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Title: Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose

Abstract

The Caldicellulosiruptor bescii genome encodes a potent set of carbohydrate-active enzymes (CAZymes), found primarily as multi-domain enzymes that exhibit high cellulolytic and hemicellulolytic activity on and allow utilization of a broad range of substrates, including plant biomass without conventional pretreatment. CelA, the most abundant cellulase in the C. bescii secretome, uniquely combines a GH9 endoglucanase and a GH48 exoglucanase in one protein. The most effective commercial enzyme cocktails used in vitro to pretreat biomass are derived from fungal cellulases (cellobiohydrolases, endoglucanases and a β-d-glucosidases) that act synergistically to release sugars for microbial conversion. The C. bescii genome contains six GH5 domains in five different open reading frames. Four exist in multi-domain proteins and two as single catalytic domains. E1 is a GH5 endoglucanase reported to have high specific activity and simple architecture and is active at the growth temperature of C. bescii. E1 is an endo-1,4-β-glucanase linked to a family 2 carbohydrate-binding module shown to bind primarily to cellulosic substrates. As a result, we tested if the addition of this protein to the C. bescii secretome would improve its cellulolytic activity.

Authors:
 [1];  [1];  [1];  [2];  [2];  [2];  [1]
  1. Univ. of Georgia, Athens, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1225358
Report Number(s):
NREL/JA-2700-65047
Journal ID: ISSN 1754-6834
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Related Information: Biotechnology for Biofuels; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANAYLYTICAL CHEMISTRY; crystalline cellulose; acidothermus cellulolyticus e1 endoglucanase

Citation Formats

Chung, Daehwan, Young, Jenna, Cha, Minseok, Brunecky, Roman, Bomble, Yannick J., Himmel, Michael E., and Westpheling, Janet. Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose. United States: N. p., 2015. Web. doi:10.1186/s13068-015-0296-x.
Chung, Daehwan, Young, Jenna, Cha, Minseok, Brunecky, Roman, Bomble, Yannick J., Himmel, Michael E., & Westpheling, Janet. Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose. United States. doi:10.1186/s13068-015-0296-x.
Chung, Daehwan, Young, Jenna, Cha, Minseok, Brunecky, Roman, Bomble, Yannick J., Himmel, Michael E., and Westpheling, Janet. Thu . "Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose". United States. doi:10.1186/s13068-015-0296-x. https://www.osti.gov/servlets/purl/1225358.
@article{osti_1225358,
title = {Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose},
author = {Chung, Daehwan and Young, Jenna and Cha, Minseok and Brunecky, Roman and Bomble, Yannick J. and Himmel, Michael E. and Westpheling, Janet},
abstractNote = {The Caldicellulosiruptor bescii genome encodes a potent set of carbohydrate-active enzymes (CAZymes), found primarily as multi-domain enzymes that exhibit high cellulolytic and hemicellulolytic activity on and allow utilization of a broad range of substrates, including plant biomass without conventional pretreatment. CelA, the most abundant cellulase in the C. bescii secretome, uniquely combines a GH9 endoglucanase and a GH48 exoglucanase in one protein. The most effective commercial enzyme cocktails used in vitro to pretreat biomass are derived from fungal cellulases (cellobiohydrolases, endoglucanases and a β-d-glucosidases) that act synergistically to release sugars for microbial conversion. The C. bescii genome contains six GH5 domains in five different open reading frames. Four exist in multi-domain proteins and two as single catalytic domains. E1 is a GH5 endoglucanase reported to have high specific activity and simple architecture and is active at the growth temperature of C. bescii. E1 is an endo-1,4-β-glucanase linked to a family 2 carbohydrate-binding module shown to bind primarily to cellulosic substrates. As a result, we tested if the addition of this protein to the C. bescii secretome would improve its cellulolytic activity.},
doi = {10.1186/s13068-015-0296-x},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 8,
place = {United States},
year = {2015},
month = {8}
}

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Cited by: 14 works
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Works referenced in this record:

