Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass
Abstract
Background: Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic organisms described to date, and have the ability to grow on lignocellulosic biomass without conventional pretreatment. Different species vary in their abilities to degrade cellulose, and the presence of CelA, a bifunctional glycoside hydrolase that contains a Family 48 and a Family 9 catalytic domain, correlates well with cellulolytic ability in members of this genus. For example, C. hydrothermalis, which does not contain a CelA homolog, or a GH48 Family or GH9 Family glycoside hydrolase, is the least cellulolytic of the Caldicellulosiruptor species so far described. C. bescii, which contains CelA and expresses it constitutively, is among the most cellulolytic. In fact, CelA is the most abundant extracellular protein produced in C. bescii. The enzyme contains two catalytic units, a Family 9A-CBM3c processive endoglucanase and a Family 48 exoglucanase, joined by two Family 3b carbohydrate-binding domains. Although there are two non-reducing end-specific Family 9 and three reducing end-specific Family 48 glycoside hydrolases (producing primarily glucose and cellobiose; and cellobiose and cellotriose, respectively) in C. bescii, CelA is the only protein that combines both enzymatic activities. Results: A deletion of the celA gene resulted in a dramatic reduction inmore »
- Authors:
-
- Univ. of Georgia, Athens, GA (United States). Department of Genetics; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center
- National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center
- National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center
- Publication Date:
- Research Org.:
- National Renewable Energy Lab. (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1163406
- Report Number(s):
- NREL/JA-2700-63134
Journal ID: ISSN 1754-6834; MainId:18098;UUID:7f13f9ac-c95b-e411-b769-d89d67132a6d;MainAdminID:6576
- DOE Contract Number:
- AC36-08GO28308
- Resource Type:
- Program Document
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; bioenergy; cellulase; thermophile
Citation Formats
Young, Jenna, Chung, Daehwan, Bomble, Yannick J., Himmel, Michael E., and Westpheling, Janet. Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass. United States: N. p., 2014.
Web. doi:10.1186/s13068-014-0142-6.
Young, Jenna, Chung, Daehwan, Bomble, Yannick J., Himmel, Michael E., & Westpheling, Janet. Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass. United States. https://doi.org/10.1186/s13068-014-0142-6
Young, Jenna, Chung, Daehwan, Bomble, Yannick J., Himmel, Michael E., and Westpheling, Janet. 2014.
"Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass". United States. https://doi.org/10.1186/s13068-014-0142-6. https://www.osti.gov/servlets/purl/1163406.
@article{osti_1163406,
title = {Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass},
author = {Young, Jenna and Chung, Daehwan and Bomble, Yannick J. and Himmel, Michael E. and Westpheling, Janet},
abstractNote = {Background: Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic organisms described to date, and have the ability to grow on lignocellulosic biomass without conventional pretreatment. Different species vary in their abilities to degrade cellulose, and the presence of CelA, a bifunctional glycoside hydrolase that contains a Family 48 and a Family 9 catalytic domain, correlates well with cellulolytic ability in members of this genus. For example, C. hydrothermalis, which does not contain a CelA homolog, or a GH48 Family or GH9 Family glycoside hydrolase, is the least cellulolytic of the Caldicellulosiruptor species so far described. C. bescii, which contains CelA and expresses it constitutively, is among the most cellulolytic. In fact, CelA is the most abundant extracellular protein produced in C. bescii. The enzyme contains two catalytic units, a Family 9A-CBM3c processive endoglucanase and a Family 48 exoglucanase, joined by two Family 3b carbohydrate-binding domains. Although there are two non-reducing end-specific Family 9 and three reducing end-specific Family 48 glycoside hydrolases (producing primarily glucose and cellobiose; and cellobiose and cellotriose, respectively) in C. bescii, CelA is the only protein that combines both enzymatic activities. Results: A