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

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

doi:10.1186/1754-6834-6-93

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

Journal Article · Wed Jul 03 04:00:00 EDT 2013 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/1754-6834-6-93· OSTI ID:1511366

Reyes-Ortiz, Vimalier ^[1]; Heins, Richard A. ^[2]; Cheng, Gang ^[2]; Kim, Edward Y. ^[3]; Vernon, Briana C. ^[4]; Elandt, Ryan B. ^[5]; Adams, Paul D. ^[6]; Sale, Kenneth L. ^[2]; Hadi, Masood Z. ^[2]; Simmons, Blake A. ^[2]; Kent, Michael S. ^[4]; Tullman-Ercek, Danielle ^[6]

Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
Univ. of California, Berkeley, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23), Biological Systems Science Division (SC-23.2 )

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1511366

Alternate ID(s):: OSTI ID: 1153429

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 6; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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