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    Works referencing / citing this record:

    The challenge of enzyme cost in the production of lignocellulosic biofuels
    journal, November 2011

    • Klein-Marcuschamer, Daniel; Oleskowicz-Popiel, Piotr; Simmons, Blake A.
    • Biotechnology and Bioengineering, Vol. 109, Issue 4
    • DOI: 10.1002/bit.24370

    Processive and nonprocessive cellulases for biofuel production—lessons from bacterial genomes and structural analysis
    journal, November 2011


    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
    • DOI: 10.1007/s10295-011-0976-x

    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
    • DOI: 10.1007/s10295-012-1202-1

    Major characteristics of the cellulolytic system of Clostridium thermocellum coincide with those of the purified cellulosome
    journal, January 1985


    The cellulosome — A treasure-trove for biotechnology
    journal, September 1994


    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
    • DOI: 10.1016/j.copbio.2008.04.007

    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
    • DOI: 10.1016/j.jbiosc.2012.07.019

    Designing the deconstruction of plant cell walls
    journal, June 2008


    Crystal Structure of Thermostable Family 5 Endocellulase E1 from Acidothermus cellulolyticus in Complex with Cellotetraose ,
    journal, January 1996

    • Sakon, Joshua; Adney, William S.; Himmel, Michael E.
    • Biochemistry, Vol. 35, Issue 33
    • DOI: 10.1021/bi9604439

    Ultra-Thermostable Cellulases From Acidothermus cellulolyticus: Comparison of Temperature Optima with Previously Reported Cellulases
    journal, August 1989

    • Tucker, Melvin P.; Mohagheghi, Ali; Grohmann, Karel
    • Nature Biotechnology, Vol. 7, Issue 8
    • DOI: 10.1038/nbt0889-817

    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
    • DOI: 10.1039/c3ee40932e

    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
    • DOI: 10.1073/pnas.1402210111

    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
    • DOI: 10.1093/nar/gkq1281

    Isolation and Characterization of Acidothermus cellulolyticus gen. nov., sp. nov., a New Genus of Thermophilic, Acidophilic, Cellulolytic Bacteria
    journal, July 1986

    • MOHAGHEGHI, A.; GROHMANN, K.; HIMMEL, M.
    • International Journal of Systematic Bacteriology, Vol. 36, Issue 3, p. 435-443
    • DOI: 10.1099/00207713-36-3-435

    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
    • DOI: 10.1126/science.1137016

    Revealing Nature's Cellulase Diversity The Digestion Mechanism of Caldicellulosiruptor bescii CelA
    journal, December 2013


    Deletion of Caldicellulosiruptor besciiCelA reveals its crucial role in the deconstruction of lignocellulosic biomass
    journal, October 2014

    • Young, Jenna; Chung, Daehwan; Bomble, Yannick J.
    • Biotechnology for Biofuels, Vol. 7, Issue 1
    • DOI: 10.1186/s13068-014-0142-6

    High activity CAZyme cassette for improving biomass degradation in thermophiles
    journal, February 2018

    • Brunecky, Roman; Chung, Daehwan; Sarai, Nicholas S.
    • Biotechnology for Biofuels, Vol. 11, Issue 1
    • DOI: 10.1186/s13068-018-1014-2

    Three Microbial Strategies for Plant Cell Wall Degradation
    journal, March 2008


    Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of Caldicellulosiruptor bescii DSM6725
    journal, August 2012


    Homologous Expression of the Caldicellulosiruptor bescii CelA Reveals that the Extracellular Protein Is Glycosylated
    journal, March 2015


    Synergistic interactions in cellulose hydrolysis
    journal, January 2012


    High activity CAZyme cassette for improving biomass degradation in thermophiles
    journal, February 2018

    • Brunecky, Roman; Chung, Daehwan; Sarai, Nicholas S.
    • Biotechnology for Biofuels, Vol. 11, Issue 1
    • DOI: 10.1186/s13068-018-1014-2