deletion of the celA gene resulted in a dramatic reduction in the microorganism’s ability to grow on crystalline cellulose (Avicel) and diminished growth on lignocellulosic biomass. A comparison of the overall endoglucanase and exoglucanase activities of the mutant compared with the wild-type suggests that the loss of the endoglucanase activity provided by the GH9 family domain is perhaps compensated for by other enzymes produced by the cell. In contrast, it appears that no other enzymes in the C. bescii secretome can compensate for the loss of exoglucanase activity. The change in enzymatic activity in the celA mutant resulted in a 15-fold decrease in sugar release on Avicel compared with the parent and wild-type strains. In conclusion: The exoglucanase activity of the GH48 domain of CelA plays a major role in biomass degradation within the suite of C. bescii biomass-degrading enzymes.},
doi = {10.1186/s13068-014-0142-6},
url = {https://www.osti.gov/biblio/1163406},
journal = {Biotechnology for Biofuels},
issn = {1754-6834},
number = 1,
volume = 7,
place = {United States},
year = {Thu Oct 09 00:00:00 EDT 2014},
month = {Thu Oct 09 00:00:00 EDT 2014}
}
Works referenced in this record:
Identification and characterization of CbeI, a novel thermostable restriction enzyme from Caldicellulosiruptor bescii DSM 6725 and a member of a new subfamily of HaeIII-like enzymes
journal, May 2011
- Chung, Dae-Hwan; Huddleston, Jennifer R.; Farkas, Joel
- Journal of Industrial Microbiology & Biotechnology, Vol. 38, Issue 11
Deletion of the Cel48S cellulase from Clostridium thermocellum
journal, September 2010
- Olson, Daniel G.; Tripathi, Shital A.; Giannone, Richard J.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 41, p. 17727-17732
Degradation of microcrystalline cellulose and non-pretreated plant biomass by a cell-free extracellular cellulase/hemicellulase system from the extreme thermophilic bacterium Caldicellulosiruptor bescii
journal, January 2013
- Kanafusa-Shinkai, Sumiyo; Wakayama, Jun'ichi; Tsukamoto, Kazumi
- Journal of Bioscience and Bioengineering, Vol. 115, Issue 1
Designing the deconstruction of plant cell walls
journal, June 2008
- Mccann, M.; Carpita, N.
- Current Opinion in Plant Biology, Vol. 11, Issue 3
Isolation and bioinformatic analysis of a novel transposable element, ISCbe4, from the hyperthermophilic bacterium, Caldicellulosiruptor bescii
journal, October 2013
- Cha, Minseok; Wang, Hao; Chung, Daehwan
- Journal of Industrial Microbiology & Biotechnology, Vol. 40, Issue 12
Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii
journal, June 2014
- Chung, D.; Cha, M.; Guss, A. M.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 24, p. 8931-8936
Targeted gene inactivation in Clostridium phytofermentans shows that cellulose degradation requires the family 9 hydrolase Cphy3367: Clostridium phytofermentans hydrolase genetics
journal, December 2009
- Tolonen, Andrew C.; Chilaka, Amanda C.; Church, George M.
- Molecular Microbiology, Vol. 74, Issue 6
Caldicellulosiruptor Core and Pangenomes Reveal Determinants for Noncellulosomal Thermophilic Deconstruction of Plant Biomass
journal, May 2012
- Blumer-Schuette, S. E.; Giannone, R. J.; Zurawski, J. V.
- Journal of Bacteriology, Vol. 194, Issue 15
Insights into plant biomass conversion from the genome of the anaerobic thermophilic bacterium Caldicellulosiruptor bescii DSM 6725
journal, January 2011
- Dam, Phuongan; Kataeva, Irina; Yang, Sung-Jae
- Nucleic Acids Research, Vol. 39, Issue 8
Extremely thermophilic microorganisms for biomass conversion status and prospects
journal, June 2008
- Blumer-Schuette, Sara E.; Kataeva, Irina; Westpheling, Janet
- Current Opinion in Biotechnology, Vol. 19, Issue 3, p. 210-217
Revealing Nature's Cellulase Diversity The Digestion Mechanism of Caldicellulosiruptor bescii CelA
journal, December 2013
- Brunecky, R.; Alahuhta, M.; Xu, Q.
- Science, Vol. 342, Issue 6165, p. 1513-1516
New insights into enzymatic hydrolysis of heterogeneous cellulose by using carbohydrate-binding module 3 containing GFP and carbohydrate-binding module 17 containing CFP
journal, January 2014
- Gao, Shuhong; You, Chun; Renneckar, Scott
- Biotechnology for Biofuels, Vol. 7, Issue 1
Consolidated bioprocessing of cellulosic biomass: an update
journal, October 2005
- Lynd, Lee R.; van Zyl, Willem H.; McBride, John E.
