Variation of xyloglucan substitution pattern affects the sorption on celluloses with different degrees of crystallinity

The sorption of xyloglucan (XG) on cellulose is a basic feature of the supramolecular assembly of plant cell walls. The binding to cellulose of xyloglucan fractions from Rubus fruticosus suspension-cultured cells with different substitution patterns was assayed on celluloses having various degrees of crystallinity between 20 and 95%. The primary structure of XGs differing in their Xyl/Glc ratio affected their binding to cellulose. The less substituted XGs gave the highest binding yields. Selective removal of the terminal fucosyl residues of XGs differentially affected the binding depending on the crystallinity of cellulose. The results showed large variations on the way cellulose crystallinity affects the binding interaction of XGs. Interestingly, one of the highest binding capacities was exhibited by the primary cell wall cellulose isolated from the actual R. fruticosus cells which also had the lowest crystallinity. Differences in binding to primary wall cellulose appeared to be inversely related to the global substitution of the glucan main chain of XGs.     

Structural Characteristics of Xyloglucan – Congo Red Aggregates as Observed by Small Angle X-ray Scattering

Xyloglucan is a type of hemicellulose with a cellulose backbone containing (1→6)-α-xylose or (1→2)-β-galactoxylose as a side chain. It is soluble in water. Its aqueous solution forms a gel or gel-like precipitate by addition of Congo red. Xyloglucan gel structures with various concentrations of Congo red were observed by small angle X-ray scattering (SAXS) at the nano-level. SAXS results indicated that the xyloglucan chains interacted with Congo red, and that an increase of concentration of Congo red induced a characteristic cross-linking domain, which consisted of a flat structure containing stacked xyloglucan chain assemblies. The Congo red molecules are inserted between the xyloglucan chains.     

Gelation of xyloglucan ID water/alcohol systems

Xyloglucan has a cellulose backbone with branched (16)--xylose or (12)--galactoxylose as a side chain. Its aqueous solution yields a gel by adding alcohol. The gel structure of xyloglucan ID various kinds of mono- or polyhydric alcohol/water systems was studied by small-angle X-ray scattering (SAXS). The gelation behavior ID strongly dependent on the type of alcohol. The SAXS from gel with monohydric alcohols indicated that the xyloglucan chains caused random aggregation, as expressed with a Debye–Bueche type scattering function. The type of alcohol added was correlated with the size of the inhomogeneity, as evaluated by SAXS results. The gelation with polyhydric alcohols resulted ID less association, which occurred as side-by-side association with a few xyloglucan chains, rather than as random aggregation.     

Roles of xyloglucan and pectin on the mechanical properties of bacterial cellulose composite films

Xyloglucan and pectin are major non-cellulosic components of most primary plant cell walls. It is believed that xyloglucan and perhaps pectin are functioning as tethers between cellulose microfibrils in the cell walls. In order to understand the role of xyloglucan and pectin in cell wall mechanical properties, model cell wall composites created using Gluconacetobacter xylinus cellulose or cellulose nanowhiskers (CNWs) derived there from with different amounts of xyloglucan and/or pectin have been prepared and measured under extension conditions. Compared with pure CNW films, CNW composites with lower amounts of xyloglucan or pectin did not show significant differences in mechanical behavior. Only when the additives were as high as 60 %, the films exhibited a slightly lower Young’s modulus. However, when cultured with xyloglucan or pectin, the bacterial cellulose (BC) composites produced by G. xylinus showed much lower modulus compared with that of the pure BC films. Xyloglucan was able to further reduce the modulus and extensibility of the film compared to that of pectin. It is proposed that surface coating or tethering of xyloglucan or pectin of cellulose microfibrils does not alone affect the mechanical properties of cell wall materials. The implication from this work is that xyloglucan or pectin alters the mechanical properties of cellulose networks during rather than after the cellulose biosynthesis process, which impacts the nature of the connection between these compounds.     

