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

本项目的目的是量化色氨酸过量生产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。本研究有四个目标：(1)建立高表达邻氨基苯甲酸合成酶和色氨酸脱羧酶的转基因玫瑰毛状根克隆；(2)上述菌株的生长、营养水平、氨基酸库和吲哚生物碱积累的特征；(3)细胞内通量的数学分析；(4)利用新的体外~(13)C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.