Figure 3

Effects of pH (A) and temperature (B) on the lactate production through the in vitro synthetic pathway. Enzymes (0.1 U ml⁻¹ each) were incubated in a mixture of 10 mM glycerol, 1 mM NAD⁺, 0.2 mM NADH, 0.2 mM ATP, 0.2 mM ADP, 0.02 mM FAD, 0.5 mM FBP, 1 mM G1P, 5 mM MgCl₂, 0.5 mM MnCl₂, and an appropriate buffer. Reaction was performed at indicated pH and temperature for 30 min and then terminated by removing the enzymes with ultrafiltration. Fifty millimolar of MES-NaOH (pH 6) and HEPES-NaOH (pH 7 and 8) were used to adjust pH.