Solvation of xyloglucan in water/alcohol systems by molecular dynamics simulation

Xyloglucan in water solution turns into a gel with addition of alcohol such as methanol and ethanol. In regard to this phenomenon, we investigated the adhesive property of alcohol to xyloglucan and proposed the mechanism of the gelation by molecular dynamics (MD) simulation of a xyloglucan in water, water/methanol, and water/ethanol solution for 10 ns. The alcohol molecules showed its adhesive property to the xyloglucan and made the swelling-shrinking motion of the xyloglucan slow. Alcohol molecules solvated to the xyloglucan mainly in hydrophobic way so as to fill the void of water hydration shell, resulting in reformation of the hydrogen-bond network of water molecules around the solute. We also found that alcohol molecules have strong tendency to hydrogen-bond on xylose O3 in xyloglucan. According to these results, we proposed the gelation mechanism of xyloglucan in water/alcohol solution.     

The effect of sterilization methods on the thermo-gelation properties of xyloglucan hydrogels

In this study, the influence of different sterilization methods on the thermo-gelation and structural properties of xyloglucan hydrogels was investigated. Xyloglucan samples were treated by either 70% ethanol, 70% isopropanol, γ-irradiation (10 kGy) at room temperature, γ-irradiation (10 kGy) in dry ice or autoclaving. These samples were tested for sterility by incubation with sterile Lysogeny Broth (LB) at room temperature, 30 °C and 37 °C for 30 days. According to the results obtained, xyloglucan hydrogels were only effectively sterilized by autoclaving or by γ-irradiation either at room temperature or in dry ice. These samples were analyzed by rheology measurements and dynamic and static light scattering analysis. Gamma-irradiation at room temperature markedly changed the polymer structure, preventing thermo-gelation. Only autoclaving and γ-irradiation in dry ice preserved the rheological properties of the polymer. The sol-gel transition as a function of the temperature was similar for these samples and the control sample.     

Characterizing wood polymers in the primary cell wall of Norway spruce (<Emphasis Type="Italic">Picea abies</Emphasis> (L.) Karst.) using dynamic FT-IR spectroscopy

Dynamic Fourier Transform Infra-Red (FT-IR) spectroscopy was used to examine the interactions among cellulose, xyloglucan, pectin, protein and lignin in the outer fibre wall layers of spruce wood tracheids. Knowledge regarding these interactions is fundamental for understanding the fibre separation in a mechanical pulping process. Sheets made from an enriched primary cell wall material were used for studying the viscoelastic response of the polymers. The results indicated that strong interactions exist among lignin, protein, pectin, xyloglucan and cellulose in the primary cell wall. This signified a closely linked network structure of the components on the fibre surface. This ultrastructural arrangement in the primary cell wall and the relatively high content of lignin, pectin and protein in it, means that the primary cell wall is more submissive to selective chemical attacks, when compared to the secondary cell wall. A low ratio of cellulose Iα to cellulose Iβ in the primary cell wall was also found.     

Rheological and related study of gelation of xyloglucan in the presence of small molecules and other polysaccharides

Interaction between tamarind seed xyloglucan and the other polysaccharides, gellan gum or xanthan investigated by rheology, differential scanning calorimetry, and related methods was discussed. All these three polysaccharides do not form a gel at lower concentrations by itself at the experimental conditions studied but the gelation of xyloglucan occurs in the presence of gellan or xanthan. Gelation of xyloglucan in the presence of a polyphenol, epigallocatechin gallate, is also discussed. Hence the gelation of these mixtures is caused by the synergistic interaction, and the models for the synergistic interaction were discussed. The gelation of polysaccharides by the synergistic interaction is of great value for food and related industries.     

