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Title: Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar

Abstract

Abstract Background Biological conversion of lignocellulosic biomass is significantly hindered by feedstock recalcitrance, which is typically assessed through an enzymatic digestion assay, often preceded by a thermal and/or chemical pretreatment. Here, we assay 17 lines of unpretreated transgenic black cottonwood ( Populus trichocarpa ) utilizing a lignocellulose-degrading, metabolically engineered bacterium, Caldicellulosiruptor bescii . The poplar lines were assessed by incubation with an engineered C. bescii strain that solubilized and converted the hexose and pentose carbohydrates to ethanol and acetate. The resulting fermentation titer and biomass solubilization were then utilized as a measure of biomass recalcitrance and compared to data previously reported on the transgenic poplar samples. Results Of the 17 transgenic poplar lines examined with C. bescii , a wide variation in solubilization and fermentation titer was observed. While the wild type poplar control demonstrated relatively high recalcitrance with a total solubilization of only 20% and a fermentation titer of 7.3 mM, the transgenic lines resulted in solubilization ranging from 15 to 79% and fermentation titers from 6.8 to 29.6 mM. Additionally, a strong inverse correlation ( R 2  = 0.8) between conversion efficiency and lignin content was observed with lower lignin samples more easily converted and solubilized by C. bescii . Conclusionsmore » Feedstock recalcitrance can be significantly reduced with transgenic plants, but finding the correct modification may require a large sample set to identify the most advantageous genetic modifications for the feedstock. Utilizing C. bescii as a screening assay for recalcitrance, poplar lines with down-regulation of coumarate 3-hydroxylase 3 (C3H3) resulted in the highest degrees of solubilization and conversion by C. bescii . One such line, with a growth phenotype similar to the wild-type, generated more than three times the fermentation products of the wild-type poplar control, suggesting that excellent digestibility can be achieved without compromising fitness of the tree.« less

Authors:
; ; ; ; ; ORCiD logo
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1618775
Grant/Contract Number:  
SC0019391
Resource Type:
Published Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Name: Biotechnology for Biofuels Journal Volume: 13 Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
Springer Science + Business Media
Country of Publication:
Netherlands
Language:
English

Citation Formats

Straub, Christopher T., Bing, Ryan G., Wang, Jack P., Chiang, Vincent L., Adams, Michael W. W., and Kelly, Robert M. Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar. Netherlands: N. p., 2020. Web. doi:10.1186/s13068-020-01675-2.
Straub, Christopher T., Bing, Ryan G., Wang, Jack P., Chiang, Vincent L., Adams, Michael W. W., & Kelly, Robert M. Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar. Netherlands. doi:https://doi.org/10.1186/s13068-020-01675-2
Straub, Christopher T., Bing, Ryan G., Wang, Jack P., Chiang, Vincent L., Adams, Michael W. W., and Kelly, Robert M. Wed . "Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar". Netherlands. doi:https://doi.org/10.1186/s13068-020-01675-2.
@article{osti_1618775,
title = {Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar},
author = {Straub, Christopher T. and Bing, Ryan G. and Wang, Jack P. and Chiang, Vincent L. and Adams, Michael W. W. and Kelly, Robert M.},
abstractNote = {Abstract Background Biological conversion of lignocellulosic biomass is significantly hindered by feedstock recalcitrance, which is typically assessed through an enzymatic digestion assay, often preceded by a thermal and/or chemical pretreatment. Here, we assay 17 lines of unpretreated transgenic black cottonwood ( Populus trichocarpa ) utilizing a lignocellulose-degrading, metabolically engineered bacterium, Caldicellulosiruptor bescii . The poplar lines were assessed by incubation with an engineered C. bescii strain that solubilized and converted the hexose and pentose carbohydrates to ethanol and acetate. The resulting fermentation titer and biomass solubilization were then utilized as a measure of biomass recalcitrance and compared to data previously reported on the transgenic poplar samples. Results Of the 17 transgenic poplar lines examined with C. bescii , a wide variation in solubilization and fermentation titer was observed. While the wild type poplar control demonstrated relatively high recalcitrance with a total solubilization of only 20% and a fermentation titer of 7.3 mM, the transgenic lines resulted in solubilization ranging from 15 to 79% and fermentation titers from 6.8 to 29.6 mM. Additionally, a strong inverse correlation ( R 2  = 0.8) between conversion efficiency and lignin content was observed with lower lignin samples more easily converted and solubilized by C. bescii . Conclusions Feedstock recalcitrance can be significantly reduced with transgenic plants, but finding the correct modification may require a large sample set to identify the most advantageous genetic modifications for the feedstock. Utilizing C. bescii as a screening assay for recalcitrance, poplar lines with down-regulation of coumarate 3-hydroxylase 3 (C3H3) resulted in the highest degrees of solubilization and conversion by C. bescii . One such line, with a growth phenotype similar to the wild-type, generated more than three times the fermentation products of the wild-type poplar control, suggesting that excellent digestibility can be achieved without compromising fitness of the tree.},
doi = {10.1186/s13068-020-01675-2},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 13,
place = {Netherlands},
year = {2020},
month = {3}
}

