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

Paper ID Volume ID Publish Year Pages File Format Full-Text
17218 42652 2012 6 PDF Available
Title
A simple and fast method for the determination of endo- and exo-cellulase activity in cellulase preparations using filter paper
Abstract

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 (FPAinsol) 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.

► Selective determination of endo- (EG) and exo- (ExG) cellulase activity on filter paper. ► Enzymatic cellulose hydrolysis starts with insoluble reducing sugar formation. ► The new procedure allows to calculate the endo/exo activity ratio of cellulase preparations. ► EG activity measured on filter paper provides different information than CMC activity.

Keywords
Cellulases; Endoglucanase activity; Exoglucanase activity; Filter paper activity; Endo–exo activity ratio; Hypocrea jecorina (Trichoderma reesei)
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A simple and fast method for the determination of endo- and exo-cellulase activity in cellulase preparations using filter paper
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 51, Issue 5, 10 October 2012, Pages 280–285
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us