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

Vimalier Reyes-Ortiz, Richard A. Heins, Gang Cheng, Edward Y. Kim, Briana C. Vernon, Ryan B. Elandt, Paul D. Adams, Kenneth L. Sale, Masood Z. Hadi, Blake A. Simmons, Michael S. Kent, Danielle Tullman-Ercek*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

Abstract

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.

Original languageEnglish (US)
Article number93
JournalBiotechnology for Biofuels
Volume6
Issue number1
DOIs
StatePublished - 2013

Keywords

  • Carbohydrate-Binding modules
  • Cellulases
  • Cellulose model films
  • Endoglucanases
  • Neutron reflectometry

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • Energy(all)
  • Management, Monitoring, Policy and Law

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