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DOI: https://doi.org/10.1186/s13068-020-01675-2

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Works referenced in this record:

Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees
journal, August 1999

  • Hu, Wen-Jing; Harding, Scott A.; Lung, Jrhau
  • Nature Biotechnology, Vol. 17, Issue 8, p. 808-812
  • DOI: 10.1038/11758

Quantitative fermentation of unpretreated transgenic poplar by Caldicellulosiruptor bescii
journal, August 2019

  • Straub, Christopher T.; Khatibi, Piyum A.; Wang, Jack P.
  • Nature Communications, Vol. 10, Issue 1
  • DOI: 10.1038/s41467-019-11376-6

Current Understanding of the Correlation of Lignin Structure with Biomass Recalcitrance
journal, November 2016


Sugar release and growth of biofuel crops are improved by downregulation of pectin biosynthesis
journal, February 2018

  • Biswal, Ajaya K.; Atmodjo, Melani A.; Li, Mi
  • Nature Biotechnology, Vol. 36, Issue 3
  • DOI: 10.1038/nbt.4067

RNAi-mediated suppression of p-coumaroyl-CoA 3'-hydroxylase in hybrid poplar impacts lignin deposition and soluble secondary metabolism
journal, March 2008

  • Coleman, H. D.; Park, J. -Y.; Nair, R.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 11
  • DOI: 10.1073/pnas.0706537105

Lignin content in natural Populus variants affects sugar release
journal, March 2011

  • Studer, M. H.; DeMartini, J. D.; Davis, M. F.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 15, p. 6300-6305
  • DOI: 10.1073/pnas.1009252108

Lignocellulose conversion: an introduction to chemistry, process and economics
journal, January 2007

  • Lange, Jean-Paul
  • Biofuels, Bioproducts and Biorefining, Vol. 1, Issue 1, p. 39-48
  • DOI: 10.1002/bbb.7

Combinatorial modification of multiple lignin traits in trees through multigene cotransformation
journal, March 2003

  • Li, L.; Zhou, Y.; Cheng, X.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 8, p. 4939-4944
  • DOI: 10.1073/pnas.0831166100

Enhancement of growth and cellulose accumulation by overexpression of xyloglucanase in poplar
journal, March 2004


Antisense Down-Regulation of 4CL Expression Alters Lignification, Tree Growth, and Saccharification Potential of Field-Grown Poplar
journal, August 2010

  • Voelker, Steven L.; Lachenbruch, Barbara; Meinzer, Frederick C.
  • Plant Physiology, Vol. 154, Issue 2
  • DOI: 10.1104/pp.110.159269

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

Engineering redox-balanced ethanol production in the cellulolytic and extremely thermophilic bacterium, Caldicellulosiruptor bescii
journal, December 2018

