Development of a Bacterial Host for Natural Product Discovery and Production

用于天然产物发现和生产的细菌宿主的开发

• 批准号: 10439880
• 负责人: Alessandra S Eustaquio
• 金额: $ 36.96万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-20 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10439880
• 关键词: Address; Anabolism; Autologous; Bacteria; Bacterial Genome; Bacterial Model; Biological Assay; Breath Tests; Burkholderia; Collection; Communities; Coupled; DNA; Data; Development; Escherichia coli; Gene Cluster; Genes; Genome; Goals; Grant; Growth; Health; Human; Knowledge; Lasso; Libraries; Medicine; Mining; Modeling; Natural Products; Organism; Pathway interactions; Peptide Biosynthesis; Peptides; Play; Privatization; Process; Production; Proteobacteria; Regulation; Regulator Genes; Reporter; Role; Source; Structural Genes; Taxonomy; Technology; Testing; Transcription Coactivator; base; bioactive natural products; drug discovery; genome sequencing; interest; mass cell culture; novel; peptide B; promoter; resistance gene; synthetic biology; tool; transcriptome sequencing

Project Summary Molecules isolated from living systems have consistently served as leads in drug discovery. However, two of the main challenges of drug discovery from natural products are the high rate of rediscovery of known compounds using traditional technologies and low yields. Recent advances in genome sequencing and synthetic biology have spurred a renewed interest in natural product discovery in the private and academic sectors. For instance, genome mining of underexplored taxa increases the chances of novel compound discovery. Moreover, robust host organisms can facilitate discovery efforts by overcoming the common low-yield hurdle. Bacteria belonging to the Burkholderiales order of -Proteobacteria are an emerging source of natural products. We have previously achieved high yield production of autologous polyketide-nonribosomal peptide spliceostatins in a Burkholderia species. We propose to understand, develop, and apply this strain as a host to discover and produce natural products from Burkholderiales and potentially other -Proteobacteria. Although Escherichia coli has long been used as a model bacterial host, the synthetic biology community is moving away from the idea that “one host fits all” to instead have hosts tailored to the source of biosynthetic gene clusters. We hypothesize that the Burkholderia sp. in question can be used as a host to streamline the discovery process by producing heterologous natural products in high yields. Our preliminary data supports this hypothesis as heterologous expression of a model gene cluster encoding the lasso peptide capistruin in this strain led to capistruin production in yields that are at least 65-fold, and up to 580-fold higher than with E. coli. In Aim 1 we propose to test/understand the host by a) testing the breath of the host in terms of source genera within the Burkholderiales and other -Proteobacteria while discovering lasso peptides; and b) investigating the regulation of autologous spliceostatin biosynthesis with the ultimate goal of deriving regulatory parts for heterologous pathway construction. In Aim 2, we will develop the host as a tool for natural product discovery by discovering and characterizing a promoter library and by generating a minimized genome. In Aim 3 we will apply the host by discovering natural products from a newly built Burkholderiales environmental collection. This project is expected to provide tools that will streamline and accelerate natural product discovery from a promising yet underexplored source. The tools and knowledge obtained will be applied to discover novel, bioactive natural products via genome mining of a Burkholderiales strain collection.

项目摘要 从生活系统中分离出来的分子一直是药物发现的铅。但是，两个 从天然产品发现药物发现的主要挑战是已知化合物的高分速率 使用传统技术和低收率。基因组测序和合成生物学的最新进展 对私人和学术领域的自然产品发现产生了新的兴趣。例如， 未经置换的分类单元的基因组开采增加了新型复合发现的机会。而且，健壮 寄主生物可以通过克服普通的低收益障碍来促进发现工作。细菌属于 对于burkholderiales的burkholderiales droteobacteria命令是新兴天然产品的来源。我们以前有 在Burkholderia中获得了自体聚酮化 - 非核体肽剪接素的高收益产生 物种。我们建议理解，发展和运用这种压力作为宿主，以发现和产生自然 来自Burkholderiales和其他可能的totototobacteria的产品。虽然大肠杆菌长期以来一直是 合成生物学社区用作模型细菌宿主，远离了“一个宿主适合的观念 取而代之 Burkholderia sp。中间可以用作主机来通过生产来简化发现过程的主机 高产的异源天然产物。我们的初步数据支持这一假设是异源的 在这种菌株中编码套索胡椒蛋白的模型基因簇的表达导致capistruin的产生 在至少65倍的产率中，比大肠杆菌高出580倍。在目标1中，我们建议 通过a）测试/理解主机。 在发现套索肽的同时，其他蛋白菌病； b）调查自体的调节 剪接素生物合成的最终目标是推导异源途径的调节部分 建造。在AIM 2中，我们将通过发现和 表征启动子库并产生最小化的基因组。在AIM 3中，我们将申请主机 从新建的Burkholderiales环境收藏中发现天然产品。期望这个项目 提供可以从诺言中简化和加速自然产品发现的工具 来源。获得的工具和知识将用于发现通过 Burkholderiales菌株收集的基因组开采。