title: Production of bacterial cellulose with controlled deuterium-hydrogen substitution for neutron scattering studies.
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.
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA. Electronic address: oneillhm@ornl.gov.
authors.forename: Hugh
authors.initials: H
authors.lastname: O'Neill
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA; Bredesen Center for Interdisciplinary Research and Graduate Education, University of Tennessee, Knoxville, Tennessee, USA.
authors.forename: Riddhi
authors.initials: R
authors.lastname: Shah
authors.affiliation: Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Barbara R
authors.initials: BR
authors.lastname: Evans
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Junhong
authors.initials: J
authors.lastname: He
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Sai Venkatesh
authors.initials: SV
authors.lastname: Pingali
authors.affiliation: Department of Chemical and Biochemical Engineering, Rutgers University, Piscataway, New Jersey, USA.
authors.forename: Shishir P S
authors.initials: SP
authors.lastname: Chundawat
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan, USA; Department of Chemistry, Michigan State University, East Lansing, Michigan, USA.
authors.forename: A Daniel
authors.initials: AD
authors.lastname: Jones
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Langan
authors.affiliation: Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Brian H
authors.initials: BH
authors.lastname: Davison
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Volker
authors.initials: V
authors.lastname: Urban
mesh_terms: D001419:Bacteria / Q000378:metabolism*; D002482:Cellulose / Q000096:biosynthesis*; D003903:Deuterium / Q000378:metabolism*; D009502:Neutrons; D012542:Scattering, Radiation; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D002482:Cellulose; D003903:Deuterium
keywords: Acetic acid bacteria; Cellulose synthesis; Deuterated growth medium; Deuterium labeling; Deuterium-labeled cellulose; Neutron scattering