Symbiont Models for Natural Product Pathway Manipulation

海洋无脊椎动物，如海绵，被囊动物和柳珊瑚，产生了惊人的多样性， 次级代谢物，其中许多显示出作为药物制剂的前景。然而，供应 海洋天然产品用于临床开发一直是一个主要障碍，因为生物体通常 罕见或难以收集。重组DNA技术提供了一种可能的策略，以规避 通过在细菌培养物中表达生物合成基因来提供问题，但是存在许多问题， 阻碍了这种方法的广泛使用。海洋中发现的大量细菌多样性 无脊椎动物和整个途径表达的困难极大地阻碍了 approach.此外，通常很难设计生产新天然产品的途径。 我们最近报道了第一个合理的全途径鉴定和异源的例子， 从动物-细菌共生体中表达海洋天然产物。编码途径 在Prochloron（一种海鞘的专性共生体）中发现了patellamides A和C的形成， 在大肠杆菌中表达。在这个项目中，我们将利用这一早期的成功， 工程生物医学重要的天然产物途径，从各种共生协会， 海洋环境。 通过进一步探索这个模型共生系统，我们将建立和验证方法， 共生产品的一般克隆，开发目前处于临床或临床前的药物供应 试验，并通过合理的途径工程发现新药。这项研究将提供实用的 解决海洋天然产物药物发现和开发中的问题。

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.