Figure 5.

The effect of domain proximity on the efficiency of Jd1381. The panels show solubilization of GlcNAc (A) and oxidized products (B) during incubation of 10 g/liter α-chitin with 1 μm of Jd1381 with AscA (green circles) or JdCBM5-Chi18 (brown circles) or JdChi18 (white circles) or JdLPMO10 (orange circles) or JdLPMO10-CBM5 (black circles) or combinations of 1 μm JdLPMO10 and 1 μm JdCBM5-Chi18 (light blue circles) or 1 μm JdLPMO10-CBM5 and 1 μm JdChi18 (pink circles) in 20 mm BisTris (pH 6.0). The red circles show product formation in a reaction with Jd1381 without added AscA. Prior to use, all enzymes were copper-saturated, and the reactions were incubated at 40°C with shaking at 1000 rpm. Before analysis, soluble products generated by the enzymes were converted to GlcNAc and chitobionic acid by overnight incubation with 1.5 μm SmCHB (a chitobiase) at 37 °C (60). The data points represent the mean of three independent experiments ± S.D. In B, the curves representing JdCBM5-Chi18 and JdChi18 overlap; hence only one (for JdChi18) is visible.