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Characterization of commercial Trichoderma reesei cellulase preparations by denaturing electrophoresis (SDS‐PAGE) and immunostaining using monoclonal antibodies
Author(s) -
KubicekPranz EM,
Gsur A.,
Hayn M.,
Kubicek CP
Publication year - 1991
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1991.tb00184.x
Subject(s) - trichoderma reesei , cellulase , gel electrophoresis , polyacrylamide gel electrophoresis , chemistry , biochemistry , monoclonal antibody , chromatography , sodium dodecyl sulfate , protease , cellulose , beta glucosidase , microbiology and biotechnology , enzyme , biology , antibody , immunology
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.