title: Characterization of commercial Trichoderma reesei cellulase preparations by denaturing electrophoresis (SDS-PAGE) and immunostaining using monoclonal antibodies.
abstract: Fifteen different cellulase preparations from Trichoderma reesei, obtained either commercially or from pilot plants, were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting using monoclonal antibodies against two cellobiohydrolases (CBH I, CBH II), an endoglucanase (EG I), and beta-glucosidase. The staining patterns were compared with the activities of the preparations against filter paper (FPU), carboxymethylcellulose (CMC-ase), cellobiose (beta-glucosidase), and azocasein (protease). Variable amounts of proteolytic degradation products of CBH I, CBH II, and EG I were seen in most samples, and only half of them contained intact beta-glucosidase. The degree of proteolysis did not correlate with any significant difference in the respective activities of these preparations against filter paper cellulose or carboxymethylcellulose. In more than 50% of all cases a decreased beta-glucosidase activity and the absence of intact beta-glucosidase protein in Western blots was observed in preparations displaying high proteolytic activity.
authors.affiliation: Abteilung für Mikrobielle Biochemie, TU Wien, Austria.
authors.forename: E M
authors.initials: EM
authors.lastname: Kubicek-Pranz
authors.forename: A
authors.initials: A
authors.lastname: Gsur
authors.forename: M
authors.initials: M
authors.lastname: Hayn
authors.forename: C P
authors.initials: CP
authors.lastname: Kubicek
mesh_terms: D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002480:Cellulase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D011489:Protein Denaturation; D014242:Trichoderma / Q000201:enzymology*
chemical_list: D000911:Antibodies, Monoclonal; D002480:Cellulase