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

本研究的目的是量化色氨酸过量产生对长春花毛状根组织培养中中心碳代谢和吲哚生物碱途径的代谢影响。本研究主要有四个方面的目标：(1)建立过表达甲氨酸合酶和色氨酸脱羧酶的转基因玫瑰毛状根无性系；(2)上述菌株在生长、营养水平、氨基酸库和吲哚生物碱积累方面的特征；(3)细胞内通量的数学分析；(4)使用新的体外13c核磁共振技术测量细胞内通量。

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.