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Title: Selective fluorescence labeling: time-lapse enzyme visualization during sugarcane hydrolysis
Authors: Imai, Makiko
Mihashi, Asako
Imai, Tomoya
Kimura, Satoshi
Matsuzawa, Tomohiko
Yaoi, Katsuro
Shibata, Nozomu
Kakeshita, Hiroshi
Igarashi, Kazuaki
Kobayashi, Yoshinori
Sugiyama, Junji  kyouindb  KAKEN_id
Author's alias: 今井, 牧子
今井, 友也
木村, 聡
杉山, 淳司
Keywords: Cellulose
Sugarcane tissue
Fluorescence microscopy
Statistical image analysis
Issue Date: 27-Mar-2019
Publisher: Springer Nature
Journal title: Journal of Wood Science
Volume: 65
Thesis number: 17
Abstract: Enzymatic biomass saccharification is an important process for bioethanol production. Hitherto, numerous cellulase cocktails (crude enzyme) have been developed to improve enzymatic activity. For this purpose, the synergy of incorporating hydrolase functionality within a cellulase cocktail is a key function. However, such synergistic action, by potentially numerous different enzyme types, on biomass tissue has not been considered despite the importance toward the realistic case of biomass saccharification. This study aims to visualize the behavior of each of the key cellulase components on biomass tissue during saccharification. Time-lapse fluorescence microscopy observations were conducted during saccharification of a thin transverse sugarcane section to monitor enzymes modified with a fluorescence dye. Statistical image analysis successfully demonstrated a unique adsorption/desorption behavior of each enzyme component. Particularly, the behavior of endoxylanase10 (Xyn10), which was recently discovered from Penicillium sp. as a high-performance xylanase, displayed remarkable adsorption on tissues of sugarcane, which accounts for the superior activity of the cellulase mixture with Xyn10.
Rights: © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
DOI(Published Version): 10.1186/s10086-019-1798-0
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