Addition of a carbohydrate-binding module enhances cellulase penetration into cellulose substrates

Biotechnology for Biofuels

Vimalier Reyes-Ortiz⁶  Richard A Heins²  Gang Cheng²  Edward Y Kim⁵  Briana C Vernon¹  Ryan B Elandt³  Paul D Adams⁴  Kenneth L Sale²  Masood Z Hadi²  Blake A Simmons²  Michael S Kent¹  Danielle Tullman-Ercek⁴ 

[1] Sandia National Laboratories, Albuquerque, NM 87185, US

[2] Sandia National Laboratories, Livermore, CA 94550, US

[3] Deconstruction Division, Joint BioEnergy Institute, Emeryville, CA 94608, US

[4] Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, US

[5] Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, US

[6] Department of Bioengineering, University of California, Berkeley, CA 94720, US

关键词: Neutron reflectometry;    Cellulose model films;    Carbohydrate-Binding modules;    Endoglucanases;    Cellulases;   

Others  :  798003
DOI  :  10.1186/1754-6834-6-93

 received in 2012-12-10, accepted in 2013-06-18,  发布年份 2013

【 摘 要 】

Introduction

Cellulases are of great interest for application in biomass degradation, yet the molecular details of the mode of action of glycoside hydrolases during degradation of insoluble cellulose remain elusive. To further improve these enzymes for application at industrial conditions, it is critical to gain a better understanding of not only the details of the degradation process, but also the function of accessory modules.

Method

We fused a carbohydrate-binding module (CBM) from family 2a to two thermophilic endoglucanases. We then applied neutron reflectometry to determine the mechanism of the resulting enhancements.

Results

Catalytic activity of the chimeric enzymes was enhanced up to three fold on insoluble cellulose substrates as compared to wild type. Importantly, we demonstrate that the wild type enzymes affect primarily the surface properties of an amorphous cellulose film, while the chimeras containing a CBM alter the bulk properties of the amorphous film.

Conclusion

Our findings suggest that the CBM improves the efficiency of these cellulases by enabling digestion within the bulk of the film.

【 授权许可】

   
2013 Reyes-Ortiz et al.; licensee BioMed Central Ltd.

【 图 表 】

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