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出新的天然产物，并将其提供给分子文库筛选中心网络（MLSCN）。将研究的菌株包括：(i)内生菌，（ii）放线菌，（iii）来自中国各地未开发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）与传统药用植物的共生关系，（iii）形态和分类上的独特性，以及（iv）对放线菌的偏好。我们将(i)优化天然产物的生产，（ii）制备提取物并对其进行TLC和HPLC化学分析和滴度估计，然后进行LC-MS和1H NMR去预处理以确定菌株的优先级，（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)形态独特且分类新颖的细菌可能产生新的天然产物，（ii）基于分析盲和TLC和hplc的化学分析允许检测所有天然产物，（iii） LC-MS和1H NMR后续分析允许快速分离提取物，允许优先菌株进行后续分离和结构阐明，以及（iv）可以通过大规模微生物发酵生产大量天然产物。(1)利用云南大学云南微生物研究所蒋成林教授提供的600株菌株，包括内生菌约200株、放线菌约300株、极端微生物约100株，通过发酵优化、提取液制备、TLC和hplc化学分析和滴度估计、LC-MS和1H NMR分析，每年鉴定出15株hit菌株；（ii）通过天然产物分离和结构解析，在15个命中菌株中重复天然产物，每个菌株鉴定1至5个新的天然产物，每年产生15-75个化合物；（iii）扩大成功菌株的发酵规模，并提供15-75个成员的新天然产物库，每个产物将是HPLC和NMR纯的，数量不超过10毫克，每年达到MLSCN。

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.