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

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

Biotechnology and bioengineering (2012-10-12)
Alexandre Valancin, Balasubrahmanyan Srinivasan, Jean Rivoal, Mario Jolicoeur
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triosephosphate Isomerase from baker′s yeast (S. cerevisiae), Type I, ammonium sulfate suspension, ~10,000 units/mg protein
Sigma-Aldrich
Triosephosphate Isomerase from rabbit muscle, Type X, lyophilized powder, ≥3,500 units/mg protein
Sigma-Aldrich
Triosephosphate Isomerase from rabbit muscle, Type III-S, ammonium sulfate suspension, ≥4,000 units/mg protein