skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously

Abstract

Here, cellulose and hemicellulose are the most abundant components in plant biomass. A preferred Consolidated Bioprocessing (CBP) system is one which can directly convert both cellulose and hemicellulose into target products without adding the costly hydrolytic enzyme cocktail. In this work, the thermophilic, cellulolytic, and anaerobic bacterium, Clostridium thermocellum DSM 1313, was engineered to grow on xylose in addition to cellulose. Both xylA (encoding for xylose isomerase) and xylB (encoding for xylulokinase) genes from the thermophilic anaerobic bacterium Thermoanaerobacter ethanolicus were introduced to enable xylose utilization while still retaining its inherent ability to grow on 6-carbon substrates. Targeted integration of xylAB into C. thermocellum genome realized simultaneous fermentation of xylose with glucose, with cellobiose (glucose dimer), and with cellulose, respectively, without carbon catabolite repression. We also showed that the respective H 2 and ethanol production were twice as much when both xylose and cellulose were consumed simultaneously than when consuming cellulose alone. Moreover, the engineered xylose consumer can also utilize xylo-oligomers (with degree of polymerization of 2-7) in the presence of xylose. Isotopic tracer studies also revealed that the engineered xylose catabolism contributed to the production of ethanol from xylan which is a model hemicellulose in mixed sugar fermentation, demonstratingmore » immense potential of this enhanced CBP strain in co-utilizing both cellulose and hemicellulose for the production of fuels and chemicals.« less

Authors:
 [1];  [2];  [1];  [1]; ORCiD logo [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States); Univ. de los Andes, Bogota (Columbia)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Hydrogen and Fuel Cell Technologies Program (EE-3F); USDOE Office of Energy Efficiency and Renewable Energy (EERE), NREL Laboratory Directed Research and Development (LDRD); USDOE
OSTI Identifier:
1430817
Alternate Identifier(s):
OSTI ID: 1432725
Report Number(s):
NREL/JA-2700-71150
Journal ID: ISSN 0006-3592
Grant/Contract Number:  
AC36-08GO28308; 0627-1403
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology and Bioengineering
Additional Journal Information:
Journal Volume: 115; Journal Issue: 7; Journal ID: ISSN 0006-3592
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; lignocellulose; Clostridium thermocellum; consolidated bioprocessing (CBP); xylose; biohydrogen; thermophile

Citation Formats

Xiong, Wei, Reyes, Luis H., Michener, William E., Maness, Pin -Ching, and Chou, Katherine J. Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously. United States: N. p., 2018. Web. doi:10.1002/bit.26590.
Xiong, Wei, Reyes, Luis H., Michener, William E., Maness, Pin -Ching, & Chou, Katherine J. Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously. United States. doi:10.1002/bit.26590.
Xiong, Wei, Reyes, Luis H., Michener, William E., Maness, Pin -Ching, and Chou, Katherine J. Tue . "Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously". United States. doi:10.1002/bit.26590. https://www.osti.gov/servlets/purl/1430817.
@article{osti_1430817,
title = {Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously},
author = {Xiong, Wei and Reyes, Luis H. and Michener, William E. and Maness, Pin -Ching and Chou, Katherine J.},
abstractNote = {Here, cellulose and hemicellulose are the most abundant components in plant biomass. A preferred Consolidated Bioprocessing (CBP) system is one which can directly convert both cellulose and hemicellulose into target products without adding the costly hydrolytic enzyme cocktail. In this work, the thermophilic, cellulolytic, and anaerobic bacterium, Clostridium thermocellum DSM 1313, was engineered to grow on xylose in addition to cellulose. Both xylA (encoding for xylose isomerase) and xylB (encoding for xylulokinase) genes from the thermophilic anaerobic bacterium Thermoanaerobacter ethanolicus were introduced to enable xylose utilization while still retaining its inherent ability to grow on 6-carbon substrates. Targeted integration of xylAB into C. thermocellum genome realized simultaneous fermentation of xylose with glucose, with cellobiose (glucose dimer), and with cellulose, respectively, without carbon catabolite repression. We also showed that the respective H2 and ethanol production were twice as much when both xylose and cellulose were consumed simultaneously than when consuming cellulose alone. Moreover, the engineered xylose consumer can also utilize xylo-oligomers (with degree of polymerization of 2-7) in the presence of xylose. Isotopic tracer studies also revealed that the engineered xylose catabolism contributed to the production of ethanol from xylan which is a model hemicellulose in mixed sugar fermentation, demonstrating immense potential of this enhanced CBP strain in co-utilizing both cellulose and hemicellulose for the production of fuels and chemicals.},
doi = {10.1002/bit.26590},
journal = {Biotechnology and Bioengineering},
number = 7,
volume = 115,
place = {United States},
year = {2018},
month = {4}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 11 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

