Abstract
Isotopic enrichment of biomacromolecules is a widely used technique that enables the investigation of the structural and dynamic properties to provide information not accessible with natural abundance isotopic composition. This study reports an approach for deuterium incorporation into bacterial cellulose. A media formulation for growth of Acetobacter xylinus subsp. sucrofermentans and Gluconacetobacter hansenii was formulated that supports cellulose production in deuterium (D) oxide. The level of D incorporation can be varied by altering the ratio of deuterated and protiated glycerol used during cell growth in the D2O-based growth medium. Spectroscopic analysis and mass spectrometry show that the level of deuterium incorporation is high (> 90%) for the perdeuterated form of bacterial cellulose. The small-angle neutron scattering profiles of the cellulose with different amounts of D incorporation are all similar indicating that there are no structural changes in the cellulose due to substitution of deuterium for hydrogen. In addition, by varying the amount of deuterated glycerol in the media it was possible to vary the scattering length density of the deuterated cellulose. The ability to control deuterium content of cellulose extends the range of experiments using techniques such as neutron scattering to reveal information about the structure and dynamics of cellulose, and its interactions with other biomacromolecules as well as synthetic polymers used for development of composite materials.
Original language | English |
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Title of host publication | Methods in Enzymology |
Publisher | Academic Press Inc. |
Pages | 123-146 |
Number of pages | 24 |
DOIs | |
State | Published - 2015 |
Publication series
Name | Methods in Enzymology |
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Volume | 565 |
ISSN (Print) | 0076-6879 |
ISSN (Electronic) | 1557-7988 |
Funding
This manuscript has been authored by UT-Battelle, LLC under Contract No. DE-AC05-00OR22725 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a nonexclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan ( http://energy.gov/downloads/doe-public-access-plan ). This research is funded by the Genomic Science Program, Office of Biological and Environmental Research, U.S. Department of Energy, under FWP ERKP752. The Center for Structural Molecular Biology (Project ERKP291) and the Bio-SANS beam line is supported by the Office of Biological and Environmental Research U.S. Department of Energy. Research at the Spallation Neutron Source and High Flux Isotope Reactor at Oak Ridge National Laboratory (ORNL) is supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, U.S. Department of Energy (DOE). ORNL is managed by UT-Battelle, LLC, for the U.S. DOE under Contract No. DE-AC05-00OR22725.