FIG. 3.

Effect of pAPMSF on formation and secretion of some cellulases and xylanases by T. reesei. (A) Preprosequences of the enzymes investigated. The arrows indicate the N-terminal amino acid of the secreted protein according to published evidence (2, 39, 42, 44). (B) Demonstration of the intra- and extracellular levels of these enzymes by SDS-PAGE and immunostaining with specific monoclonal antibodies. Experiments were carried out as described in Materials and Methods. The marks in the left margin indicate the positions of the unprocessed (higher mark) and processed (lower mark) enzyme form. Blots for extracellular and intracellular samples from the same protein are placed so that the relative positions of these two enzyme forms are comparable. For intracellular samples, equal amounts (40 μg) of protein were loaded onto the gels; equal volumes (20 μl) of the culture broth were loaded for the extracellular samples. Only a single mark is shown where only the processed form can be observed (CBH I and CBH II). Results shown are representative of at least three separate experiments.