FIG 5.

Effect of sequential additions of TfCel48A and TfCel9A on each other's activities. (a) BC was pretreated with either TfCel48A or TfCel9A for 20 h, and the reaction mixtures were boiled to denature the enzyme. The synergistic partner was then added, and the reaction mixtures were incubated for 2.25 h. Comparisons of activity on untreated and enzyme-pretreated substrate are provided. For the pretreated reaction mixtures, the provided value was adjusted by subtracting the amount of soluble sugar present in the reaction mixture measured after the pretreatment from the total amount of sugar measured at the end of the assay. (b) Initial TfCel9A (83 nM) digestion was followed by the addition of TfCel9A (83 nM) with or without an intermediate incubation with TfCel48A (750 nM). The intermediate addition of TfCel48A enhanced the activity of the freshly added TfCel9A, as evidenced by the steeper percent digestion profile following incubation with TfCel48A.