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

doi:10.1016/B978-0-12-415931-0.00009-4

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 journal › Article › Scientific › peer-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 language	English
Pages (from-to)	169-182
Journal	Methods in Enzymology
Volume	510
DOIs	https://doi.org/10.1016/B978-0-12-415931-0.00009-4
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

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10.1016/B978-0-12-415931-0.00009-4

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@article{dc113cd7f5e8425bb1f90f1c79a2dfef,

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

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.",

keywords = "Biomass utilization, Cellulase, Cellulose, High-speed atomic force microscopy, Single-molecule observations, Trichoderma reesei",

author = "Kiyohiko Igarashi and Takayuki Uchihashi and Anu Koivula and Masahisa Wada and Satoshi Kimura and Merja Penttil{\"a} and Toshio Ando and Masahiro Samejima",

year = "2012",

month = may,

day = "22",

doi = "10.1016/B978-0-12-415931-0.00009-4",

language = "English",

volume = "510",

pages = "169--182",

journal = "Methods in Enzymology",

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T1 - Visualization of cellobiohydrolase I from trichoderma reesei moving on crystalline cellulose using high-speed atomic force microscopy

AU - Igarashi, Kiyohiko

AU - Uchihashi, Takayuki

AU - Koivula, Anu

AU - Wada, Masahisa

AU - Kimura, Satoshi

AU - Penttilä, Merja

AU - Ando, Toshio

AU - Samejima, Masahiro

PY - 2012/5/22

Y1 - 2012/5/22

N2 - 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.

AB - 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.

KW - Biomass utilization

KW - Cellulase

KW - Cellulose

KW - High-speed atomic force microscopy

KW - Single-molecule observations

KW - Trichoderma reesei

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UR - https://www.sciencedirect.com/bookseries/methods-in-enzymology/vol/510/suppl/C

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DO - 10.1016/B978-0-12-415931-0.00009-4

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AN - SCOPUS:84861130110

SN - 0076-6879

VL - 510

SP - 169

EP - 182

JO - Methods in Enzymology

JF - Methods in Enzymology

ER -

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

Abstract

Funding

Keywords

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