Characterization of enzymatically synthesized amylopectin analogs via asymmetrical flow field flow fractionation

•AF4 is a very powerful technique for the analysis of branched polysaccharides.•AF4 was successfully used for fractionation of various branched polysaccharides.•AF4 is especially suited for the characterization of high molecular weight polysaccharides in water.•The mechanism of the enzymatic reaction established via SEC analysis was confirmed.

Asymmetrical flow field flow fractionation (AF4), when coupled with multi-angle laser light scattering (MALLS), is a very powerful technique for determination of the macromolecular structure of high molar mass (branched) polysaccharides. AF4 is a size fractionation technique just as size exclusion chromatography (SEC), nevertheless can overcome some crucial problems found in SEC analysis especially in starch like structures. This paper describes a detailed investigation of the macromolecular structure of two groups of well-defined synthetic amylopectin analogs - synthesized via an in vitro enzyme-catalyzed reaction using the enzymes phosphorylase b from rabbit muscle and Deinococcus geothermalis glycogen branching enzyme (Dg GBE). Size, molar mass distributions and structural data were studied by AF4 coupled with online quasi-elastic light scattering (QELS) and multi-angle light scattering (MALLS).

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