Characterization of commercial Trichoderma reesei cellulase preparations by denaturing electrophoresis (SDS-PAGE) and immunostaining using monoclonal antibodies.
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.
Kubicek-Pranz
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EM
Abteilung für Mikrobielle Biochemie, TU Wien, Austria.
Gsur
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Hayn
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Kubicek
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CP
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*
D000911:Antibodies, Monoclonal; D002480:Cellulase