阐明，和（iv）可以通过放大微生物发酵生产大量的天然产物。具体的施舍是：（i）通过发酵优化、提取物制备、基于TLC和HPLC的化学分析和滴度估计以及LC-MS和1H NMR分析，评估中国云南大学云南微生物研究所的蒋成林教授提供的600种菌株，包括约200种内生菌、300种放线菌和100种极端微生物，以每年鉴定15种命中菌株;（ii）通过天然产物分离和结构解析，去复制15个命中菌株中的天然产物，以鉴定每个菌株1至5种新的天然产物，从而每年产生总共15-75种化合物;和（iii）扩大命中菌株的发酵并提供所得新天然产物的15-75个成员的文库，每种样品均为HPLC和NMR纯品，且重量至少为10 mg，每年供应至MLSCN。 公共卫生相关性：天然产物激发了约75%的批准用于人类使用的药物，并且经过进化优化，可作为生物学重要过程的调节剂。本申请旨在发现来自新微生物的天然产物，这些新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，可用作理解基础和应用生物学的生物化学工具。

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.