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Data for "Lignin impairs Cel7A degradation of in vitro lignified cellulose by impeding enzyme movement and not by acting as a sink"

Background: Cellulose degradation by cellulases has been studied for decades due to the potential of using lignocellulosic biomass as a sustainable source of bioethanol. In plant cell walls, cellulose is bonded together and strengthened by the polyphenolic polymer, lignin. Because lignin is tightly linked to cellulose and is not digestible by cellulases, is thought to play a dominant role in limiting the efficient enzymatic degradation of plant biomass. Removal of lignin via pretreatments currently limits the cost-efficient production of ethanol from cellulose, motivating the need for a better understanding of how lignin inhibits cellulase-catalyzed degradation of lignocellulose. Work to date using bulk assays has suggested three possible inhibition mechanisms: lignin blocks access of the enzyme to cellulose, lignin impedes progress of the enzyme along cellulose, or lignin binds cellulases directly and acts as a sink.

Results: We used single-molecule fluorescence microscopy to investigate the nanoscale dynamics of Cel7A from Trichoderma reesei, as it binds to and moves along purified bacterial cellulose in vitro. Lignified cellulose was generated by polymerizing coniferyl alcohol onto purified bacterial cellulose, and the degree of lignin incorporation into the cellulose meshwork was analyzed by optical and electron microscopy. We found that Cel7A preferentially bound to regions of cellulose where lignin was absent, and that in regions of high lignin density, Cel7A binding was inhibited. With increasing degrees of lignification, there was a decrease in the fraction of Cel7A that moved along cellulose rather than statically binding. Furthermore, with increasing lignification, the velocity of processive Cel7A movement decreased, as did the distance that individual Cel7A molecules moved during processive runs.

Conclusions: In an in vitro system that mimics lignified cellulose in plant cell walls, lignin did not act as a sink to sequester Cel7A and prevent it from interacting with cellulose. Instead, lignin both blocked access of Cel7A to cellulose and impeded the processive movement of Cel7A along cellulose. This work implies that strategies for improving biofuel production efficiency should target weakening interactions between lignin and cellulose surface, and further suggest that nonspecific adsorption of Cel7A to lignin is likely not a dominant mechanism of inhibition.

Citation

Haviland, Zachary; Nong, Daguan; Zexer, Nerya; Tien, Ming; Anderson, Charles; Hancock, William O. (2024). Data for "Lignin impairs Cel7A degradation of in vitro lignified cellulose by impeding enzyme movement and not by acting as a sink" [Data set]. Scholarsphere. https://doi.org/10.26207/8pe3-rz48

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Metadata

Work Title Data for "Lignin impairs Cel7A degradation of in vitro lignified cellulose by impeding enzyme movement and not by acting as a sink"
Access
Open Access
Creators
  1. Zachary K. Haviland
  2. Daguan Nong
  3. Nerya Zexer
  4. Ming Tien
  5. Charles T. Anderson
  6. William O. Hancock
License CC0 1.0 (Public Domain Dedication)
Work Type Dataset
Publication Date 2024
Language
  1. English
DOI doi:10.26207/8pe3-rz48
Related URLs
  1. https://scholarsphere.psu.edu/resources/9a82f4e6-0c47-483a-a692-503afc1257f4
  2. https://doi.org/10.1186/s13068-023-02456-3
Deposited January 10, 2024

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Work History

Version 1
published

  • Created

    January 10, 2024 11:52 by woh1

  • Updated

    January 10, 2024 11:52 by [unknown user]

  • Added Creator WILLIAM O HANCOCK

    January 10, 2024 11:54 by woh1

  • Added Creator Zachary Haviland

    January 10, 2024 11:54 by woh1

  • Added Creator Daguan Nong

    January 10, 2024 11:54 by woh1

  • Added Creator Nerya Zexer

    January 10, 2024 11:54 by woh1

  • Added Creator Ming Tien

    January 10, 2024 11:54 by woh1

  • Added Creator Charles T Anderson

    January 10, 2024 11:54 by woh1

  • Updated License Show Changes

    January 10, 2024 11:57 by woh1

    License
    • http://creativecommons.org/publicdomain/zero/1.0/
  • Added LigninPaperRevised_Final.pdf

    January 10, 2024 11:59 by woh1

  • Published

    January 10, 2024 11:59 by woh1

  • Updated Publisher Show Changes

    January 11, 2024 14:54 by avs5190

    Publisher
    • Springer Nature
  • Renamed Creator William O. Hancock Show Changes

    January 11, 2024 14:54 by avs5190

    • WILLIAM O HANCOCK
    • William O. Hancock
  • Updated Creator Zachary Haviland

    January 11, 2024 14:54 by avs5190

  • Updated Creator Daguan Nong

    January 11, 2024 14:54 by avs5190

  • Updated Creator Nerya Zexer

    January 11, 2024 14:54 by avs5190

  • Updated Creator Ming Tien

    January 11, 2024 14:54 by avs5190

  • Renamed Creator Charles T. Anderson Show Changes

    January 11, 2024 14:54 by avs5190

    • Charles T Anderson
    • Charles T. Anderson
  • Updated

    April 04, 2024 10:22 by [unknown user]

