Symbiont Models for Natural Product Pathway Manipulation

天然产物途径操纵的共生模型

• 批准号: 7032744
• 负责人: Eric W Schmidt
• 金额: $ 31.29万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-06 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7032744
• 关键词: Cyanophyta; Tunicata; aquaculture; bacterial proteins; bacteriocin; bioengineering /biomedical engineering; biological products; biological signal transduction; chemical synthesis; cyclic peptides; drug discovery /isolation; functional /structural genomics; fusion gene; gene expression; high performance liquid chromatography; high throughput technology; molecular cloning; nuclear magnetic resonance spectroscopy; peptide library; protein signal sequence; protein structure function; recombinant DNA; recombinant proteins; site directed mutagenesis; symbiosis; transfection /expression vector

DESCRIPTION (provided by applicant): Marine invertebrates, such as sponges, tunicates, and gorgonians, produce a stunning diversity of secondary metabolites, many of which show promise as pharmaceutical agents. However, supplying marine natural products for clinical development has been a major hurdle, since the organisms are often rare or difficult to collect. Recombinant DNA technology offers a possible strategy to circumvent the supply problem by expression of biosynthetic genes in bacterial culture, but there are many problems that have prevented the widespread use of this approach. The large bacterial diversity found within marine invertebrates and the difficulty of whole pathway expression greatly hinder the usefulness of the approach. In addition, it is often difficult to engineer pathways to produce new natural products. We recently reported the first example of rational whole-pathway identification and heterologous expression for a marine natural product from an animal-bacterium symbiosis. A pathway encoding formation of patellamides A and C was identified in Prochloron, the obligate symbiont of an ascidian, and expressed in Escherichia coli. In this project, we will capitalize on this early success by cloning and engineering biomedically important natural product pathways from a variety of symbiotic associations in the marine environment. By further exploring this model symbiotic system, we will establish and validate methodology for the general cloning of symbiotic products, develop a supply of drugs that are currently in clinical or preclinical trials, and discover new drugs via rational pathway engineering. This study will thus provide practical solutions to problems in drug discovery and development with marine natural products.

描述（由申请人提供）：海洋无脊椎动物，例如海绵，上衣和戈贡人，产生了令人惊叹的次级代谢物的令人惊叹的多样性，其中许多是有望作为药物剂的希望。但是，为临床开发提供海洋天然产品一直是一个重大障碍，因为这些生物通常很少或难以收集。重组DNA技术提供了一种可能通过在细菌培养中表达生物合成基因来避免供应问题的策略，但是有许多问题阻止了这种方法的广泛使用。在海洋无脊椎动物中发现的大细菌多样性以及整个途径表达的难度极大地阻碍了该方法的有用性。此外，要设计出生产新天然产品的途径通常很难。我们最近报道了来自动物 - 细菌共生的海洋天然产物的合理全途径鉴定和异源表达的第一个例子。在prochloron（甲状腺素的强制性共生体）中鉴定了patellamides a和c的编码途径，并在大肠杆菌中表达。在这个项目中，我们将通过从海洋环境中的各种共生协会中克隆和工程进行生物医学重要的天然产品途径来利用这一早期成功。通过进一步探索这种模型共生系统，我们将建立和验证共生产品的一般克隆的方法，开发目前正在临床或临床前试验中的药物供应，并通过有理途径工程发现新药。因此，这项研究将为使用海洋天然产品在药物发现和发育中的问题提供实际解决方案。