Probing macromolecular architectures of nanosized cyclic structures of (1-->3)-beta-D-glucans by AFM and SEC-MALLS

Comb-like branched (1-->3)-beta-D-glucans dissolve in water as stiff triple-helical structures. Dissociation followed by re-association leads to the formation of a blend of various macromolecular topologies, where the cyclic species make up a significant fraction. In this study, the molecular properties of these nanosized cyclic structures of (1-->3)-beta-D-glucans were probed using a combination of AFM and SEC-MALLS. The cyclic structures were obtained by subjecting linear triple-helical molecules of (1-->3)-beta-D-glucans to a denaturation-renaturation cycle, and the fraction of cyclic structures in the renatured sample was determined by AFM. Samples containing different known fractions of linear and circular molecules were studied by SEC with online multi-angle laser-light scattering and viscometric detectors. The molecular weight and the radius of gyration of the molecules eluting from the SEC column, as well as the concentration and the intrinsic viscosity, were determined simultaneously. By extrapolating the results to a situation of only circular species, the results allowed to determine the linear mass per unit length (M(L)) of not only the linear but also the circular morphologies of the (1-->3)-beta-D-glucans. The values obtained were M(L)=2140+/-180 g mol(-1)nm(-1) for the circular species and 2045+/-80 g mol(-1)nm(-1) for the linear species. This is the first direct determination of the M(L) parameter of the circular topology, and the results indicate that the reassociation of the individual chains yield a triplex structure also for the circular morphology, similar to the initial triple helix.