Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome
Abstract
The ability to deconstruct plant biomass without conventional pretreatment has made members of the genus Caldicellulosiruptor the target of investigation for the consolidated processing of plant lignocellulosic biomass to biofuels and bioproducts. To investigate the synergy of enzymes involved and to further improve the ability of C. bescii to degrade cellulose, we introduced CAZymes that act synergistically with the C. besciii exoproteome in vivo and in vitro. We recently demonstrated that the Acidothermus cellulolyticus E1 endo-1,4-β-D-glucanase (GH5) with a family 2 carbohydrate-binding module (CBM) increased the activity of C. bescii exoproteome on biomass, presumably acting in concert with CelA. The β-glucanase, GuxA, from A. cellulolyticus is a multi-domain enzyme with strong processive exoglucanase activity, and the cellobiose phosphorylase from Thermotoga maritima catalyzes cellulose degradation acting synergistically with cellobiohydrolases and endoglucanases. We identified new chromosomal insertion sites to co-express these enzymes and the resulting strain showed a significant increase in the enzymatic activity of the exoproteome.
- Authors:
-
- 0000 0004 1936 738X grid.213876.9 Department of Genetics University of Georgia 30602 Athens GA USA, 0000 0001 0789 9563 grid.254224.7 Department of Food Science and Technology Chung-Ang University 17546 Anseong Gyeonggi Republic of Korea, 0000 0004 0446 2659 grid.135519.a The BioEnergy Science Center and the Center for BioEnergy Innovation Oak Ridge National Laboratory 37831 Oak Ridge TN USA
- 0000 0001 2199 3636 grid.419357.d Biosciences Center, National Renewable Energy Laboratory Golden CO USA, 0000 0004 0446 2659 grid.135519.a The BioEnergy Science Center and the Center for BioEnergy Innovation Oak Ridge National Laboratory 37831 Oak Ridge TN USA
- 0000 0004 1936 738X grid.213876.9 Department of Genetics University of Georgia 30602 Athens GA USA, 0000 0004 0446 2659 grid.135519.a The BioEnergy Science Center and the Center for BioEnergy Innovation Oak Ridge National Laboratory 37831 Oak Ridge TN USA
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1772499
- Alternate Identifier(s):
- OSTI ID: 1505928
- Report Number(s):
- NREL/JA-2700-73412
Journal ID: ISSN 1367-5435
- Grant/Contract Number:
- AC05-00OR22725; AC36-08GO28308
- Resource Type:
- Published Article
- Journal Name:
- Journal of Industrial Microbiology and Biotechnology
- Additional Journal Information:
- Journal Name: Journal of Industrial Microbiology and Biotechnology Journal Volume: 46 Journal Issue: 5; Journal ID: ISSN 1367-5435
- Publisher:
- Oxford University Press
- Country of Publication:
- Germany
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; consolidated bioprocessing; biomass deconstruction; glucanase; cellobiose phosphorylase; gene integration; Caldicellulosiruptor
Citation Formats
Kim, Sun-Ki, Chung, Daehwan, Himmel, Michael E., Bomble, Yannick J., and Westpheling, Janet. Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome. Germany: N. p., 2019.
Web. doi:10.1007/s10295-019-02150-0.
Kim, Sun-Ki, Chung, Daehwan, Himmel, Michael E., Bomble, Yannick J., & Westpheling, Janet. Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome. Germany. https://doi.org/10.1007/s10295-019-02150-0
Kim, Sun-Ki, Chung, Daehwan, Himmel, Michael E., Bomble, Yannick J., and Westpheling, Janet. Wed .
"Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome". Germany. https://doi.org/10.1007/s10295-019-02150-0.
@article{osti_1772499,
title = {Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome},
author = {Kim, Sun-Ki and Chung, Daehwan and Himmel, Michael E. and Bomble, Yannick J. and Westpheling, Janet},
abstractNote = {The ability to deconstruct plant biomass without conventional pretreatment has made members of the genus Caldicellulosiruptor the target of investigation for the consolidated processing of plant lignocellulosic biomass to biofuels and bioproducts. To investigate the synergy of enzymes involved and to further improve the ability of C. bescii to degrade cellulose, we introduced CAZymes that act synergistically with the C. besciii exoproteome in vivo and in vitro. We recently demonstrated that the Acidothermus cellulolyticus E1 endo-1,4-β-D-glucanase (GH5) with a family 2 carbohydrate-binding module (CBM) increased the activity of C. bescii exoproteome on biomass, presumably acting in concert with CelA. The β-glucanase, GuxA, from A. cellulolyticus is a multi-domain enzyme with strong processive exoglucanase activity, and the cellobiose phosphorylase from Thermotoga maritima catalyzes cellulose degradation acting synergistically with cellobiohydrolases and endoglucanases. We identified new chromosomal insertion sites to co-express these enzymes and the resulting strain showed a significant increase in the enzymatic activity of the exoproteome.},
doi = {10.1007/s10295-019-02150-0},
journal = {Journal of Industrial Microbiology and Biotechnology},
number = 5,
volume = 46,
place = {Germany},
year = {Wed May 01 00:00:00 EDT 2019},
month = {Wed May 01 00:00:00 EDT 2019}
}
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Expression of a Cellobiose Phosphorylase from Thermotoga maritima in Caldicellulosiruptor bescii Improves the Phosphorolytic Pathway and Results in a Dramatic Increase in Cellulolytic Activity
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Works referencing / citing this record:
Genomic and physiological analyses reveal that extremely thermophilic Caldicellulosiruptor changbaiensis deploys uncommon cellulose attachment mechanisms
journal, August 2019
- Khan, Asma M. A. M.; Mendoza, Carl; Hauk, Valerie J.
- Journal of Industrial Microbiology & Biotechnology, Vol. 46, Issue 9-10