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 | |
| Publication status | Published - 22 May 2012 |
| MoE publication type | A1 Journal article-refereed |
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Keywords
- Biomass utilization
- Cellulase
- Cellulose
- High-speed atomic force microscopy
- Single-molecule observations
- Trichoderma reesei
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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 journal › Article › Scientific › peer-review
TY - JOUR
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
N1 - CA2: TK404 CA2: TK400 SDA: BIC ISI: BIOCHEMISTRY & MOLECULAR BIOLOGY
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
UR - http://www.scopus.com/inward/record.url?scp=84861130110&partnerID=8YFLogxK
UR - https://www.sciencedirect.com/bookseries/methods-in-enzymology/vol/510/suppl/C
U2 - 10.1016/B978-0-12-415931-0.00009-4
DO - 10.1016/B978-0-12-415931-0.00009-4
M3 - Article
C2 - 22608726
AN - SCOPUS:84861130110
VL - 510
SP - 169
EP - 182
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
ER -