Bryan, SamanthaCraig, SeanDanesh-Azari, Hamid-RezaAlHajji, Fatimah2024-12-162024https://hdl.handle.net/20.500.14154/74243To facilitate the synthesis of paclitaxel, a potent chemotherapy drug extensively used to treat a wide range of cancer types., taxadiene is an essential precursor that must be present. Because of the sluggish growth rate of the Pacific yew tree and the small amount of chemicals it produces, the conventional method of obtaining paclitaxel from the tree is not conducive to preserving the environment. To address this, researchers have begun investigating alternative chemical processes to produce paclitaxel. The purpose of this research was to design a genetic construct for the biosynthesis of taxadiene, insert the taxadiene synthase (TaSy) gene into the pL99 vector, and evaluate the antibiotic sensitivity of eight distinct Streptomyces species to ascertain their suitability as hosts for genetic material and taxadiene production using the organism's inherent capacity to produce secondary metabolites. In this work, the genes for taxadiene synthase (TaSy) and geranylgeranyl diphosphate synthase (GGPPS) were successfully introduced into some of these strains of Streptomyces. The technique started with successfully cloning a plasmid that included the required genes into E. coli., then conjugating this into Streptomyces bacteria. Streptomyces sp. SYN 562 successfully maintained the plasmid, which was evaluated using mass spectrometry, revealing a distinct peak associated with the molecular weight of taxadiene at 272 amu. Through this research, the capability of Streptomyces as an organism that is suitable for the synthesis of taxadiene in an environmentally friendly manner is brought to light. It serves as a basis for further research that will be conducted in the future to enhance the metabolic process and boost production on a larger scale for use in industrial applications. The findings provide support for the continuing attempts to create ways that are both more environmentally friendly and more cost-effective to produce paclitaxel, which is an essential component for the treatment of cancer.30enTaxadieneStreptomycesPaclitaxelgeranylgeranyl diphosphatebiosynthesisE. coli.Thesis