Project 463073

Taxol is one of the best anticancer drugs ever developed, used against ovarian, breast, lung and other cancers. Its effectiveness combined with anticipated further therapeutic applications has generated huge demand for the drug, which outpace current supply. Yet, an inexpensive taxol production method is lacking, raising the cost of treatment for one patient to $7000 for 6 cycles of chemotherapy. Taxol extraction from natural sources demands destruction of two adult yew trees to obtain 1 g of compound. Chemical synthesis is very complicated and inefficient, hence not suitable for industrial scale. A sustainable and economical solution is production of taxol in simple organisms, such as yeast. For this purpose, the entire enzymatic route of taxol formation that naturally occurs in plants must be transplanted in yeast. However, not all enzymes involved in taxol biosynthesis are known and how they work together along the 19 steps is yet unclear. We will use synthetic biology, computational science and analytical and organic chemistry to develop high throughput approaches for engineering yeast cells to produce complex taxol intermediate compounds, which could be further chemically modified to taxol or other promising analogs. This will be achieved by discovery and clarification of yet unknown or ambiguous reactions. Subsequently, we will introduce in the system pathway-unrelated enzymes to enable new modifications of the taxol natural skeleton and synthesis of novel compounds. Following optimization of precursors, pathway balancing and establishment of optimal culture parameters, we expect to obtain increased yields of intermediate compounds of interest for bioactivity assays and future applications and benefit cancer patients.