A simple and fast method for the determination of endo- and exo-cellulase activity in cellulase preparations using filter paper.

Marcos Henrique Luciano Silveira, Martinho Rau, Elba Pinto da Silva Bon, Jürgen Andreaus

Department of Chemistry, Regional University of Blumenau, Rua Antônio da Veiga, 140, 89010-971 Blumenau, SC, Brazil.

Enzyme and microbial technology 2012 Oct 10

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This study describes a procedure for the selective determination of endo- (EG) and exo- (ExG) cellulase activities using filter paper as the sole substrate. The procedure is based on the enzymes mode of action whereby EG activity predominantly forms insoluble reducing sugars and ExG activity soluble reducing sugars. The procedure was developed using filter paper as substrate for hydrolysis with three cellulase preparations of Hypocrea jecorina containing either endoglucanase (EG), predominantly exoglucanase (ExG) or both endo- and exoglucanase activities. Hydrolysis experiments, which were followed assessing the formation of total, soluble and insoluble reducing sugars (RS), showed that up to 30 min of hydrolysis predominantly insoluble reducing sugars were formed, while after this initial hydrolysis stage soluble reducing sugar formation increased significantly, making it thus possible to measure separately EG and ExG activity. FPA activities obtained from the reaction products at different reaction times suggest that EG-activity (FPA(insol)) should be measured between 10 and 20 min of hydrolysis. The proposed procedure allows to evaluate the EG and ExG activity contribution to total cellulase activity and to calculate the endo/exo activity ratio of any cellulase preparation. Copyright © 2012 Elsevier Inc. All rights reserved.

Citation

Marcos Henrique Luciano Silveira, Martinho Rau, Elba Pinto da Silva Bon, Jürgen Andreaus. A simple and fast method for the determination of endo- and exo-cellulase activity in cellulase preparations using filter paper. Enzyme and microbial technology. 2012 Oct 10;51(5):280-5