Revision of adsorption models of xyloglucan on microcrystalline cellulose

Interactions among cellulose, hemicellulose and pectins are important for plant cell wall assembly and properties and also for industrial applications of these polysaccharides. Therefore, binding of pectin and xyloglucan on microcrystalline cellulose was investigated in this experiment by adsorption isotherms, zeta potential and scanning electron microscopy (SEM). Analysis of three isotherm models (Langmuir, Freundlich and Fowler-Guggenheim isotherms) showed that the experimental adsorption isotherm was well described via the Fowler-Guggenheim model, which includes lateral interaction between the adsorbate. The adsorption isotherm and zeta potential measurement showed that at temperature 25 °C only xyloglucan adsorbed on the microcrystalline cellulose. In case of xyloglucan on cellulose, the equilibrium was reached in about 3–4 h, and the kinetics of adsorption were well described by the multiexponential equation. Analysis of the model suggests that two steps can be distinguished: diffusion and reconformation in an adsorbed layer. No adsorption of pectin was observed in this study. SEM study showed that xyloglucan may prevent cellulose from aggregation.     

The impact of cellulose structure on binding interactions with hemicellulose and pectin

Four cellulose substrates including highly crystalline cellulose nanowhiskers (CNWs) from Gluconacetobacter xylinus (cellulose Iα) or cotton (cellulose Iβ) and amorphous cellulose derived from CNWs (phosphoric acid swollen cellulose nanowhiskers, PASCNWs) were used to explore the interaction between cellulose and well-defined xyloglucan, xylan, arabinogalactan and pectin. The binding behavior was characterized by adsorption isotherm and Langmuir models. The maximum adsorption and the binding constant of xyloglucan, xylan and pectin to any CNWs were always higher than to PASCNWs derived from the same source. The binding affinity of xyloglucan, xylan and pectin to G. xylinus cellulose was generally higher than to cotton cellulose, showing that binding interactions depended on the biological origin of cellulose and associated differences in its structure. The surface area, porosity, crystal plane and degree of order of cellulose substrate may all impact the interactions.     

Models of Xyloglucan Binding to Cellulose Microfibrils1

Abstract

Molecular modeling is used to investigate the ways in which plant cell wall xyloglucans might bind to the surface of cellulose microfibrils. Binding involving the xyloglucan backbone is found to be sterically restricted. Plausible models are obtained that involve hydrogen bonding between the xylose residues and one kind of cellulose surface. In such a model, the xyloglucan sidechains mediate, as well as modulate, the binding.     

Activation of crystalline cellulose surfaces through the chemoenzymatic modification of xyloglucan

Cellulose constitutes an important raw material for many industries. However, the superb load-bearing properties of cellulose are accompanied by poor chemical reactivity. The hydroxyl groups on cellulose surfaces can be reacted but usually not without loss of fiber integrity and strength. Here, we describe a novel chemoenzymatic approach for the efficient incorporation of chemical functionality onto cellulose surfaces. The modification is brought about by using a transglycosylating enzyme, xyloglucan endotranglycosylase, to join chemically modified xyloglucan oligosaccharides to xyloglucan, which has a naturally high affinity to cellulose. Binding of the chemically modified hemicellulose molecules can thus be used to attach a wide variety of chemical moieties without disruption of the individual fiber or fiber matrix.     

Non-electrostatic building of biomimetic cellulose-xyloglucan multilayers

Layer-by-layer assembly was used to build thin films, consisting of multiple layers alternating cellulose nanocrystals and xyloglucan, benefiting from the strong non-electrostatic cellulose-xyloglucan interaction. Data from atomic force microscopy and neutron reflectivity showed that these well-defined films exhibited a thickness increasing linearly with the number of layers, without increase in surface roughness. These "green" nanocomposite films, reminiscent of plant cell wall, are composed of a regular stack of single layers of cellulose nanocrystals separated by very thin xyloglucan spacers. Such architecture differs from the one formed by cellulose/polycations multilayers, where the cellulose phase itself consists of a double layer.     

