Visualization of cellobiohydrolase I from trichoderma reesei moving on crystalline cellulose using high-speed atomic force microscopy

Kiyohiko Igarashi*, Takayuki Uchihashi, Anu Koivula, Masahisa Wada, Satoshi Kimura, Merja Penttilä, Toshio Ando, Masahiro Samejima

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    25 Citations (Scopus)

    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 languageEnglish
    Pages (from-to)169-182
    JournalMethods in Enzymology
    Volume510
    DOIs
    Publication statusPublished - 22 May 2012
    MoE publication typeA1 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

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