Figure 4

Time course hydrolysis and TG catalyzed by EcChi1 with DP3, DP4. The reaction mixture was incubated with 200 nM of EcChi1 and 1 mM of DP3/DP4 for different time periods from 0 to 720 min at 40 °C. Products were analyzed by binary gradient HPLC. (A and B) HPLC profiles of reaction products from DP3 (A) and DP4 (B) substrates. The topmost profile shows a standard mixture of CHOS ranging from DP1 to DP6 (A and B). The other profiles show the reaction products from DP3 and DP4 substrates at the indicated incubation times. The inset shows a magnified view of the low-peak-area products. Control represents the substrate without EcChi1. Overview of concentrations of CHOS products generated during reaction time courses with DP3 (C) and DP4 (D) substrates. Products were quantified from respective peak areas using standard calibration curves of CHOS ranging from DP1 to DP6.