A sequenced-based approach for improved small molecule discovery

描述（由申请人提供）：DNA测序技术的最新进展和对天然产物生物合成的更好理解为改进微生物天然产物的发现过程提供了新的机会。本研究的目的是建立一系列的方法，通过该方法可以快速评估菌株的天然产物的生物合成，通过分析PCR产生的或基因组序列数据。这些方法将使用属于Salinispora属的海洋细菌的模型组开发，然后应用于大量不同的海洋放线菌，目的是发现结构多样的新化学实体，这些实体将提供给NIH分子图书馆小分子储存库（MLSMR）。这些方法包括一个初始的、快速的分子“指纹”筛选，从中可以比较各个菌株的遗传潜力。然后，基于序列的方法将被应用于解释生物合成的丰富性和新奇的菌株与有前途的指纹。这些方法将使预测菌株产生的代谢物是否是新的以及在特定结构类别中可能产生多少不同的化合物成为可能。一旦确定具有最大遗传潜力的菌株，将进行详细的化学研究。这种方法代表了一个显着的改进，在传统的范例中，大量的生物合成未知的菌株在有限数量的条件下进行筛选。它将大大减少以前发现的化合物的分离，这是一个长期困扰微生物天然产物研究的问题。这种方法提供了一种不依赖于培养的、基因组水平的次级代谢物生物合成评估，而不是更传统的方法，该方法仅检测在有限的培养条件下产生的那些代谢物。开发的方法将广泛适用于科学界，包括创建一个精心策划的序列数据库，该数据库可以随时下载并用于评估次级代谢产物产生途径的基因组序列数据。这项研究有可能大大提高发现新化学实体并用于生物医学研究的速度。 公共卫生相关性：该提案中提出的研究提供了一种方法，通过这种方法，DNA序列数据可以用于显着改善从微生物中发现天然产物的过程。它将产生大量新的化学多样性，可用于药物发现研究和研究基本的生物过程。所开发的方法将广泛提供给研究界，从而对药物发现和基础生物医学研究产生广泛影响。

DESCRIPTION (provided by applicant): Recent advances in DNA sequencing technologies and a better understanding of natural product biosynthesis provide newfound opportunities to improve the process by which microbial natural products are discovered. The objectives of this research are to establish a series of methodologies by which strains can be quickly assessed for natural product biosynthesis through the analysis of PCR-generated or genome sequence data. The methods will be developed using a model group of marine bacteria belonging to the genus Salinispora and then applied to a large and diverse collection of marine actinomycetes with the aim of discovering structurally diverse, new chemical entities, which will be provided to the NIH Molecular Libraries Small Molecule Repository (MLSMR). The methods include an initial, rapid molecular "fingerprinting" screen, from which the genetic potential of individual strains can be compared. Sequence-based approaches will then be applied to interpret the biosynthetic richness and novelty of strains with promising fingerprints. These methods will make it possible to predict if the metabolites produced by a strain will be new and how many different compounds in a particular structural class may be produced. Once strains with the greatest genetic potential are identified, detailed chemical studies will be performed. This approach represents a dramatic improvement over traditional paradigms in which large numbers of biosynthetically unknown strains are screened in a limited number of conditions. It will dramatically reduce the isolation of previously discovered compounds, a problem that has long plagued microbial natural product research. This approach provides a culture-independent, genome-level assessment of secondary metabolite biosynthesis as opposed to more traditional methods, which detect only those metabolites produced under a limited set of culture conditions. The methods developed will be broadly applicable to the scientific community and include the creation of a curated sequence database that can be readily downloaded and used to assess genome sequence data for pathways involved in secondary metabolite production. This research has the potential to dramatically increase the rates with which new chemical entities are discovered and made available for biomedical research. PUBLIC HEALTH RELEVANCE: The research presented in this proposal provides a method by which DNA sequence data can be used to dramatically improve the process by which natural products are discovered from microorganisms. It will generate considerable new chemical diversity that can be used for drug discovery research and to study basic biological processes. The methods developed will be made widely available to the research community and thereby have a broad impact on drug discovery and basic biomedical research.