The Mechanism of Enzymatic Cellulose Degradation
Purification of a Cellulolytic Enzyme from Trichoderma viride Active on Highly Ordered Cellulose
Article first published online: 3 MAR 2005
European Journal of Biochemistry
Volume 37, Issue 1, pages 21–30, August 1973
How to Cite
Berghem, L. E. R. and Pettersson, L. G. (1973), The Mechanism of Enzymatic Cellulose Degradation. European Journal of Biochemistry, 37: 21–30. doi: 10.1111/j.1432-1033.1973.tb02952.x
- Issue published online: 3 MAR 2005
- Article first published online: 3 MAR 2005
- (Received December 7, 1972/May 2, 1973)
- 1A cellulolytic enzyme (“C1” enzyme) has been isolated from a commercial cellulase preparation derived from culture filtrates of the fungus Trichoderma viride.
- 2The purification method is a four-step procedure including chromatography on Bio-Gel P-10, DEAE-Sephadex chromatography, isoelectric focusing and chromatography on Bio-Gel P-60.
- 3A yield of 144 mg enzyme was obtained per 100 g commercial cellulase.
- 4The isolated enzyme was homogeneous in polyacrylamide gel electrophoresis at pH 5.0 and at pH 8.0 by isoelectric focusing in a polyacrylamide gel and also in the ultracentrifuge.
- 5No enzyme activity towards carboxymethylcellulose could be detected in the purified material under the assay conditions used. Similarly, there was no β-glucosidase activity.
- 6The purified enzyme was associated with 3.3% carbohydrate and is assumed to be a glycoprotein. The enzyme was isoelectric at pH 3.79 (10°C). A molecular weight of 46000 was determined by chromatography of the reduced and alkylated enzyme on a calibrated column of Sepharose 6B in 6 M guanidine-HCl.
- 7Crystalline cellulose (Avicel), phosphoric acid-swollen Avicel and cellotetraose were degraded by the enzyme and in each case the principle reaction product was cellobiose.
- 8Evidence indicates that the purified enzyme is a β-1,4-glucan cellobiohydrolase.