MANIPULATION OF T BREVIFOLIA PHYSIOLOGY IN TISSUE CULTUR

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This project is directed towards a broad-based understanding of the manipulation of tissue cultures of Taxus brevifolia and related species for the production of taxol or taxol-like compounds which could be easily converted to taxol. Taxol is very promising anti-cancer agent which is in extremely short supply. A better understanding of how to manipulate cellular synthesis of taxol in tissue culture will provide the basis for efficient 'factory-like' production of taxol and thereby alleviate potential supply problems. The specific component goals of the project includes enabling methods such as development of rapid assay techniques for taxol; establishment of new cultures, particularly root cultures; evaluation of taxol self-inhibition and possible mechanisms of inhibition; and, characterization of taxol partition coefficients on a variety of resins and into organic solvents. With these techniques we will probe the effects of gas phase composition, elicitors, in situ product removal, and cell immobilization on cellular growth rates, taxol production rates, ratio of extracellular to intracellular taxol, and production of taxol-related compounds. Further, we will examine the effects of liquid shear in stirred reactors on cell physiology. Also, novel small-scale devices will be used to test the intrinsic capacity of root cultures for growth and product formation.