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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