We probe the structural and rheological properties of ionically crosslinked alginate, a model biopolymer hydrogel, using microscopy, rheology and viscosity dependent molecular probes. This combination of techniques enables the quantification and correlation of microstructure, microviscosity, bulk viscoelasticity and yielding dynamics. By adjusting the stoichiometric ratio, R, between the alginate biopolymer and its cation crosslinks, we observe a transition from a homogeneous network-like structure to a coarse bundle-like structure at high (R>0.67) stoichiometric ratio. Intriguingly, these bundle-like structures have distinct and counter-intuitive rheological properties. Using molecular probes, we observe a continuous decrease in microviscosity that is correlated with a decrease in bulk elastic modulus and an increase in energy dissipation. This is accompanied by a transition in the hydrogel yielding under strain from a sharp, well-defined yield point to a continuous ductile-like yielding. We ascribe these surprising transitions to the looser intermolecular interaction between alginate biopolymers in the bundle-like state, as previously predicted by x-ray scattering experiments. These findings reveal new and counter-intuitive structure-property relations that demonstrate high crosslink concentration does not necessarily translate to optimal mechanical performance. Significance Alginate is a polysaccharide biopolymer naturally found in brown seaweed cell walls. Extracted alginate forms ionically crosslinked hydrogels that are strong, flexible and increasingly valuable in the biomedical, packaging and food industries. A large part of the utility of these hydrogels stems from the ease with which their mechanics can be tuned through adjusting the stoichiometry between alginate and its ionic crosslinks. However, little is known about how the material properties of alginate hydrogels, in particular their rheology and dynamics, are affected by microscale structural transitions at high stoichiometric ratios. Here, we use a multi-modal approach to describe and correlate hydrogel material properties and to demonstrate that increased polymer crosslinking can, counterintuitively, sometimes weaken hydrogel performance.
Vinay Kopnar, P. Sherin, Sarah P. Graham et al.· bioRxiv· 0 citations
BACKGROUND AND AIMS
The chlorophycean seaweed genus Ulva (sea lettuce and relatives) possesses cellulose and cell-wall-matrix polysaccharides operationally classified as 'pectin-like' (hot-water-extractable anionic ulvans and glucuronans) and hemicelluloses. As Ulva is only distantly related to streptophytes (land-plants plus charophytic algae), their wall polysaccharides are likely to diverge significantly. We therefore explored the little-known structure and properties of Ulva hemicellulose.
METHODS
Hemicellulose-b was alkali-extracted from hot-oxalate-pretreated U. linza alcohol-insoluble residue, then digested with endo-β-xylanase, Driselase or endo-cellulase, yielding several oligosaccharides. These were separated by size on gel-permeation and preparative paper chromatography. Oligosaccharides were dissected by acid hydrolysis, glycosidase digestion and alkaline peeling; products were analysed by thin-layer chromatography. Purified oligosaccharides were also characterised by NMR spectroscopy.
KEY RESULTS
Thirteen di- to pentasaccharides were purified. On acid hydrolysis, they yielded only xylose (Xyl) and/or glucose (Glc). Each was digestible by either β-xylosidase or β-glucosidase, identifying the non-reducing terminus as β-Xyl or β-Glc respectively. The rate of alkaline peeling, which attacks progressively from the reducing terminus, indicated (1→4)-linkages; in each case the smallest peeling product (i.e., originally the non-reducing terminus) was either Xyl or Glc, concurring with the glycosidase digestion result. Structures were deduced [β- (1→4)-linked Xyl-Xyl, Xyl-Xyl-Xyl, Xyl-Glc, Xyl-Xyl-Glc, Xyl-Xyl-Xyl-Glc, Glc-Glc, Glc-Xyl-Xyl, Glc-Glc-Xyl-Xyl, and Xyl-Glc-Glc-Xyl-Xyl], and validated by NMR spectroscopy. Unexpectedly, NMR showed one oligosaccharide to be Glc-3-Glc-4-Glc, the (1→3)-bond probably arising artefactually by trans-β-glucanase activity of the cellulase. Ulva hemicellulose hydrogen-bonded to cellulose more rapidly than any tested land-plant polysaccharide.
CONCLUSIONS
The major hemicellulose of Ulva is a β-(1→4)-linked linear polysaccharide of Xyl and Glc, which we term phycoxyloglucan. The backbone has runs of up to three contiguous β-Xyl and three contiguous β-Glc residues, thus differing fundamentally from land-plant xyloglucan. Its structure confers high affinity for cellulose, suggesting potential biotechnological applications for this seaweed and contributing to the cell wall's mechanical properties.
Marie N Rapin, Lorna Murray, John H. Bothwell et al.· Annals of Botany· 0 citations