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Title: In situ label-free imaging of hemicellulose in plant cell walls using stimulated Raman scattering microscopy

Plant hemicellulose (largely xylan) is an excellent feedstock for renewable energy production and second only to cellulose in abundance. Beyond a source of fermentable sugars, xylan constitutes a critical polymer in the plant cell wall, where its precise role in wall assembly, maturation, and deconstruction remains primarily hypothetical. Effective detection of xylan, particularly by in situ imaging of xylan in the presence of other biopolymers, would provide critical information for tackling the challenges of understanding the assembly and enhancing the liberation of xylan from plant materials. Raman-based imaging techniques, especially the highly sensitive stimulated Raman scattering (SRS) microscopy, have proven to be valuable tools for label-free imaging. However, due to the complex nature of plant materials, especially those same chemical groups shared between xylan and cellulose, the utility of specific Raman vibrational modes that are unique to xylan have been debated. Here, we report a novel approach based on combining spectroscopic analysis and chemical/enzymatic xylan removal from corn stover cell walls, to make progress in meeting this analytical challenge. We have identified several Raman peaks associated with xylan content in cell walls for label-free in situ imaging xylan in plant cell wall. We demonstrated that xylan can be resolved frommore » cellulose and lignin in situ using enzymatic digestion and label-free SRS microscopy in both 2D and 3D. As a result, we believe that this novel approach can be used to map xylan in plant cell walls and that this ability will enhance our understanding of the role played by xylan in cell wall biosynthesis and deconstruction.« less
Authors:
ORCiD logo [1] ;  [1] ;  [2] ;  [1] ;  [2] ;  [1] ;  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Report Number(s):
NREL/JA-2700-66416
Journal ID: ISSN 1754-6834
Grant/Contract Number:
AC36-08GO28308
Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Research Org:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; hemicellulose; xylan; xylanase; label-free imaging; raman spectroscopy; stimulated Raman scattering (SRS); microscopy
OSTI Identifier:
1335208

Zeng, Yining, Yarbrough, John M., Mittal, Ashutosh, Tucker, Melvin P., Vinzant, Todd B., Decker, Stephen R., and Himmel, Michael E.. In situ label-free imaging of hemicellulose in plant cell walls using stimulated Raman scattering microscopy. United States: N. p., Web. doi:10.1186/s13068-016-0669-9.
Zeng, Yining, Yarbrough, John M., Mittal, Ashutosh, Tucker, Melvin P., Vinzant, Todd B., Decker, Stephen R., & Himmel, Michael E.. In situ label-free imaging of hemicellulose in plant cell walls using stimulated Raman scattering microscopy. United States. doi:10.1186/s13068-016-0669-9.
Zeng, Yining, Yarbrough, John M., Mittal, Ashutosh, Tucker, Melvin P., Vinzant, Todd B., Decker, Stephen R., and Himmel, Michael E.. 2016. "In situ label-free imaging of hemicellulose in plant cell walls using stimulated Raman scattering microscopy". United States. doi:10.1186/s13068-016-0669-9. https://www.osti.gov/servlets/purl/1335208.
@article{osti_1335208,
title = {In situ label-free imaging of hemicellulose in plant cell walls using stimulated Raman scattering microscopy},
author = {Zeng, Yining and Yarbrough, John M. and Mittal, Ashutosh and Tucker, Melvin P. and Vinzant, Todd B. and Decker, Stephen R. and Himmel, Michael E.},
abstractNote = {Plant hemicellulose (largely xylan) is an excellent feedstock for renewable energy production and second only to cellulose in abundance. Beyond a source of fermentable sugars, xylan constitutes a critical polymer in the plant cell wall, where its precise role in wall assembly, maturation, and deconstruction remains primarily hypothetical. Effective detection of xylan, particularly by in situ imaging of xylan in the presence of other biopolymers, would provide critical information for tackling the challenges of understanding the assembly and enhancing the liberation of xylan from plant materials. Raman-based imaging techniques, especially the highly sensitive stimulated Raman scattering (SRS) microscopy, have proven to be valuable tools for label-free imaging. However, due to the complex nature of plant materials, especially those same chemical groups shared between xylan and cellulose, the utility of specific Raman vibrational modes that are unique to xylan have been debated. Here, we report a novel approach based on combining spectroscopic analysis and chemical/enzymatic xylan removal from corn stover cell walls, to make progress in meeting this analytical challenge. We have identified several Raman peaks associated with xylan content in cell walls for label-free in situ imaging xylan in plant cell wall. We demonstrated that xylan can be resolved from cellulose and lignin in situ using enzymatic digestion and label-free SRS microscopy in both 2D and 3D. As a result, we believe that this novel approach can be used to map xylan in plant cell walls and that this ability will enhance our understanding of the role played by xylan in cell wall biosynthesis and deconstruction.},
doi = {10.1186/s13068-016-0669-9},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 9,
place = {United States},
year = {2016},
month = {11}
}

Works referenced in this record:

The preparation of lignocellulosic aerogels from ionic liquid solutions
journal, January 2009

Band assignments in the raman spectra of celluloses
journal, February 1987

Compositional Analysis of Lignocellulosic Feedstocks. 1. Review and Description of Methods
journal, August 2010
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  • Journal of Agricultural and Food Chemistry, Vol. 58, Issue 16, p. 9043-9053
  • DOI: 10.1021/jf1008023