Fig. 5: Metabolite concentrations at equilibrium differ between overoxidizing Mdh2 CT4-1 Cn and non-overoxidizing Mdh Bm MGA3. | Nature Communications

Fig. 5: Metabolite concentrations at equilibrium differ between overoxidizing Mdh2 CT4-1 Cn and non-overoxidizing Mdh Bm MGA3.

From: Identification of overoxidizing and non-overoxidizing NAD-dependent methanol dehydrogenases and implications for synthetic methylotrophy

Fig. 5: Metabolite concentrations at equilibrium differ between overoxidizing Mdh2 CT4-1 Cn and non-overoxidizing Mdh Bm MGA3.

a In vitro assays of Mdh2 CT4-1 Cn (yellow), Mdh Bm MGA3 (olive green), and Mdh Bm MGA3 + Act (purple) monitored until equilibrium was reached. Reactions contained 25 μM Mdh2 CT4-1 Cn or 25 μM Mdh Bm MGA3, 2.3 μM Act if indicated, 5 mM NAD+ and were started by the addition of 500 mM methanol. The mean of the data is shown as solid line and the shaded regions represent the range of the 90% confidence intervals. b Methanol oxidation to formaldehyde and its subsequent oxidation to formate. ΔrG’0 values were calculated by Equilibrator (pH = 7.4, pMg = 2.3, ionic strength = 0.15 M)96,101. c Equilibrium concentrations of formaldehyde (blue), formate (red), and NADH (yellow). Reactions contained 25 μM Mdh2 CT4-1 Cn or 25 μM Mdh Bm MGA3, 2.3 μM Act if indicated, 5 mM NAD+ and were started by the addition of 500 mM methanol. Experiments were performed in triplicates and data are represented as mean ± the standard deviation. Source data are provided as a Source Data file.

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