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Size Exclusion Chromatography with Multi Detection in Combination with Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry as a Tool for Unraveling the Mechanism of the Enzymatic Polymerization of Polysaccharides

Determination of the size distributions of natural polysaccharides is a challenging task. More advantageous for characterization are well-defined synthetic (hyper)-branched polymers. In this study we concentrated on synthetic amylopectin analogues in order to obtain and compare all available data for different distributions and size dependence of molecular weights. Two groups of well-defined synthetic branched polysaccharides were synthesized via an in vitro enzyme-catalyzed reaction using the enzyme phosphorylase b from rabbit muscle and Deinococcus geothermalis glycogen branching enzyme. Synthetic polymers had a tunable degree of branching (2%–13% determined via 1H NMR) and a tunable degree of polymerization (30–350 determined indirectly via UV spectrometry). The systems used for separation and characterization of branched polysaccharides were SEC-DMSO/LiBr and multi detection (refractive index detector, viscosity detector, and multi angle light scattering detector) and SEC-water/0.02% NaN3; and SEC-50 ...

Authors:   Jelena Ciric; Jorrit Oostland; Jan Willem de Vries; Albert J. J. Woortman; Katja Loos
Journal:   Analytical Chemistry
Year:   2012
DOI:   10.1021/ac302704q
Publication date:   19-Nov-2012
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