skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Engineering enhanced cellobiohydrolase activity

Abstract

Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure–activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [3];  [1]; ORCiD logo [1];  [1]; ORCiD logo [1]; ORCiD logo [2]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
  3. National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center; Inst. of Advanced Study in Science and Technology (IASST), Guwahati (India). Life Sciences Division
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1432453
Report Number(s):
NREL/JA-2700-70605
Journal ID: ISSN 2041-1723
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; hydrolases; polysaccharides; protein design; x-ray crystallography

Citation Formats

Taylor, Larry E., Knott, Brandon C., Baker, John O., Alahuhta, P. Markus, Hobdey, Sarah E., Linger, Jeffrey G., Lunin, Vladimir V., Amore, Antonella, Subramanian, Venkataramanan, Podkaminer, Kara, Xu, Qi, VanderWall, Todd A., Schuster, Logan A., Chaudhari, Yogesh B., Adney, William S., Crowley, Michael F., Himmel, Michael E., Decker, Stephen R., and Beckham, Gregg T. Engineering enhanced cellobiohydrolase activity. United States: N. p., 2018. Web. doi:10.1038/s41467-018-03501-8.
Taylor, Larry E., Knott, Brandon C., Baker, John O., Alahuhta, P. Markus, Hobdey, Sarah E., Linger, Jeffrey G., Lunin, Vladimir V., Amore, Antonella, Subramanian, Venkataramanan, Podkaminer, Kara, Xu, Qi, VanderWall, Todd A., Schuster, Logan A., Chaudhari, Yogesh B., Adney, William S., Crowley, Michael F., Himmel, Michael E., Decker, Stephen R., & Beckham, Gregg T. Engineering enhanced cellobiohydrolase activity. United States. doi:10.1038/s41467-018-03501-8.
Taylor, Larry E., Knott, Brandon C., Baker, John O., Alahuhta, P. Markus, Hobdey, Sarah E., Linger, Jeffrey G., Lunin, Vladimir V., Amore, Antonella, Subramanian, Venkataramanan, Podkaminer, Kara, Xu, Qi, VanderWall, Todd A., Schuster, Logan A., Chaudhari, Yogesh B., Adney, William S., Crowley, Michael F., Himmel, Michael E., Decker, Stephen R., and Beckham, Gregg T. Thu . "Engineering enhanced cellobiohydrolase activity". United States. doi:10.1038/s41467-018-03501-8. https://www.osti.gov/servlets/purl/1432453.
@article{osti_1432453,
title = {Engineering enhanced cellobiohydrolase activity},
author = {Taylor, Larry E. and Knott, Brandon C. and Baker, John O. and Alahuhta, P. Markus and Hobdey, Sarah E. and Linger, Jeffrey G. and Lunin, Vladimir V. and Amore, Antonella and Subramanian, Venkataramanan and Podkaminer, Kara and Xu, Qi and VanderWall, Todd A. and Schuster, Logan A. and Chaudhari, Yogesh B. and Adney, William S. and Crowley, Michael F. and Himmel, Michael E. and Decker, Stephen R. and Beckham, Gregg T.},
abstractNote = {Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure–activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.},
doi = {10.1038/s41467-018-03501-8},
journal = {Nature Communications},
issn = {2041-1723},
number = ,
volume = 9,
place = {United States},
year = {2018},
month = {3}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 4 works
Citation information provided by
Web of Science

Save / Share: