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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

Journal Article · · Journal of Industrial Microbiology and Biotechnology
 [1];  [2];  [2];  [2];  [3]
  1. 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
  2. 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
  3. 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

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.

Research Organization:
National Renewable Energy Laboratory (NREL), Golden, CO (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC05-00OR22725; AC36-08GO28308
OSTI ID:
1772499
Alternate ID(s):
OSTI ID: 1505928
Report Number(s):
NREL/JA-2700-73412
Journal Information:
Journal of Industrial Microbiology and Biotechnology, Journal Name: Journal of Industrial Microbiology and Biotechnology Vol. 46 Journal Issue: 5; ISSN 1367-5435
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
Germany
Language:
English
Citation Metrics:
Cited by: 8 works
Citation information provided by
Web of Science

References (16)

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Engineering the N -terminal end of CelA results in improved performance and growth of Caldicellulosiruptor bescii on crystalline cellulose : Engineering the journal March 2017
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Heterologous expression of a β-d-glucosidase in Caldicellulosiruptor bescii has a surprisingly modest effect on the activity of the exoproteome and growth on crystalline cellulose journal September 2017
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In vivo synergistic activity of a CAZyme cassette from Acidothermus cellulolyticus significantly improves the cellulolytic activity of the C. bescii exoproteome : In Vivo Synergistic Activity of a CAZyme Cassette journal August 2017
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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 journal November 2017
Improved growth media and culture techniques for genetic analysis and assessment of biomass utilization by Caldicellulosiruptor bescii journal November 2012
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Cited By (1)

Genomic and physiological analyses reveal that extremely thermophilic Caldicellulosiruptor changbaiensis deploys uncommon cellulose attachment mechanisms journal August 2019