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Matrix carbohydrate fractionation and analysis for an MGCx.

ED Eddie Dominguez
RZ Robert Zarnowski
HS Hiram Sanchez
AC Antonio S. Covelli
WW William M. Westler
PA Parastoo Azadi
JN Jeniel Nett
AM Aaron P. Mitchell
DA David R. Andes
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Additional structural analysis was performed after a series of purification and fractionation steps to further ascertain the presence of a mannan-glucan complex. These steps include size exclusion chromatography followed by separation on an anion exchanger HiPrep 16/10 DEAE FF column (GE Healthcare Life Sciences). Neutral free carbohydrates were collected in flowthrough fractions, which were pooled and applied to gel filtration on a HighPrep 16/60 Sephacryl S-300 HR column (GE Healthcare Life Sciences), yielding 22 individual polysaccharide peaks, F1 to F22. The molecular weight of biofilm matrix neutral carbohydrates was determined using size exclusion column calibration with a set of Leuconostoc species dextran standards (polymers with molecular weights [in thousands] of 100, 70, 40, 25, and 6). Both high-molecular-weight (HMW) and low-molecular-weight (LMW) fractions were examined by GC for monosugar analysis. Matrix monosugar composition and quantification was performed on alditol acetate derivatives by GLC-FID (Shimadzu GC-2010 system; Shimadzu, Kyoto, Japan).

Copyright and License information:  ©2018 Dominguez et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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