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利用动力学代谢模型分析降低细胞质三磷酸甘油醛异构酶对马铃薯毛状根细胞的影响。

Analyzing the effect of decreasing cytosolic triosephosphate isomerase on Solanum tuberosum hairy root cells using a kinetic-metabolic model.

机构信息

Canada Research Chair in Applied Metabolic Engineering, Bio-P² Research Unit, Department of Chemical Engineering, École Polytechnique de Montreal, Montreal, Quebec, Canada.

出版信息

Biotechnol Bioeng. 2013 Mar;110(3):924-35. doi: 10.1002/bit.24747. Epub 2012 Oct 18.

Abstract

A kinetic-metabolic model of Solanum tuberosum hairy roots is presented in the interest of understanding the effect on the plant cell metabolism of a 90% decrease in cytosolic triosephosphate isomerase (cTPI, EC 5.3.1.1) expression by antisense RNA. The model considers major metabolic pathways including glycolysis, pentose phosphate pathway, and TCA cycle, as well as anabolic reactions leading to lipids, nucleic acids, amino acids, and structural hexoses synthesis. Measurements were taken from shake flask cultures for six extracellular nutrients (sucrose, fructose, glucose, ammonia, nitrate, and inorganic phosphate) and 15 intracellular compounds including sugar phosphates (G6P, F6P, R5P, E4P) and organic acids (PYR, aKG, SUCC, FUM, MAL) and the six nutrients. From model simulations and experimental data it can be noted that plant cell metabolism redistributes metabolic fluxes to compensate for the cTPI decrease, leading to modifications in metabolites levels. Antisense roots showed increased exchanges between the pentose phosphate pathway and the glycolysis, an increased oxygen uptake and growth rate.

摘要

为了了解反义 RNA 对细胞质三磷酸甘油醛异构酶 (cTPI,EC 5.3.1.1) 表达降低 90%对植物细胞代谢的影响,提出了马铃薯毛状根的动力学-代谢模型。该模型考虑了主要代谢途径,包括糖酵解、戊糖磷酸途径和 TCA 循环,以及导致脂质、核酸、氨基酸和结构己糖合成的合成反应。从摇瓶培养中测量了六种细胞外营养物质(蔗糖、果糖、葡萄糖、氨、硝酸盐和无机磷酸盐)和 15 种细胞内化合物,包括糖磷酸(G6P、F6P、R5P、E4P)和有机酸(PYR、aKG、SUCC、FUM、MAL)和六种营养素。从模型模拟和实验数据可以看出,植物细胞代谢重新分配代谢通量以补偿 cTPI 的减少,导致代谢物水平的变化。反义根显示戊糖磷酸途径和糖酵解之间的交换增加,耗氧量和生长速率增加。

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