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

Research output: Contribution to journalArticleScientificpeer-review

11 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

Fingerprint

Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Atomic Force Microscopy
Cellulose
Atomic force microscopy
Visualization
Crystalline materials
Molecules
Cellulases
Cellulase
Glycoside Hydrolases
Substrates
Image analysis
Crystals
Liquids
Enzymes

Keywords

  • Biomass utilization
  • Cellulase
  • Cellulose
  • High-speed atomic force microscopy
  • Single-molecule observations
  • Trichoderma reesei

Cite this

Igarashi, Kiyohiko ; Uchihashi, Takayuki ; Koivula, Anu ; Wada, Masahisa ; Kimura, Satoshi ; Penttilä, Merja ; Ando, Toshio ; Samejima, Masahiro. / Visualization of cellobiohydrolase i from trichoderma reesei moving on crystalline cellulose using high-speed atomic force microscopy. In: Methods in Enzymology. 2012 ; Vol. 510. pp. 169-182.
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Visualization of cellobiohydrolase i from trichoderma reesei moving on crystalline cellulose using high-speed atomic force microscopy. / Igarashi, Kiyohiko; Uchihashi, Takayuki; Koivula, Anu; Wada, Masahisa; Kimura, Satoshi; Penttilä, Merja; Ando, Toshio; Samejima, Masahiro.

In: Methods in Enzymology, Vol. 510, 22.05.2012, p. 169-182.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Igarashi, Kiyohiko

AU - Uchihashi, Takayuki

AU - Koivula, Anu

AU - Wada, Masahisa

AU - Kimura, Satoshi

AU - Penttilä, Merja

AU - Ando, Toshio

AU - Samejima, Masahiro

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KW - Cellulase

KW - Cellulose

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KW - Single-molecule observations

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