The Cellulosomes: Multienzyme Machines for Degradation of Plant Cell Wall Polysaccharides
journal, October 2004


Engineering the N -terminal end of CelA results in improved performance and growth of Caldicellulosiruptor bescii on crystalline cellulose : Engineering the
journal, March 2017

  • Kim, Sun-Ki; Chung, Daehwan; Himmel, Michael E.
  • Biotechnology and Bioengineering, Vol. 114, Issue 5
  • DOI: 10.1002/bit.26242

Dilute-Sulfuric Acid Pretreatment of Corn Stover in Pilot-Scale Reactor: Investigation of Yields, Kinetics, and Enzymatic Digestibilities of Solids
journal, January 2003

  • Schell, Daniel J.; Farmer, Jody; Newman, Millie
  • Applied Biochemistry and Biotechnology, Vol. 105, Issue 1-3, p. 69-86
  • DOI: 10.1385/ABAB:105:1-3:69

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
  • DOI: 10.1073/pnas.1003584107

Cellulosomes: plant-cell-wall-degrading enzyme complexes
journal, July 2004

  • Doi, Roy H.; Kosugi, Akihiko
  • Nature Reviews Microbiology, Vol. 2, Issue 7
  • DOI: 10.1038/nrmicro925

Microbial Cellulose Utilization: Fundamentals and Biotechnology
journal, September 2002

  • Lynd, L. R.; Weimer, P. J.; van Zyl, W. H.
  • Microbiology and Molecular Biology Reviews, Vol. 66, Issue 3, p. 506-577
  • DOI: 10.1128/MMBR.66.3.506-577.2002

Induction of lignocellulose-degrading enzymes in Neurospora crassa by cellodextrins
journal, April 2012

  • Znameroski, E. A.; Coradetti, S. T.; Roche, C. M.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 16
  • DOI: 10.1073/pnas.1118440109

Enzymatic hydrolysis of cellulosic biomass
journal, July 2011


CO 2 -fixing one-carbon metabolism in a cellulose-degrading bacterium Clostridium thermocellum
journal, October 2016

  • Xiong, Wei; Lin, Paul P.; Magnusson, Lauren
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 46
  • DOI: 10.1073/pnas.1605482113

Cellulase, Clostridia, and Ethanol
journal, March 2005

  • Demain, A. L.; Newcomb, M.; Wu, J. H. D.
  • Microbiology and Molecular Biology Reviews, Vol. 69, Issue 1, p. 124-154
  • DOI: 10.1128/MMBR.69.1.124-154.2005

Carbon catabolite repression in Thermoanaerobacterium saccharolyticum
journal, January 2012

  • Tsakraklides, Vasiliki; Shaw, A.; Miller, Bethany B.
  • Biotechnology for Biofuels, Vol. 5, Issue 1
  • DOI: 10.1186/1754-6834-5-85

Pyruvate catabolism and hydrogen synthesis pathway genes of Clostridium thermocellum ATCC 27405
journal, June 2008

