Figure 7.

GDP‐mannose concentration over time for cascade reaction starting from mannose, ADP, GDP, and PolyP₁₄. Enzyme concentrations were His₆‐Glk 0.5 mg/ml, His₆‐1D‐Ppk2 0.05 mg/ml, ManB‐His6/ManC 0.3 mg/ml, and PmPpA‐His₆ 0.1 mg/ml. Initial substrate concentrations are ADP 0.8 mM, GDP 0.8 mM, mannose 6 mM, and PolyP₁₄ 4 mM; reaction conditions were pH 7.5, 30°C and 10 mM MgCl₂ unless stated otherwise. The error bars of the standard reaction represent the standard deviation of the mean calculated from four replicates. (a) Experimental data (dots) and modeled data with k_4,opt (lines) for various initial concentrations; (b) GDP‐mannose concentration for reactions at pH values 7.0, 7.5, and 8.0; (c) GDPmannose concentration for reactions at 25, 30, and 35°C; (d) GDP‐mannose concentrations for MgCl₂ concentrations of 5, 10, and 20 mM. The fits depicted are fits to the entire data set of multi‐enzyme catalyzed reactions