Share Email Print
cover

Proceedings Paper

A single molecule study of cellulase hydrolysis of crystalline cellulose
Author(s): Yu-San Liu; Yonghua Luo; John O. Baker; Yining Zeng; Michael E. Himmel; Steve Smith; Shi-You Ding
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper Abstract

Cellobiohydrolase-I (CBH I), a processive exoglucanase secreted by Trichoderma reesei, is one of the key enzyme components in a commercial cellulase mixture currently used for processing biomass to biofuels. CBH I contains a family 7 glycoside hydrolase catalytic module, a family 1 carbohydrate-binding module (CBM), and a highlyglycosylated linker peptide. It has been proposed that the CBH I cellulase initiates the hydrolysis from the reducing end of one cellulose chain and successively cleaves alternate β-1,4-glycosidic bonds to release cellobiose as its principal end product. The role each module of CBH I plays in the processive hydrolysis of crystalline cellulose has yet to be convincingly elucidated. In this report, we use a single-molecule approach that combines optical (Total Internal Reflection Fluorescence microscopy, or TIRF-M) and non-optical (Atomic Force Microscopy, or AFM) imaging techniques to analyze the molecular motion of CBM tagged with green fluorescence protein (GFP), and to investigate the surface structure of crystalline cellulose and changes made in the structure by CBM and CBH I. The preliminary results have revealed a confined nanometer-scale movement of the TrCBM1-GFP bound to cellulose, and decreases in cellulose crystal size as well as increases in surface roughness during CBH I hydrolysis of crystalline cellulose.

Paper Details

Date Published: 25 February 2010
PDF: 8 pages
Proc. SPIE 7571, Single Molecule Spectroscopy and Imaging III, 757103 (25 February 2010); doi: 10.1117/12.840975
Show Author Affiliations
Yu-San Liu, National Renewable Energy Lab. (United States)
Oak Ridge National Lab. (United States)
Yonghua Luo, National Renewable Energy Lab. (United States)
John O. Baker, National Renewable Energy Lab. (United States)
Yining Zeng, National Renewable Energy Lab. (United States)
Oak Ridge National Lab. (United States)
Michael E. Himmel, National Renewable Energy Lab. (United States)
Oak Ridge National Lab. (United States)
Steve Smith, South Dakota School of Mines and Technology (United States)
Shi-You Ding, National Renewable Energy Lab. (United States)
Oak Ridge National Lab. (United States)


Published in SPIE Proceedings Vol. 7571:
Single Molecule Spectroscopy and Imaging III
Jörg Enderlein; Zygmunt Karol Gryczynski; Rainer Erdmann, Editor(s)

© SPIE. Terms of Use
Back to Top