Quantification of cellulase activity using cellulose-azure

Despite wide application of cellulose-azure as a substrate for measuring cellulase activity, there is no quantification of hydrolysis rate or enzymatic activities using this substrate. The aim of this study was to quantify the hydrolysis rate in terms of product formation and dye released using cellulose-azure. The amount of dye released was correlated with the production of glucose and the enzyme concentrations. It is shown that the lack of correlation can be due to (1) repression of the release of the azure-dye when azure-dye accumulates, (2) presence of degradable substrates in the cellulase powder which inflate the glucose measurements and (3) the degradation of cellulose which is not linked to the dye in the cellulose-azure. Based on the lack of correlation, it is recommended that cellulose-azure should only be applied in assays when the aim is to compare relative activities of different enzymatic systems.