Abstract
Cellulases hydrolyze β-1,4-glucosidic linkages of insoluble cellulose at the solid/liquid interface, generating soluble cellooligosaccharides. We describe here our method for real-time observation of the behavior of cellulase molecules on the substrate, using high-speed atomic force microscopy (HS-AFM). When glycoside hydrolase family 7 cellobiohydrolase from Trichoderma reesei (TrCel7A) was incubated with crystalline cellulose, many enzyme molecules were observed to move unidirectionally on the surface of the substrate by HS-AFM. The velocity of the moving molecules of TrCel7A on cellulose I crystals was estimated by means of image analysis.
Original language | English |
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Pages (from-to) | 169-182 |
Journal | Methods in Enzymology |
Volume | 510 |
DOIs | |
Publication status | Published - 22 May 2012 |
MoE publication type | A1 Journal article-refereed |
Funding
This research was supported by Grants-in-Aid for Scientific Research to K. I. (19688016 and 21688023), T. U. (21023010 and 21681017), and T. A. (20221006) from the Japanese Ministry of Education, Culture, Sports, and Technology; by a grant of the Knowledge Cluster Initiative to T. A.; by a Grant for Development of Technology for High Efficiency Bioenergy Conversion Project to M. S. (07003004-0) from the New Energy and Industrial Technology Development Organization; and by an Advanced Low Carbon Technology Research and Development Program from the Japan Science and Technology Agency to K. I. and T. U.
Keywords
- Biomass utilization
- Cellulase
- Cellulose
- High-speed atomic force microscopy
- Single-molecule observations
- Trichoderma reesei