Comparative hydrolysis of Avicel by the 14 sets of designer cellulosomes. The modular composition of each set and the scaffoldin number is denoted on the x-axis. Upper panel: the carbohydrate binding module (CBM) of the designer scaffoldin is at one of the terminal positions (N or C terminus). Lower panel: the CBM module of the designer scaffoldin is in an internal position. Each designer-cellulosome set is assembled either without intermodular linker scaffoldin (light brown), with short intermodular linker scaffoldin (medium brown), and with long intermodular linker scaffoldin (dark brown). Controls: Free: corresponds to the combined activity of 48S-t, 9K-a and 8A-b. CBM-Coh represents a cellulose-targeting control, corresponding to the activity of the three dockerin-bearing enzymes, each attached separately to its matching cohesin module fused to a CBM. Reactions were carried out for 72 h. Enzymatic activity was defined by mM reducing sugars as determined by a glucose standard curve. All reactions were carried out in triplicate and repeated three times. Standard deviations of at least three experiments are indicated.