Fig. 1

HPLC elution profiles of the selected reaction mixtures. Enzymatic reaction was conducted in the following conditions: 25 mM G2, 100 mM αGlc1-P (αGlc1-P/G2 ratio of 4.0), 1 U/mL CcCdP in 50 mM MES buffer (pH 7.0), at 45 °C with reaction time of a, 5 min; b, 20 min; c, 40 min; d, 90 min; and e, 360 min. Analysis was done on a Hitachi LaChrom HPLC system equipped with a Luna 5 µm NH₂ column (100 Å, 250 × 4.6 mm) operated at 40 °C. Acetonitrile–water (70:30, v/v) was used as eluent at a flow rate of 1.5 mL/min. Refractive index detection was used. Axis scale of detector response (y) was the same for a-e, and d’ was the profile of d shown with smaller scale.