Figure 3

Dose and time responses of CPH substrates. Reaction plate containing four different coloured versions of CPH-galactomannan (GALMAN) - red, blue, yellow and green. (B) Product plate containing the products of the digestion of the substrates shown in (A) with mannanase (man) used in the range of 0 to 2 U/mL (shown to the right of (A) in 100 mM potassium phosphate buffer pH 7.0 for 1 h at room temperature. (C) Graph showing the absorbance of products released by treatment with mannanase (man) at the concentrations shown on the x-axis (same conditions as in (B) at wavelengths (shown in the legend) appropriate for each product. (D) Graph showing the absorbance (y-axis to the left) of products released over a time period (from 0 to 100 min) by treatment of four differently coloured versions of CPH-xyloglucan with xyloglucanase (xg) at 0.25 U/mL in 100 mM sodium acetate buffer, pH 5.5. Also shown on this graph (y-axis to the right) is the production of reducing ends over the same time period from undyed xyloglucan, as measured using the 3-methyl-2-benzothiazolinone hydrazone (MBTH) method. Note that an undyed xyloglucan hydrogel was used since the dye would likely interfere with the MBTH method. See Tables 1 and 2 for details of the substrates and enzyme used.