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.
Thesaurus Terms: alkaloid, antineoplastic, diterpene, drug design /synthesis /production drug discovery /isolation, enzyme biosynthesis, lyase Escherichia coli, coral, liquid chromatography mass spectrometry, molecular cloning, polymerase chain reaction
