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Figure 1.

Flowchart of dFBA to decipher the dynamic metabolism of S. oneidensis MR-1.

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Figure 2.

Monod model for growth kinetics.

The green dots are the measurements, and the blue lines are the simulated growth by the empirical Monod model.

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Table 1.

Parameters estimated in the empirical Monod model.

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Figure 3.

Prediction of growth rates (h−1).

Blue ○: growth rate determined by the Monod model. Red ○: dFBA prediction using the objective function (maximization of growth rate). Green □: dFBA prediction using dual-objective functions (maximization of growth rate and minimization of overall flux). Yellow ◊: the weight of the dual-objective functions that predicted the measured growth rates. Note: the summation of the square of fluxes (∑v2) was a very large number (total 774 fluxes), so the magnitude of weight w was small.

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Figure 4.

Dynamic flux distributions (unit: mmol/g DCW/h) in central metabolic pathways.

The yellow filled cycles are intracellular metabolites; the blue filled cycles are substrates and extracellular metabolites (LAC: extracellular lactate, PYR: extracellular pyruvate, ACT: extracellular acetate); the dashed lines indicate inactive pathways; the green filled boxes are reactions listed in iSO783. All the abbreviations refer to iSO783 [7].

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Figure 5.

Experimentally observed and simulated isotopomer labeling patterns [M-57]+ in proteinogenic amino acids.

The standard error for GC-MS measurement was below 0.02. A1: dynamic isotopomer simulation for glutamate from dFBA without considering reaction reversibility (dFBA w/o reversibility). A2: dynamic isotopomer simulation for glutamate from dFBA considering reaction reversibility (dFBA w/ reversibility). Bar plot: comparison of experimentally observed isotopomer labeling to simulated isotopomer labeling patterns of glutamate (A1: without considering reaction reversibility; A2: considering reaction reversibility). B: The model fitting of the isotopomer labeling data of five key amino acids (Ala, Gly, Ser, Asp, and Glu) at t = 24 and 30 h.

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Table 2.

Exchange coefficients for key metabolic pathways of MR-1.

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Table 2 Expand