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A method is presented to identify flux controlling reactions in metabolic networks using experimentally determined flux distributions.
We identify flux distributions for which the calculated labeling patterns agree well with the measurements alluding to the accuracy of the network reconstruction.
Recently, OptForce [ 21] was used to identify flux manipulation leading to targeted overproductions.
Besides, FBA uses an objective function to identify flux distributions that maximize (or minimize) the physiologically relevant predicted solution.
These constraints yield a solution space of possible flux values, and FBA uses an objective function to identify flux distributions that maximize (or minimize) the physiologically relevant predicted solution.
We then used a combination of metabolic flux predictions and mutational analysis to identify flux redistribution patterns utilized in the Δ pntAB mutant to compensate for the loss of this enzyme.
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However, using NMR data to identify fluxes can be problematic because a non-linear optimization problem can result that is highly non-convex due to the bi-linear and tri-linear terms present in the isotopomer balance equations.
The set of simultaneous linear and non-linear differential equations that constitute deterministic models can be investigated using a form of sensitivity analysis (developed in the 1960s by chemical engineers [ 19], and now a part of metabolic control analysis [ 11]) to help identify flux-controlling steps (enzymes) that then become the target for genetic manipulations of the organism [ 5].
Based on the identified flux distributions or linear combinations, the user can simulate the NMR and GC/MS spectra of selected signal molecules.
The identified flux ranges of solventogenic nutrients (glucose, acetate, butyrate, and carbon dioxide) were subsequently calculated by fixing both the growth rate and restricting the acids/solvents to the FVA calculated maximum and minimum values.
In addition, we also identified flux-sum intensification targets, such as glyoxylate, 2-phosphoglycerate and 3-phosphoglycerate, which were also reported as effective targets for increasing succinate production (see Additional file 2).
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