  • Carere, Carlo R.; Kalia, Vipin; Sparling, Richard
  • Indian Journal of Microbiology, Vol. 48, Issue 2
  • DOI: 10.1007/s12088-008-0036-z

Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production
journal, April 2016

  • Liu, Zhuo; Ho, Shih-Hsin; Sasaki, Kengo
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep24550

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

Carbon catabolite repression in bacteria: many ways to make the most out of nutrients
journal, August 2008

  • Görke, Boris; Stülke, Jörg
  • Nature Reviews Microbiology, Vol. 6, Issue 8, p. 613-624
  • DOI: 10.1038/nrmicro1932

Engineered Saccharomyces cerevisiae capable of simultaneous cellobiose and xylose fermentation
journal, December 2010

  • Ha, S. -J.; Galazka, J. M.; Rin Kim, S.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 2
  • DOI: 10.1073/pnas.1010456108

Metabolic Engineering of Clostridium cellulolyticum for Production of Isobutanol from Cellulose
journal, March 2011

  • Higashide, Wendy; Li, Yongchao; Yang, Yunfeng
  • Applied and Environmental Microbiology, Vol. 77, Issue 8
  • DOI: 10.1128/AEM.02454-10

Biohydrogen as a renewable energy resource—Prospects and potentials
journal, January 2008


Clostridium thermocellum cellulosomal genes are regulated by extracytoplasmic polysaccharides via alternative sigma factors
journal, October 2010

  • Nataf, Y.; Bahari, L.; Kahel-Raifer, H.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 43
  • DOI: 10.1073/pnas.1012175107

Continuous hydrogen production during fermentation of α-cellulose by the thermophillic bacterium Clostridium thermocellum
journal, February 2009

  • Magnusson, Lauren; Cicek, Nazim; Sparling, Richard
  • Biotechnology and Bioengineering, Vol. 102, Issue 3
  • DOI: 10.1002/bit.22092

Biocommodity Engineering
journal, October 1999

  • Lynd, L. R.; Wyman, C. E.; Gerngross, T. U.
  • Biotechnology Progress, Vol. 15, Issue 5
  • DOI: 10.1021/bp990109e

Cellodextrin and Laminaribiose ABC Transporters in Clostridium thermocellum
journal, October 2008

  • Nataf, Y.; Yaron, S.; Stahl, F.
  • Journal of Bacteriology, Vol. 191, Issue 1
  • DOI: 10.1128/JB.01190-08

How biotech can transform biofuels
journal, February 2008

  • Lynd, Lee R.; Laser, Mark S.; Bransby, David
  • Nature Biotechnology, Vol. 26, Issue 2, p. 169-172
  • DOI: 10.1038/nbt0208-169

The cellulosome and cellulose degradation by anaerobic bacteria
journal, September 2001


Cellulose utilization by Clostridium thermocellum: Bioenergetics and hydrolysis product assimilation
journal, May 2005

  • Zhang, Y. -H. P.; Lynd, L. R.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 20
  • DOI: 10.1073/pnas.0408734102

Relationship of cellulosomal and noncellulosomal xylanases of Clostridium thermocellum to cellulose-degrading enzymes.
journal, October 1990


Engineered Escherichia coli capable of co-utilization of cellobiose and xylose
journal, January 2012


High Ethanol Titers from Cellulose by Using Metabolically Engineered Thermophilic, Anaerobic Microbes
journal, September 2011

  • Argyros, D. Aaron; Tripathi, Shital A.; Barrett, Trisha F.
  • Applied and Environmental Microbiology, Vol. 77, Issue 23, p. 8288-8294
  • DOI: 10.1128/AEM.00646-11

Hydrogen production from cellulose in a two-stage process combining fermentation and electrohydrogenesis
journal, August 2009

  • Lalaurette, Elodie; Thammannagowda, Shivegowda; Mohagheghi, Ali
  • International Journal of Hydrogen Energy, Vol. 34, Issue 15
  • DOI: 10.1016/j.ijhydene.2009.05.112