  • Williams-Rhaesa, Amanda M.; Rubinstein, Gabriel M.; Scott, Israel M.
  • Metabolic Engineering Communications, Vol. 7
  • DOI: 10.1016/j.mec.2018.e00073

Expression of Glycosyl Hydrolases in Lignocellulosic Feedstock: An Alternative for Affordable Cellulosic Ethanol Production
journal, June 2016


Systems and Synthetic Biology of Forest Trees: A Bioengineering Paradigm for Woody Biomass Feedstocks
journal, June 2019

  • Myburg, Alexander A.; Hussey, Steven G.; Wang, Jack P.
  • Frontiers in Plant Science, Vol. 10
  • DOI: 10.3389/fpls.2019.00775

Evaluation of a transgenic poplar as a potential biomass crop for biofuel production
journal, February 2013


Lignin and Biomass: A Negative Correlation for Wood Formation and Lignin Content in Trees
journal, October 2010

  • Novaes, Evandro; Kirst, Matias; Chiang, Vincent
  • Plant Physiology, Vol. 154, Issue 2
  • DOI: 10.1104/pp.110.161281

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

Improving wood properties for wood utilization through multi-omics integration in lignin biosynthesis
journal, April 2018


Significance of Lignin S/G Ratio in Biomass Recalcitrance of Populus trichocarpa Variants for Bioethanol Production
journal, December 2017

  • Yoo, Chang Geun; Dumitrache, Alexandru; Muchero, Wellington
  • ACS Sustainable Chemistry & Engineering, Vol. 6, Issue 2
  • DOI: 10.1021/acssuschemeng.7b03586

Improved saccharification and ethanol yield from field-grown transgenic poplar deficient in cinnamoyl-CoA reductase
journal, December 2013

  • Van Acker, R.; Leple, J. -C.; Aerts, D.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 2
  • DOI: 10.1073/pnas.1321673111

Engineering a thermoregulated intein-modified xylanase into maize for consolidated lignocellulosic biomass processing
journal, October 2012

  • Shen, Binzhang; Sun, Xueguang; Zuo, Xiao
  • Nature Biotechnology, Vol. 30, Issue 11
  • DOI: 10.1038/nbt.2402

New developments in engineering plant metabolic pathways
journal, December 2016


Field-Grown Transgenic Hybrid Poplar with Modified Lignin Biosynthesis to Improve Enzymatic Saccharification Efficiency
journal, February 2017


Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii
journal, June 2017

  • Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.
  • Applied and Environmental Microbiology, Vol. 83, Issue 17
  • DOI: 10.1128/AEM.00969-17

Phylogenetic, Microbiological, and Glycoside Hydrolase Diversities within the Extremely Thermophilic, Plant Biomass-Degrading Genus Caldicellulosiruptor
journal, October 2010

  • Blumer-Schuette, S. E.; Lewis, D. L.; Kelly, R. M.
  • Applied and Environmental Microbiology, Vol. 76, Issue 24, p. 8084-8092
  • DOI: 10.1128/AEM.01400-10

Consolidated bioprocessing of Populus using Clostridium (Ruminiclostridium) thermocellum: a case study on the impact of lignin composition and structure
journal, February 2016

  • Dumitrache, Alexandru; Akinosho, Hannah; Rodriguez, Miguel
  • Biotechnology for Biofuels, Vol. 9, Issue 1
  • DOI: 10.1186/s13068-016-0445-x

Effects on Lignin Structure of Coumarate 3-Hydroxylase Downregulation in Poplar
journal, May 2012


Altered lignin biosynthesis using biotechnology to improve lignocellulosic biofuel feedstocks
journal, July 2014

  • Poovaiah, Charleson R.; Nageswara-Rao, Madhugiri; Soneji, Jaya R.
  • Plant Biotechnology Journal, Vol. 12, Issue 9
  • DOI: 10.1111/pbi.12225

Lignin modification improves the biofuel production potential in transgenic Populus tomentosa
journal, May 2012


An In-Depth Understanding of Biomass Recalcitrance Using Natural Poplar Variants as the Feedstock
journal, December 2016