Quantifying and Directing Metabolic Flux: Application to Tryptophan Overproduction and Indole Alkaloid Accumulation in Plant Tissues

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The objective of this project is to quantify the metabolic consequences of tryptophan overproduction upon central carbon metabolism and indole alkaloid pathways in Catharanthus roseus hairy root tissue cultures. This research has four objectives which include: (1) the generation of transgenic C. roseus hairy root clones that overexpress anthranilate synthase and tryptophan decarboxylase; (2) the characterization of the above strains in terms of growth, nutrient levels, amino acid pools, and indole alkaloid accumulation; (3) the mathematical analysis of intracellular fluxes; and (4) the measurement of intracellular fluxes using new in vitro 13 C NMR techniques.