- Current Opinion in Biotechnology, Vol. 16, Issue 5, p. 577-583
Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production
journal, February 2007
- Himmel, M. E.; Ding, S.-Y.; Johnson, D. K.
- Science, Vol. 315, Issue 5813, p. 804-807
Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance
journal, January 2010
- Park, Sunkyu; Baker, John O.; Himmel, Michael E.
- Biotechnology for Biofuels, Vol. 3, Issue 1
Carbohydrate and lignin are simultaneously solubilized from unpretreated switchgrass by microbial action at high temperature
journal, January 2013
- Kataeva, Irina; Foston, Marcus B.; Yang, Sung-Jae
- Energy & Environmental Science, Vol. 6, Issue 7
Construction of a Stable Replicating Shuttle Vector for Caldicellulosiruptor Species: Use for Extending Genetic Methodologies to Other Members of This Genus
journal, May 2013
- Chung, Daehwan; Cha, Minseok; Farkas, Joel
- PLoS ONE, Vol. 8, Issue 5, Article No. e62881
Molecular and Biochemical Analyses of CbCel9A/Cel48A, a Highly Secreted Multi-Modular Cellulase by Caldicellulosiruptor bescii during Growth on Crystalline Cellulose
journal, December 2013
- Yi, Zhuolin; Su, Xiaoyun; Revindran, Vanessa
- PLoS ONE, Vol. 8, Issue 12
Interplay between Clostridium thermocellum Family 48 and Family 9 Cellulases in Cellulosomal versus Noncellulosomal States
journal, March 2010
- Vazana, Y.; Morais, S.; Barak, Y.
- Applied and Environmental Microbiology, Vol. 76, Issue 10
Cellulosomes: microbial nanomachines that display plasticity in quaternary structure: Cohesin dockerin recognition
journal, March 2007
- Gilbert, Harry J.
- Molecular Microbiology, Vol. 63, Issue 6
Efficient Degradation of Lignocellulosic Plant Biomass, without Pretreatment, by the Thermophilic Anaerobe "Anaerocellum thermophilum" DSM 6725
journal, May 2009
- Yang, Sung-Jae; Kataeva, Irina; Hamilton-Brehm, Scott D.
- Applied and Environmental Microbiology, Vol. 75, Issue 14, p. 4762-4769
Use of Label-Free Quantitative Proteomics To Distinguish the Secreted Cellulolytic Systems of Caldicellulosiruptor bescii and Caldicellulosiruptor obsidiansis
journal, April 2011
- Lochner, Adriane; Giannone, Richard J.; Rodriguez, Miguel
- Applied and Environmental Microbiology, Vol. 77, Issue 12
The cellulosome — A treasure-trove for biotechnology
journal, September 1994
- Bayer, Edward A.; Morag, Ely; Lamed, Raphael
- Trends in Biotechnology, Vol. 12, Issue 9
Properties and gene structure of a bifunctional cellulolytic enzyme (CelA) from the extreme thermophile 'Anaerocellum thermophilum' with separate glycosyl hydrolase family 9 and 48 catalytic domains
journal, February 1998
- Zverlov, V.; Mahr, S.; Riedel, K.
- Microbiology, Vol. 144, Issue 2, p. 457-465
Dissecting and engineering metabolic and regulatory networks of thermophilic bacteria for biofuel production
journal, November 2013
- Lin, Lu; Xu, Jian
- Biotechnology Advances, Vol. 31, Issue 6
Three Microbial Strategies for Plant Cell Wall Degradation
journal, March 2008
- Wilson, David B.
- Annals of the New York Academy of Sciences, Vol. 1125, Issue 1
Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725
journal, August 2012
- Chung, Daehwan; Farkas, Joel; Huddleston, Jennifer R.
- PLoS ONE, Vol. 7, Issue 8
Improved growth media and culture techniques for genetic analysis and assessment of biomass utilization by Caldicellulosiruptor bescii
journal, November 2012
- Farkas, Joel; Chung, Daehwan; Cha, Minseok
- Journal of Industrial Microbiology & Biotechnology, Vol. 40, Issue 1, p. 41-49
Organization and distribution of the cellulosome in Clostridium thermocellum.
journal, January 1985
- Bayer, E. A.; Setter, E.; Lamed, R.
- Journal of Bacteriology, Vol. 163, Issue 2