The purification and properties of the C1 component of Trichoderma koningii cellulase
- 1 August 1972
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 128 (5), 1183-1192
- https://doi.org/10.1042/bj1281183
Abstract
1. The C1 component that was isolated from a Trichoderma koningii cellulase preparation (Wood, 1968) by chromatography on DEAE-Sephadex with a salt gradient was still associated with a trace of CM-cellulase activity (determined by reducing-sugar and viscometric methods). 2. Further chromatography on DEAE-Sephadex, with a pH gradient instead of a salt gradient, provided a C1 component that could still produce reducing sugars from a solution of CM-cellulose (to a very limited extent), but which could no longer decrease the viscosity (i.e. under the assay conditions employed). 3. No evidence for the non-identity of C1 component and the trace of CM-cellulase activity could be found when electrofocusing was done in a stabilized pH gradient covering three pH units (pH3–6) or, alternatively, only 0.5 pH unit (pH3.72–4.25). 4. The two protein peaks that were separated by electrofocusing in carrier ampholytes covering only 0.5 pH unit (isoelectric pH values of 3.80 and 3.95) were shown to be isoenzymes of the C1 component: they differed in the extent to which they were associated with carbohydrate (9% and 33%). 5. The purified C1 component had little ability to attack CM-cellulose or highly ordered forms of cellulose, but degraded phosphoric acid-swollen cellulose readily: cellobiose was the principal product of the hydrolysis (97%). 6. Dewaxed cotton fibre was degraded to the extent of 15% when exposed to high concentrations of C1 component over a prolonged period: cellobiose was again the principal sugar present in the supernatant (96%). 7. Cellotetraose and cellohexaose were hydrolysed almost exclusively to cellobiose. 8. Evidence indicates that the C1 component is a β-1,4-glucan cellobiosylhydrolase.Keywords
This publication has 16 references indexed in Scilit:
- The cellulase of Fusarium solani. Purification and specificity of the β-(1→4)-glucanase and the β-d-glucosidase componentsBiochemical Journal, 1971
- The formation of short fibres from native cellulose by components of Trichoderma koningii cellulaseBiochemical Journal, 1970
- Another Source of CellulaseNature, 1969
- Extracellular enzyme system utilized by the rot fungus Stereum sanguinolentum for the breakdown of celluloseArchives of Biochemistry and Biophysics, 1968
- The cellulase of Trichoderma viride. Separation of the components involved in the solubilization of cottonBiochemical Journal, 1967
- Isoelectric Fractionation, Analysis, and Characterization of Ampholytes in Natural pH Gradients. IV. Further Studies on the Resolving Power in Connection with Separation of Myoglobins.Acta Chemica Scandinavica, 1966
- PURIFICATION AND CHARACTERIZATION OF TWO TYPES OF CELLULASE FROM TRICHODERMA KONINGI.1964
- EVIDENCE FOR MULTIPLE COMPONENTS IN MICROBIAL CELLULASESCanadian Journal of Microbiology, 1954
- Detection of Sugars on Paper ChromatogramsNature, 1950
- A SUBMICRODETERMINATION OF GLUCOSEJournal of Biological Chemistry, 1949