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Two-Dimensional NMR Evidence for Cleavage of Lignin and Xylan Substituents in Wheat Straw Through Hydrothermal Pretreatment and Enzymatic Hydrolysis

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Abstract

Solution-state two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy of plant cell walls is a powerful tool for characterizing changes in cell wall chemistry during the hydrothermal pretreatment process of wheat straw for second-generation bioethanol production. One-bond 13C–1H NMR correlation spectroscopy, via an heteronuclear single quantum coherence experiment, revealed substantial lignin β-aryl ether cleavage, deacetylation via cleavage of the natural acetates at the 2-O- and 3-O-positions of xylan, and uronic acid depletion via cleavage of the (1 → 2)-linked 4-O-methyl-α-d-glucuronic acid of xylan. In the polysaccharide anomeric region, decreases in the minor β-d-mannopyranosyl, and α-l-arabinofuranosyl units were observed in the NMR spectra from hydrothermally pretreated wheat straw. The aromatic region indicated only minor changes to the aromatic structures during the process (e.g., further deacylation revealed by the depletion in ferulate and p-coumarate structures). Supplementary chemical analyses showed that the hydrothermal pretreatment increased the cellulose and lignin concentration with partial removal of extractives and hemicelluloses. The subsequent enzymatic hydrolysis incurred further deacetylation of the xylan, leaving approximately 10 % of acetate intact based on the weight of original wheat straw.

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Abbreviations

2D NMR:

two-dimensional (solution state) nuclear magnetic resonance spectroscopy

HSQC:

heteronuclear single quantum coherence

β-d-Xylp :

β-d-xylopyranosyl units

2-O-Ac-β-d-Xylp, 3-O-Ac-β-d-Xylp :

O-acetylated β-d-xylopyranosyl units

4-O-MeGlcA:

4-O-methyl-α-d-glucuronic acid units

β-d-Manp :

β-d-mannopyranosyl units

α-l-Araf :

α-l-arabinofuranosyl units

SEM:

scanning electron microscope

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Acknowledgments

The Danish National Advanced Technology Foundation is greatly acknowledged for funding the project “Development of 2nd generation bioethanol process and technology” Project No. 18708. We also gratefully acknowledge the ARS Dairy Forage Research Center, Madison, Wisconsin for use of their NMR spectrometer in the early stages of this research. JR and HK were funded in part by the DOE Great Lakes Bioenergy Research Center (DOE Office of Science BER DE-FC02-07ER64494).

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Authors and Affiliations

  1. U.S. Forest Service, Forest Products Laboratory, Madison, WI, USA

    Daniel J. Yelle, Christopher G. Hunt & Kolby Hirth

  2. Forest & Landscape, Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark

    Prasad Kaparaju & Claus Felby

  3. Department of Biochemistry, DOE Great Lakes Bioenergy Research Center, and Wisconsin Bioenergy Initiative, University of Wisconsin, Madison, WI, USA

    Hoon Kim & John Ralph

  4. Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä, Finland

    Prasad Kaparaju

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  1. Daniel J. Yelle
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Correspondence to Daniel J. Yelle.

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Yelle, D.J., Kaparaju, P., Hunt, C.G. et al. Two-Dimensional NMR Evidence for Cleavage of Lignin and Xylan Substituents in Wheat Straw Through Hydrothermal Pretreatment and Enzymatic Hydrolysis. Bioenerg. Res. 6, 211–221 (2013). https://doi.org/10.1007/s12155-012-9247-6

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