Preparation of Arabinoxylan and its Sorption on Bacterial Cellulose During Cultivation

The sorption of arabinoxylan (AX) on bacterial cellulose was investigated by adding AX to the culture medium of Gluconacetobacter xylinus. The starting AX material was produced by alkaline extraction of oat spelts. To investigate the impact of varying AX quality, the residual lignin was reduced by ClO₂ bleaching. Furthermore, bleached and unbleached xylans were subjected to xylanase hydrolysis in order to produce fractions of varying molar mass. Of all samples only the water soluble fractions were used for sorption experiments. A reduced molar mass resulted in a lower sorption of AX to the cellulose, while the lignin content increased the sorption of AX on bacterial cellulose. The sorption of AX resulted in a reduction of bacterial cellulose crystallinity and cellulose Iα content. In combined treatments of AX with xyloglucan and β-glucan no synergistic effect of those polysaccharides on the AX sorption was found.     

Incorporation of β-(1,6)-linked glucooligosaccharides (pustulooligosaccharides) into plant cell wall structures

Protein extract of germinating nasturtium (Tropaeolum majus) seeds containing xyloglucan endotransglycosylase (xyloglucan xyloglucosyl transferase, EC 2.4.1.207, abbreviated XET) exhibited the heterotransglycosylating activity with donor/acceptor substrate pair xyloglucan/sulphorhodamine labelled pustulooligosaccharides (XG/PUOS-SR) in a dot blot assay. The heterotransglycosylating activity was confirmed by the substrate-product changes during transglycosylation by HPLC size-exclusion chromatography. Another donor substrate capable of being coupled with PUOS-SR was cellulose, probably owing to its structural similarity to xyloglucan. Surprisingly, microscopic comparison of the incorporation of the labelled xyloglucan nonasaccharide XGO9-SR (specific substrate for XET) and PUOS-SR into the cell wall structures clearly showed differences in their binding to specific cell structures: the primary cell wall and the plasma membrane. These findings indicate the existence in nasturtium of XETs with different localisation, substrate specificity and, probably, function.     

Structure characterization of native cellulose during dehydration and rehydration

The goal of this study is to investigate the hydration and dehydration induced structural changes of native cellulose. Never dried cotton, and never dried bacterial cellulose with and without added matrix polymer xyloglucan, are examined under the influence of dehydration and rehydration. Significant crystal structure changes were observed in the later stage of drying for both cotton and bacterial cellulose (BC). The 1 % lateral expansion in glucan chain spacing and 17 % decrease of calculated Scherrer dimension were detected for cotton due to the distortion of the structure possibly caused by mechanical stresses associated with drying. No detectable changes on average glucan chain spacings were observed for large BC crystals. However, an average width decrease by 4.4 nm was discovered in the (010) direction, which was more significant than that observed in the (100) and (110) directions. It is hypothesized that co-crystallized elementary fibrils preferentially disassociate along the (010) plane resulting in a significant reduction of crystal width. In the BC-xyloglucan model composite, the presence of xyloglucan does not interfere with the dehydration behavior. Rehydration leads to some structural changes but to a lesser extent than the initial drying. High temperature dehydration induced deformation and crystal size changes are found to be non-reversible due to the removal of the last hydration layer on the cellulose surface.     

Characterisation of bacterial cellulose from diverse <Emphasis Type="Italic">Komagataeibacter</Emphasis> strains and their application to construct plant cell wall analogues

This study investigated cellulose production and microstructure variation from six Komagataeibacter strains (ATCC 53524, ATCC 10245, ATCC 23769, ATCC 700178, NBRC 13693 and KTH 5655). Strain KTH 5655 produced the highest cellulose yields (10.39 g/l) after 9 days cultivation. Nuclear magnetic resonance spectroscopy and X-ray diffraction revealed that strain ATCC 23769 synthesised cellulose with the lowest crystallinity and decreased ratio of Iα/Iβ allomorph, whilst strain KTH 5655 produced a relatively ordered cellulose structure. However, the average widths of cellulose ribbons were similar (30–50 nm) for all types of cellulose. Phylogenetic analysis of the 16S rRNA gene indicated that these strains shared a high level of genetic similarity (ranging from 88 to 98%). All strains were used to produce cellulose in the presence of arabinoxylan or xyloglucan as simplified cell wall analogues. Our results provide guidance for the selection of cellulose-producing strains for specific biotechnological and research applications.     

