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Solution-state 2D NMR of Ball-milled Plant Cell Wall Gels in DMSO-d 6

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A Protocol for this article was published on 02 August 2012

Abstract

Although finely divided ball-milled whole cell walls do not completely dissolve in dimethylsulfoxide (DMSO), they readily swell producing a gel. Solution-state two-dimensional (2D) nuclear magnetic resonance (NMR) of this gel, produced directly in the NMR tube, provides an interpretable structural fingerprint of the polysaccharide and lignin components of the wall without actual solubilization, and without structural modification beyond that inflicted by the ball milling and ultrasonication steps. Since the cellulose is highly crystalline and difficult to swell, the component may be under-represented in the spectra. The method however provides a more rapid method for comparative structural evaluation of plant cell walls than is currently available. With the new potential for chemometric analysis using the 2D NMR fingerprint, this method may find application as a secondary screen for selecting biomass lines and for optimizing biomass processing and conversion efficiencies.

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Abbreviations

Ac2O:

acetic anhydride

DMSO:

dimethylsulfoxide

DMSO-d6 :

perdeutero-dimethylsulfoxide

2D:

two-dimensional

FA:

ferulate

G:

guaiacyl

H:

p-hydroxyphenyl

HSQC:

heteronuclear single quantum coherence

HMQC:

heteronuclear multiple quantum coherence

NMR:

nuclear magnetic resonance (spectroscopy)

NMI:

N-methylimidazole

NS:

number of scans

PCA:

p-coumarate

PB:

p-hydroxybenzoate

S:

syringyl

TLC:

thin layer chromatography

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Acknowledgements

We are grateful to Paul Schatz for the locust bean gum sample preparation, to Dan Yelle (U.S. Forest Products Laboratory, Madison, WI) for providing the aspen sample and grinding kenaf bast fiber, and to Fachuang Lu for advice on his cell wall dissolution method. This research was supported by the Office of Science (BER), U.S. Dept. of Energy, Interagency agreement No. DE-AI02-06ER64299. This study made use of 750 and 500 MHz instruments at the National Magnetic Resonance Facility at Madison, which is supported by National Institutes of Health grants P41RR02301 (Biomedical Research Technology Program, National Center for Research Resources) and P41GM66326 (National Institute of General Medical Sciences). Equipment in the facility was purchased with funds from the University of Wisconsin, the National Institutes of Health (P41GM66326, P41RR02301, RR02781, RR08438), the National Science Foundation (DMB-8415048, BIR-9214394), and the U.S. Department of Agriculture.

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Authors and Affiliations

  1. U.S. Dairy Forage Research Center, USDA-Agricultural Research Service, Madison, WI, 53706, USA

    Hoon Kim, John Ralph & Takuya Akiyama

  2. Department of Horticulture, University of Wisconsin, Madison, WI, 53706, USA

    Hoon Kim

  3. Departments of Biochemistry and Biological Systems Engineering, University of Wisconsin, Madison, WI, 53706, USA

    John Ralph

  4. U.S. Dairy Forage Research Center, USDA-ARS, 1925 Linden Drive West, Madison, WI, 53706–1108, USA

    John Ralph

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  1. Hoon Kim
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Correspondence to John Ralph.

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Kim, H., Ralph, J. & Akiyama, T. Solution-state 2D NMR of Ball-milled Plant Cell Wall Gels in DMSO-d 6 . Bioenerg. Res. 1, 56–66 (2008). https://doi.org/10.1007/s12155-008-9004-z

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