Heterologous Production of Eleutherobin Precursors

描述（由申请人提供）：萜类化合物是一类具有多种生物活性的天然化合物。由于萜类化合物的实际用途取决于提取或合成这些化合物的能力，因此只有最丰富或最容易合成的化合物才被商业化开发。这项小型企业技术转让第一阶段项目的长期目标是通过经济地生产稀缺的萜烯，促进新的抗癌化合物的临床试验和使用。刺红素和肌酸苷A和B是细胞毒性萜，其作用方式类似于非常成功的抗癌药物紫杉醇。然而，供应的限制阻碍了这些令人兴奋的化合物的发展。本I期研究将采用创新的方法鉴定、克隆和表达负责形成刺柳藿苷碳骨架的萜烯环化酶。通过检测环化酶的活性，我们将分离出单胞烷环化酶并对其进行部分测序。这些部分序列将用于设计PCR探针克隆全长环化酶基因。环化酶的序列将作为发现完整生物合成途径的起点。由于海洋动物产生的许多化合物是微生物共生体的产物，环化酶基因允许识别与我们感兴趣的化合物相关的生物合成基因簇。在过量生产萜烯前体的细菌中发现和表达已确定的生物合成途径将允许在大规模发酵中生产有价值的萜烯。这项与加州大学合作进行的研究将有助于扩大萜烯环化酶的结构、功能和进化领域，并将为研究生和本科生提供教育机会。这项技术可以开发出前景光明的新型萜烯类药物，在对自然资源的破坏和合成化学废水的释放方面，对消费者和环境的成本都较低。

DESCRIPTION (provided by applicant): Terpenoids are a diverse class of natural compounds with equally diverse and potent bioactivities. Because the practical use of terpenoids hinges on the ability to extract or synthesize these compounds, only the most abundant or easily synthesized compounds have been commercially developed. The long-term goal of this Small Business Technology Transfer Phase I project is to facilitate the clinical testing and use of new cancer compounds through the economical production of otherwise scarce terpenes. Eleutherobin and sarcodictyin A and B are cytotoxic terpenes that act in a manner analogous to very successful anti-cancer drug Taxol. Supply limitations, however, has precluded the development of these exciting compounds. This Phase I research will use innovative methods to identify, clone and express the terpene cyclase responsible for the formation of the eleutherobin carbon backbone. By detecting the activity of the cyclase, we will isolate and partially sequence the eunicellane cyclase. These partial sequences will then be used to design PCR probes to clone the full length cyclase gene. The sequence of the cyclase will act as a starting point in the discovery of the full biosynthetic pathway. As many compound produced by marine animals are the product of microbial symbionts, the cyclase gene allow the identification of biosynthetic gene clusters associated with our compound of interest. The discovery and expression of the identified biosynthetic pathway in bacteria that over-produce terpene precursors will allow the production of valuable terpenes in large scale fermentations. This research, conducted in collaboration with the University of California, will contribute to the expanding field of terpene cyclase structure, function and evolution and will provide educational opportunities for graduate and under-graduate students. This technology could result in the development of promising new terpene-based drugs produced at a lower cost to consumers and to the environment, in terms of destruction of natural resources and the release of synthetic chemical effluents.