Acetobacter cellulosic biofilms search for new modulators of cellulogenesis and native membrane treatments

Since natural substances like pseudoxanthins exert a positive effect on the cellulogenic ability ofAcetobacter xylinum when producing cellulosic pellicles suitable for skin burn therapy, new defined and complex modulators were sought. Ca²⁺ and Mg²⁺ (4 mM) were strongly stimulatory. Na⁺ had no effect and K⁺ was inhibitory. Ammonium dihydrogen phosphate (0.12 g/L) ensured the same nitrogen supply as the same concentration of yeast extract as measured by cellomembrane dry wt./yield albeit higher yeast extract supplies produced thicker membranes. Corn steep liquor (CSL) was also progressively beneficial from 0.125 to 0.5 mL/L, and this yield could be further improved by the combination of CSL with a tea infusion (source of caffeine). Uridine (precursor for UDP-Glc, sugar donor in cellulose biosynthesis), guanine, guanosine, and its butirylated derivatives (precursors for the positive modulator of cellulose synthetase, di-cGMP) resulted in only moderate stimulation. Sodium phytate and betaine were also slightly stimulatory. The fibrilar product from a newAcetobacter isolate (Ax-M) was characterized as cellulose by comparison with the solid-state¹³C-NMR of algal cellulose. Its X-ray diffractogram was a confirmatory analysis. After incorporation of tamarind xyloglucan to previously air-dried cellulosic pellicles, diffractometry displayed only slight differences. Mercerized (5M NaOH) fresh cellulosic biofilms underwent drastic size reduction (3.5-fold), turning compact nut still flexible if maintained wet.     

Dewetting pattern and stability of thin xyloglucan films adsorbed on silicon and mica

Thin polysaccharide films prepared with xyloglucan (XG), a neutral polysaccharide extracted from the seeds of Guibourtia hymenifolia were prepared by spin-coating and drop deposition under pH3, pH5 and pH12, on silicon and mica substrates. Atomic force microscopy (AFM) images show flat nanoporous matrices with additional grain-like structures on both mica and silicon for pH 3 and pH 5. However, X-ray photoelectron spectroscopy (XPS) and Auger spectra of these adsorbed biopolymers prepared under alkaline condition (pH 12) reveal that Na⁺ ions from the solution interact with the mica substrate surface and with XG forming chemical bonds. Both XPS and Auger results suggest XG depolymerisation during adsorption, caused by an alkaline ß-base catalyzed degradation mechanism, which is consistent with the more basic character of the mica surface under these conditions. Thus, the polysaccharide diffusion is inhibited during dewetting due to the surface bonding. On the other hand, the interaction of Na⁺ in solution with the silicon surface is weaker, favoring its interaction with the polysaccharide, conserving the overall polymer structure of XG and allowing the biopolymer to slip and diffuse during dewetting, forming the final branched fractal structure.     

Utilizing Polymer Blends to Prepare Ultrathin Films with Diverse Cellulose Textures

This paper presents an overview of the recent work on ultrathin polymer blend films containing cellulose. Three systems prepared via trimethylsilyl cellulose derivative, which is subsequently hydrolyzed to cellulose, are presented: polystyrene/cellulose, poly(methyl methacrylate)/cellulose and polystyrene-block-polyethyleneoxide/cellulose. Diverse textures emerge within the films depending on the interactions between the polymers and their interactions with the substrate as well as on different solubilities of the polymers. Furthermore, an ultrathin film containing a cellulose/xylan blend is presented. This film was deposited directly from a common solvent (dimethylacetamide/lithium chloride) and it did not exhibit distinct morphological patterns comparable to the blends with synthetic polymers.