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

Abstract

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

Authors:
 [1];  [2];  [2];  [3];  [4];  [5];  [6];  [2];  [2];  [2];  [4];  [6]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States)
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  3. Univ. of California, Berkeley, CA (United States)
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
  5. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  6. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23), Biological Systems Science Division (SC-23.2 )
OSTI Identifier:
1511366
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 6; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS

Citation Formats

Reyes-Ortiz, Vimalier, Heins, Richard A., Cheng, Gang, Kim, Edward Y., Vernon, Briana C., Elandt, Ryan B., Adams, Paul D., Sale, Kenneth L., Hadi, Masood Z., Simmons, Blake A., Kent, Michael S., and Tullman-Ercek, Danielle. Addition of a carbohydrate-binding module enhances cellulase penetration into cellulose substrates. United States: N. p., 2013. Web. doi:10.1186/1754-6834-6-93.
Reyes-Ortiz, Vimalier, Heins, Richard A., Cheng, Gang, Kim, Edward Y., Vernon, Briana C., Elandt, Ryan B., Adams, Paul D., Sale, Kenneth L., Hadi, Masood Z., Simmons, Blake A., Kent, Michael S., & Tullman-Ercek, Danielle. Addition of a carbohydrate-binding module enhances cellulase penetration into cellulose substrates. United States. doi:10.1186/1754-6834-6-93.
Reyes-Ortiz, Vimalier, Heins, Richard A., Cheng, Gang, Kim, Edward Y., Vernon, Briana C., Elandt, Ryan B., Adams, Paul D., Sale, Kenneth L., Hadi, Masood Z., Simmons, Blake A., Kent, Michael S., and Tullman-Ercek, Danielle. Wed . "Addition of a carbohydrate-binding module enhances cellulase penetration into cellulose substrates". United States. doi:10.1186/1754-6834-6-93. https://www.osti.gov/servlets/purl/1511366.
@article{osti_1511366,
title = {Addition of a carbohydrate-binding module enhances cellulase penetration into cellulose substrates},
author = {Reyes-Ortiz, Vimalier and Heins, Richard A. and Cheng, Gang and Kim, Edward Y. and Vernon, Briana C. and Elandt, Ryan B. and Adams, Paul D. and Sale, Kenneth L. and Hadi, Masood Z. and Simmons, Blake A. and Kent, Michael S. and Tullman-Ercek, Danielle},
abstractNote = {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. 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. Our findings suggest that the CBM improves the efficiency of these cellulases by enabling digestion within the bulk of the film.},
doi = {10.1186/1754-6834-6-93},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 6,
place = {United States},
year = {2013},
month = {7}
}

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Cited by: 26 works
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Works referenced in this record:

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