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Detection and differentiation of cellulase components using low molecular mass fluorogenic substrates
Author(s) -
van Tilbeurgh Herman,
Claeyssens Marc
Publication year - 1985
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(85)81260-6
Subject(s) - cellobiose , cellulase , trichoderma reesei , chemistry , chromatography , isoelectric focusing , elution , substrate (aquarium) , lactose , glucosidases , biochemistry , monosaccharide , cellulose , glycoside hydrolase , enzyme , biology , ecology
The 4‐methylumbelliferyl β‐D‐glycosides of glucose, cellobiose, cellotriose and lactose are used to differentiate several exo‐cellobiohydrolase, endocellulase and β‐glucosidase activities in crude cellulase from Trichoderma reesei . Spectrophotometric or fluorimetric assays allow simple detection and quantitative measurements of eluate activities from ion‐exchange chromatography and after analytical gel electrofocusing. Using the fluorophoric glucoside several β‐glucosidases can be visualised after isoelectric focusing on polyacrylamide gels. The use of the lactoside and of the same substrate supplemented with cellobiose as inhibitor allows a clearcut distinction to be made between endocellulase II and exo‐cellobiohydrolase I. Both enzymes are present as ‘iso‐enzyme’ mixtures. With the cellotrioside only one fraction is detectable (endocellulase III). The same methods could be used in culture growth experiments.