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Title: Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases

Abstract

The native recalcitrance of lignocellulosic biomass hinders its effective deconstruction for biological conversion to fuel ethanol. However, once cellulose is physically available to enzymes/microbes, i.e., macro-accessible, cellulose micro-accessibility, i.e., the accessibility as influenced by cellulose properties, further affects cellulose conversion. Here, we performed a comparative study of the effect of cellulose micro-accessibility on cellulose conversion by two biological approaches of potential commercial interest: consolidated bioprocessing (CBP) using Clostridium thermocellum and cell-free saccharification mediated by fungal enzymes. Commercially available cellulosic substrates, Avicel® PH-101, Sigmacell Cellulose Type 50, cotton linters, Whatman™ 1 milled filter paper, and α-cellulose were employed to constitute different cellulose micro-accessibilities. Physiochemical characterization was performed on these substrates to determine key morphological and chemical differences. Biological conversion of these substrates showed that C. was unaffected overall by cellulose structural properties, i.e., micro-accessibility, and achieved similar solids solubilization and metabolite production from these structurally different materials. However, fungal enzymes digested these substrates to different extents. Specifically, glucan conversion of these substrates diminished in the following order: milled filter paper > Avicel > Sigmacell and α-cellulose > cotton linters. Here, we propose that C. thermocellum digestion of lignocellulosic biomass is primarily controlled by the physical availability of cellulose in the lignocellulosicmore » matrix and largely unaffected by cellulose properties once cellulose is made macro-accessible. Finally, in contrast, fungal enzymes require cellulose to be physically accessible, i.e., macro-accessible, as well as have properties amenable to digestion, i.e., micro-accessible.« less

Authors:
ORCiD logo [1]; ORCiD logo [2];  [3]; ORCiD logo [4]; ORCiD logo [5]; ORCiD logo [6];  [7]; ORCiD logo [8]; ORCiD logo [9]; ORCiD logo [10]
  1. Univ. of California, Riverside, CA (United States). Bourns College of Engineering, Dept. of Chemical and Environmental Engineering, Center for Environmental Research and Technology (CE-CERT); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
  2. Univ. of Tennessee, Knoxville, TN (United States). Dept. of Chemical and Biomolecular Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC), and Biosciences Division
  3. Univ. of California, Riverside, CA (United States). Bourns College of Engineering, Dept. of Chemical and Environmental Engineering, Center for Environmental Research and Technology (CE-CERT)
  4. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). UT-ORNL Joint Inst. for Biological Sciences; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
  5. National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center
  6. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). UT-ORNL Joint Inst. for Biological Sciences; 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)
  7. National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center; 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)
  8. Univ. of Tennessee, Knoxville, TN (United States). Dept. of Chemical and Biomolecular Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC), Center for Bioenergy Innovation (CBI),; Univ. of Tennessee, Knoxville, TN (United States). Inst. of Agriculture, Dept. of Forestry, Wildlife, and Fisheries, Center for Renewable Carbon; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Joint Inst. of Biological Sciences, Biosciences Division
  9. 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)
  10. Univ. of California, Riverside, CA (United States). Bourns College of Engineering, Dept. of Chemical and Environmental 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), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1511910
Alternate Identifier(s):
OSTI ID: 1511773
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Green Chemistry
Additional Journal Information:
Journal Name: Green Chemistry; Journal ID: ISSN 1463-9262
Publisher:
Royal Society of Chemistry
Country of Publication:
United States
Language:
English

Citation Formats

Kothari, Ninad, Bhagia, Samarthya, Zaher, Maher, Pu, Yunqiao, Mittal, Ashutosh, Yoo, Chang Geun, Himmel, Michael E., Ragauskas, Arthur J., Kumar, Rajeev, and Wyman, Charles E. Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases. United States: N. p., 2019. Web. doi:10.1039/C9GC00262F.
Kothari, Ninad, Bhagia, Samarthya, Zaher, Maher, Pu, Yunqiao, Mittal, Ashutosh, Yoo, Chang Geun, Himmel, Michael E., Ragauskas, Arthur J., Kumar, Rajeev, & Wyman, Charles E. Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases. United States. doi:10.1039/C9GC00262F.
Kothari, Ninad, Bhagia, Samarthya, Zaher, Maher, Pu, Yunqiao, Mittal, Ashutosh, Yoo, Chang Geun, Himmel, Michael E., Ragauskas, Arthur J., Kumar, Rajeev, and Wyman, Charles E. Wed . "Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases". United States. doi:10.1039/C9GC00262F.
@article{osti_1511910,
title = {Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases},
author = {Kothari, Ninad and Bhagia, Samarthya and Zaher, Maher and Pu, Yunqiao and Mittal, Ashutosh and Yoo, Chang Geun and Himmel, Michael E. and Ragauskas, Arthur J. and Kumar, Rajeev and Wyman, Charles E.},
abstractNote = {The native recalcitrance of lignocellulosic biomass hinders its effective deconstruction for biological conversion to fuel ethanol. However, once cellulose is physically available to enzymes/microbes, i.e., macro-accessible, cellulose micro-accessibility, i.e., the accessibility as influenced by cellulose properties, further affects cellulose conversion. Here, we performed a comparative study of the effect of cellulose micro-accessibility on cellulose conversion by two biological approaches of potential commercial interest: consolidated bioprocessing (CBP) using Clostridium thermocellum and cell-free saccharification mediated by fungal enzymes. Commercially available cellulosic substrates, Avicel® PH-101, Sigmacell Cellulose Type 50, cotton linters, Whatman™ 1 milled filter paper, and α-cellulose were employed to constitute different cellulose micro-accessibilities. Physiochemical characterization was performed on these substrates to determine key morphological and chemical differences. Biological conversion of these substrates showed that C. was unaffected overall by cellulose structural properties, i.e., micro-accessibility, and achieved similar solids solubilization and metabolite production from these structurally different materials. However, fungal enzymes digested these substrates to different extents. Specifically, glucan conversion of these substrates diminished in the following order: milled filter paper > Avicel > Sigmacell and α-cellulose > cotton linters. Here, we propose that C. thermocellum digestion of lignocellulosic biomass is primarily controlled by the physical availability of cellulose in the lignocellulosic matrix and largely unaffected by cellulose properties once cellulose is made macro-accessible. Finally, in contrast, fungal enzymes require cellulose to be physically accessible, i.e., macro-accessible, as well as have properties amenable to digestion, i.e., micro-accessible.},
doi = {10.1039/C9GC00262F},
journal = {Green Chemistry},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {5}
}

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