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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

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. bescii exoproteome in vivo and in vitro. We recently demonstrated that the Acidothermus cellulolyticus E1 endo-1,4-..beta..-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 ..beta..-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:
 [1];  [2];  [2];  [2];  [3]
  1. Univ. of Georgia, Athens, GA (United States); Chung-Ang Univ., Seoul (Korea, Republic of); 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)
  3. Univ. of Georgia, Athens, GA (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 Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1505928
Report Number(s):
NREL/JA-2700-73412
Journal ID: ISSN 1367-5435
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Industrial Microbiology and Biotechnology
Additional Journal Information:
Journal Name: Journal of Industrial Microbiology and Biotechnology; Journal ID: ISSN 1367-5435
Publisher:
Springer
Country of Publication:
United States
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. United States: 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. United States. doi: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". United States. doi:10.1007/s10295-019-02150-0.
@article{osti_1505928,
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. bescii exoproteome in vivo and in vitro. We recently demonstrated that the Acidothermus cellulolyticus E1 endo-1,4-..beta..-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 ..beta..-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 = ,
volume = ,
place = {United States},
year = {2019},
month = {2}
}

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Works referenced in this record:

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

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


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
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