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

15 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

Keywords

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

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