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Title: Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum

Abstract

Background: The thermophilic anaerobic bacterium Clostridium thermocellum is a multifunctional ethanol producer, capable of both saccharifcation and fermentation, that is central to the consolidated bioprocessing (CBP) approach of converting lignocellulosic biomass to ethanol without external enzyme supplementation. Although CBP organisms have evolved efcient machinery for biomass deconstruction, achieving complete solubilization requires targeted approaches, such as pretreatment, to prepare recalcitrant biomass feedstocks for further biological digestion. Here, diferences between how C. thermocellum and fungal cellulases respond to senescent switchgrass prepared by four diferent pretreatment techniques revealed relationships between biomass substrate composition and its digestion by the two biological approaches. Results: Alamo switchgrass was pretreated using hydrothermal, dilute acid, dilute alkali, and co-solvent-enhanced lignocellulosic fractionation (CELF) pretreatments to produce solids with varying glucan, xylan, and lignin compositions. C. thermocellum achieved highest sugar release and metabolite production from de-lignifed switchgrass prepared by CELF and dilute alkali pretreatments demonstrating greater resilience to the presence of hemicellulose sugars than fungal enzymes. 100% glucan solubilization and glucan plus xylan release from switchgrass were achieved using the CELF–CBP combination. Lower glucan solubilization and metabolite production by C. thermocellum was observed on solids prepared by dilute acid and hydrothermal pretreatments with higher xylan removal from switchgrass than ligninmore » removal. Further, C. thermocellum (2% by volume inoculum) showed ~48% glucan solubilization compared to <10% through fungal enzymatic hydrolysis (15 and 65 mg protein/g glucan loadings) of unpretreated switchgrass indicating the efectiveness of C. thermocellum’s cellulosome. Overall, C. thermocellum performed equivalent to 65 and better than 15 mg protein/g glucan fungal enzymatic hydrolysis on all substrates except CELFpretreated substrates. CELF pretreatments of switchgrass produced solids that were highly digestible regardless of whether C. thermocellum or fungal enzymes were chosen. Conclusions: The unparalleled comprehensive nature of this work with a comparison of four pretreatment and two biological digestion techniques provides a strong platform for future integration of pretreatment with CBP. Lignin removal had a more positive impact on biological digestion of switchgrass than xylan removal from the biomass. However, the impact of switchgrass structural properties, including cellulose, hemicellulose, and lignin characterization, would provide a better understanding of lignocellulose deconstruction.« less

Authors:
 [1];  [2];  [3];  [4]; ORCiD logo [5]
  1. Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Dept. of Chemical and Environmental Engineering; Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Center for Environmental Research and Technology (CE-CERT); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
  2. Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
  3. Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Dept. of Chemical and Environmental Engineering; Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Center for Environmental Research and Technology (CE-CERT); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Bioenergy Innovation (CBI)
  4. Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Center for Environmental Research and Technology (CE-CERT); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Bioenergy Innovation (CBI)
  5. Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Dept. of Chemical and Environmental Engineering; Univ. of California, Riverside, CA (United States). Bourns College of Engineering. Center for Environmental Research and Technology (CE-CERT); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Bioenergy Innovation (CBI)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1626995
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 11; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; Biotechnology & Applied Microbiology; Energy & Fuels; Bioethanol; Clostridium thermocellum; Consolidated bioprocessing; Pretreatment; Switchgrass

Citation Formats

Kothari, Ninad, Holwerda, Evert K., Cai, Charles M., Kumar, Rajeev, and Wyman, Charles E. Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum. United States: N. p., 2018. Web. doi:10.1186/s13068-018-1216-7.
Kothari, Ninad, Holwerda, Evert K., Cai, Charles M., Kumar, Rajeev, & Wyman, Charles E. Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum. United States. doi:10.1186/s13068-018-1216-7.
Kothari, Ninad, Holwerda, Evert K., Cai, Charles M., Kumar, Rajeev, and Wyman, Charles E. Sat . "Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum". United States. doi:10.1186/s13068-018-1216-7. https://www.osti.gov/servlets/purl/1626995.
@article{osti_1626995,
title = {Biomass augmentation through thermochemical pretreatments greatly enhances digestion of switchgrass by Clostridium thermocellum},
author = {Kothari, Ninad and Holwerda, Evert K. and Cai, Charles M. and Kumar, Rajeev and Wyman, Charles E.},
abstractNote = {Background: The thermophilic anaerobic bacterium Clostridium thermocellum is a multifunctional ethanol producer, capable of both saccharifcation and fermentation, that is central to the consolidated bioprocessing (CBP) approach of converting lignocellulosic biomass to ethanol without external enzyme supplementation. Although CBP organisms have evolved efcient machinery for biomass deconstruction, achieving complete solubilization requires targeted approaches, such as pretreatment, to prepare recalcitrant biomass feedstocks for further biological digestion. Here, diferences between how C. thermocellum and fungal cellulases respond to senescent switchgrass prepared by four diferent pretreatment techniques revealed relationships between biomass substrate composition and its digestion by the two biological approaches. Results: Alamo switchgrass was pretreated using hydrothermal, dilute acid, dilute alkali, and co-solvent-enhanced lignocellulosic fractionation (CELF) pretreatments to produce solids with varying glucan, xylan, and lignin compositions. C. thermocellum achieved highest sugar release and metabolite production from de-lignifed switchgrass prepared by CELF and dilute alkali pretreatments demonstrating greater resilience to the presence of hemicellulose sugars than fungal enzymes. 100% glucan solubilization and glucan plus xylan release from switchgrass were achieved using the CELF–CBP combination. Lower glucan solubilization and metabolite production by C. thermocellum was observed on solids prepared by dilute acid and hydrothermal pretreatments with higher xylan removal from switchgrass than lignin removal. Further, C. thermocellum (2% by volume inoculum) showed ~48% glucan solubilization compared to <10% through fungal enzymatic hydrolysis (15 and 65 mg protein/g glucan loadings) of unpretreated switchgrass indicating the efectiveness of C. thermocellum’s cellulosome. Overall, C. thermocellum performed equivalent to 65 and better than 15 mg protein/g glucan fungal enzymatic hydrolysis on all substrates except CELFpretreated substrates. CELF pretreatments of switchgrass produced solids that were highly digestible regardless of whether C. thermocellum or fungal enzymes were chosen. Conclusions: The unparalleled comprehensive nature of this work with a comparison of four pretreatment and two biological digestion techniques provides a strong platform for future integration of pretreatment with CBP. Lignin removal had a more positive impact on biological digestion of switchgrass than xylan removal from the biomass. However, the impact of switchgrass structural properties, including cellulose, hemicellulose, and lignin characterization, would provide a better understanding of lignocellulose deconstruction.},
doi = {10.1186/s13068-018-1216-7},
journal = {Biotechnology for Biofuels},
issn = {1754-6834},
number = 1,
volume = 11,
place = {United States},
year = {2018},
month = {8}
}

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