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Visualizing cellulase adsorption and quantitatively determining cellulose accessibility with an updated fungal
Tian Li1, Nan Liu1,2, Xianjin Ou3
1Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Institute of Applied Chemistry, Department of Chemical Engineering, Tsinghua University, Beijing, 100084 China.
Biotechnology for Biofuels
|April 17, 2018
Summary
A new fusion protein probe accurately measures cellulose accessibility (CAC) in biomass. This tool helps understand how structural features affect cellulose digestibility and enzyme adsorption.
Area of Science:
- Biotechnology
- Biochemistry
- Materials Science
Background:
- Cellulose accessibility to cellulases (CAC) is crucial for enzymatic digestibility of lignocellulosic biomass.
- Improving CAC via pretreatment is essential for efficient fermentable sugar release.
- Conventional porosimetry methods have limitations for investigating CAC.
Purpose of the Study:
- To develop a novel fusion protein for quantitative determination of CAC.
- To create a versatile tool for assessing biomass structural impacts on cellulose accessibility.
Main Methods:
- Construction of a fusion protein: fungal cellulose-binding module (CBM) from Cel7A and di-green fluorescent protein (GFP2).
- Optimization of fusion protein construction strategies (His-tag site, module order) for expression and usability.
- Preparation of cellulosic substrates with varied hemicellulose, lignin, and formyl group content.
Main Results:
- The His6-CBM-GFP2 fusion protein demonstrated suitability for quantitative CAC analysis.
- CAC values ranged from 0.6–20.4 m²/g, correlating with substrate structural features.
- Identified optimal probe protein for investigating substrate structural effects on cellulose accessibility.
Conclusions:
- The fusion probe protein serves as a versatile tool for quantitative CAC investigation.
- Enables assessment of biomass structural features' impact on cellulose hydrolyzability.
- Facilitates study of nonproductive cellulase enzyme adsorption on lignin.
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