Version 2
published

  • Created

    January 10, 2024 12:03 by woh1

  • Added T4_full_combine_20211123_0Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T8_full_combine_20210914_1p3Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T1_full_combine_20210914_3Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T7_full_combine_20211017_1p3Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T4_full_combine_20211017_3Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T13_full_combine_20210924_1p9Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T7_full_combine_20210924_1Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T1_full_combine_20210924_9Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T10_full_combine_20211008_0Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T7_full_combine_20211008_1p9Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T4_full_combine_20211008_1Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Added T1_full_combine_20211008_9Lignin.tif

    January 31, 2024 22:12 by zbh5056

  • Published

    January 31, 2024 22:15 by zbh5056

  • Updated

    April 04, 2024 10:22 by [unknown user]

  • Renamed Creator William O. Hancock Show Changes

    February 20, 2025 12:59 by avs5190

    • WILLIAM O HANCOCK
    • William O. Hancock
  • Renamed Creator Zachary K. Haviland Show Changes

    February 20, 2025 12:59 by avs5190

    • Zachary Haviland
    • Zachary K. Haviland
  • Updated Creator Daguan Nong

    February 20, 2025 12:59 by avs5190

  • Updated Creator Nerya Zexer

    February 20, 2025 12:59 by avs5190

  • Updated Creator Ming Tien

    February 20, 2025 12:59 by avs5190

  • Renamed Creator Charles T. Anderson Show Changes

    February 20, 2025 12:59 by avs5190

    • Charles T Anderson
    • Charles T. Anderson

Version 3
published

  • Created

    April 23, 2025 15:36 by woh1

  • Added readme.txt

    April 24, 2025 15:00 by dxn5173

  • Published

    April 24, 2025 15:00 by dxn5173

  • Updated

    April 24, 2025 22:03 by [unknown user]

  • Updated Work Title, Publisher Identifier (DOI) Show Changes

    April 25, 2025 09:41 by avs5190

    Work Title
    • Lignin impairs Cel7A degradation of in vitro lignified cellulose by impeding enzyme movement and not by acting as a sink
    • Data for "Lignin impairs Cel7A degradation of in vitro lignified cellulose by impeding enzyme movement and not by acting as a sink"
    Publisher Identifier (DOI)
    • https://doi.org/10.1186/s13068-023-02456-3
  • Updated Publisher Show Changes

    April 25, 2025 09:41 by avs5190

    Publisher
    • Springer Nature
  • Deleted LigninPaperRevised_Final.pdf

    April 25, 2025 09:42 by avs5190

  • Updated Description Show Changes

    April 25, 2025 09:50 by avs5190

    Description
    • Background: Cellulose degradation by cellulases has been studied for decades due to the potential of using lignocellulosic biomass as a sustainable source of bioethanol. In plant cell walls, cellulose is bonded together and strengthened by the polyphenolic polymer, lignin. Because lignin is tightly linked to cellulose and is not digestible by cellulases, is thought to play a dominant role in limiting the efficient enzymatic degradation of plant biomass. Removal of lignin via pretreatments currently limits the cost-efficient production of ethanol from cellulose, motivating the need for a better understanding of how lignin inhibits cellulase-catalyzed degradation of lignocellulose. Work to date using bulk assays has suggested three possible inhibition mechanisms: lignin blocks access of the enzyme to cellulose, lignin impedes progress of the enzyme along cellulose, or lignin binds cellulases directly and acts as a sink.
    • Results: We used single-molecule fluorescence microscopy to investigate the nanoscale dynamics of Cel7A from Trichoderma reesei, as it binds to and moves along purified bacterial cellulose in vitro. Lignified cellulose was generated by polymerizing coniferyl alcohol onto purified bacterial cellulose, and the degree of lignin incorporation into the cellulose meshwork was analyzed by optical and electron microscopy. We found that Cel7A preferentially bound to regions of cellulose where lignin was absent, and that in regions of high lignin density, Cel7A binding was inhibited. With increasing degrees of lignification, there was a decrease in the fraction of Cel7A that moved along cellulose rather than statically binding. Furthermore, with increasing lignification, the velocity of processive Cel7A movement decreased, as did the distance that individual Cel7A molecules moved during processive runs.
    • Results: We used single-molecule fluorescence microscopy to investigate the nanoscale dynamics of Cel7A from _Trichoderma reesei_, as it binds to and moves along purified bacterial cellulose in vitro. Lignified cellulose was generated by polymerizing coniferyl alcohol onto purified bacterial cellulose, and the degree of lignin incorporation into the cellulose meshwork was analyzed by optical and electron microscopy. We found that Cel7A preferentially bound to regions of cellulose where lignin was absent, and that in regions of high lignin density, Cel7A binding was inhibited. With increasing degrees of lignification, there was a decrease in the fraction of Cel7A that moved along cellulose rather than statically binding. Furthermore, with increasing lignification, the velocity of processive Cel7A movement decreased, as did the distance that individual Cel7A molecules moved during processive runs.
    • Conclusions: In an in vitro system that mimics lignified cellulose in plant cell walls, lignin did not act as a sink to sequester Cel7A and prevent it from interacting with cellulose. Instead, lignin both blocked access of Cel7A to cellulose and impeded the processive movement of Cel7A along cellulose. This work implies that strategies for improving biofuel production efficiency should target weakening interactions between lignin and cellulose surface, and further suggest that nonspecific adsorption of Cel7A to lignin is likely not a dominant mechanism of inhibition.
  • Updated Related URLs Show Changes

    April 25, 2025 09:50 by avs5190

    Related URLs
    • https://scholarsphere.psu.edu/resources/9a82f4e6-0c47-483a-a692-503afc1257f4, https://doi.org/10.1186/s13068-023-02456-3