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Polysaccharides have attracted immense attention as the largest source of bioactive compounds. Its bioavailability and bioactivity can be improved by utilizing degradation enzymes to reduce their molecular weight and viscosity. In this study, a 654 bp gene encoding xylanase was screened from the genome of Bacillus altitudinis JYY-02 and overexpressed in Escherichia coli Rosetta (DE3). The recombinant xylanase with a molecular weight of 27.98 kDa was purified (11.7-fold) using Ni-NTA affinity chromatography, with a 43.6% final yield. Through molecular docking, Glu, Arg, Tyr, and Trp were found to be the main amino acids involved in the interaction between xylanase and xylobiose. The effects of pH, temperature, metal ions, and substrates on xylanase activity were determined, and the results showed that the highest catalytic activity was displayed at pH 6.5, 50 °C temperature, with Cu 2+ as an activator and xylan as the substrate. The K m (substrate concentration that yields a half-maximal velocity) and V max (maximum velocity) of recombinant xylanase were 6.876 mg/mL and 10984.183 μmol/mg∙pr/min, respectively. The recombinant xylanase was thermostable, with 85% and 39% of the enzymatic activity retained after 1 h at 60 °C and 1 h at 90 °C, respectively. The recombinant xylanase demonstrated a significant clarifying effect on fruit juices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Renewable materials of biological origin exhibit attractive properties in relation to traditional plastics, as they can be partially or completely replaced, thereby reducing environmental impacts. Hemicelluloses are a group of polysaccharides that have expanded applications when acetylated. Acetylation can improve the mechanical strength and water vapor barrier properties of xylan-based bioplastics. By partially acetylating xylan in the present study, it was possible to use water as a solvent for the film-forming solution and starch as a second polysaccharide in the formation of bioplastics. Xylan was modified via partial chemical acetylation by varying the reaction time, solvent, and catalyst content. The bioplastics were formed by non-acetylated xylan and acetylated xylan with degrees of substitution (DS) of 0.45 and 0.9, respectively, with starch to form blends using glycerol as a plasticizer. Acetylation with DS 0.45 showed better results in increasing the hydrophilicity of the bioplastic. On the other hand, acetylation influenced the thermal stability of bioplastics, increasing the maximum temperature of the degradation rate from 302 °C to 329 °C and 315 °C, owing to changes in the crystallinity of the polymers. In addition to the modulus of elasticity 2.99 to 290.61 and 274.67 MPa for the non-acetylated bioplastic and the bioplastic with DS of 0.45 and 0.90, respectively. Thus, the films obtained presented suitable physicochemical properties for use in various industrial applications, such as active and intelligent packaging in the food sector., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Structural elucidation of plant cell wall xyloglucan through the analysis of enzymatically produced fragments requires detailed knowledge of enzymes hydrolytic mechanism. In this note, the mode of action and cleavage site of commercial recombinant xyloglucanases (GH74, Paenibacillus sp.) was studied on native and fluorescent-tagged tamarind xyloglucan. In complement to information provided by the manufacturer, GH74 hydrolysis was shown dual endo/exo- and exo-processive with low affinity towards labelled reducing-ends. GH74 accommodated X/G in its subsite -1 and X/L in its subsite +1. Moreover, hydrolysis kinetic indicated a GH74 activity inhibition by excess products. These results will help for application of this enzyme in xyloglucans structural analysis or for processing cell walls., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this study, novel redox-sensitive nanoparticles based on xylan-lipoic acid (Xyl-LA) conjugate were developed for tumor targeted delivery of niclosamide (Nic) in cancer therapy. The niclosamide loaded xylan-lipoic acid conjugate nanoparticles (Xyl-LA/Nic NPs) showed redox responsive behaviour in presence of reductive glutathione (GSH), which indicate their suitability for intracellular drug release. The obtained Xyl-LA/Nic NPs exhibited uniform particle size (196 ± 1.64 nm), high loading capacity (~28.6 wt %) and excellent blood compatibility. The anticancer activity of the Niclosamide and the Xyl-LA/Nic NPs against the colon carcinoma cell lines (HCT-15, Colo-320) were evaluated by MTT assay and the overall results indicate that the Xyl-LA/Nic NPs significantly enhanced the therapeutic efficiency of niclosamide in cancer therapy., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hemicellulose-rich substrates produced in the lignocellulose biorefinery context can yield macromolecular xylan structures with assorted application in the chemical industry. Xylan presents natural affinity to cellulose and its incorporation onto fibers increases the physical processability of pulp; however, current studies diverge on how molar mass affects xylan interaction with cellulose. In the current work, xylans with varied structural characteristics were prepared from alkaline-sulfite pretreated sugarcane bagasse with aid of an alkaline-active xylanase and selective precipitations using different ethanol concentrations. Prepared xylan fractions, containing low levels of lignin contamination (4-9%) and molar masses ranging from 2.3 kDa to 34 kDa, were incorporated onto eucalyptus pulp fibers up to 4.7 g xylan/100 g pulp. The efficiency of xylan incorporation onto cellulosic fibers was dependent on the xylan structures, where low molar mass and low substitution degree favored high incorporation levels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Guavira is a plant that belongs to Myrtaceae family, being widespread in the Brazilian Cerrado. In this study, pectic and hemicellulosic polysaccharides from guavira pomace, an agroindustry residue from pulp production, were structurally characterized using GPC, monosaccharide composition, methylation and NMR experiments. The absolute configurations of monosaccharides and the nature of uronic acids were attributed according to numerous data on the composition of related plant arabinogalactans and hemicelluloses present in the literature. An arabinan was purified, presenting Ara (85.0%), Rha (3.3%), Gal (7.7%) and GalA (4.0%). Mono and bidimensional NMR analyses of this arabinan demonstrated the presence of terminal β-L-Araf units, whose occurrence has been scarcely reported in the literature. Hemicellulosic fraction contained a glucuronoxylan, with α-D-GlcpA/4-O-methyl-α-D-GlcpA group linked to O-2 of a (1 → 4)-β-D-xylan, presenting one uronic acid residue for every six xylose units. These findings about guavira pomace polysaccharides could contribute to develop future nutraceutical and technological uses for this industrial waste., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Symbionts in the gut of termites are expected to be large sources of enzymes involved in lignocellulose degradation, but their biotechnological potential has not been fully explored. In this study, we expressed, purified, and biochemically characterized a glycoside hydrolase family 11 xylanase, NtSymX11, from a symbiotic bacterium of the higher termite, Nasutitermes takasagoensis. NtSymX11 is a multimodular enzyme consisting of a catalytic domain and two tandem carbohydrate-binding modules (CBM36). The pH and temperature optima of NtSymX11 were pH 6.0 and 40 °C, respectively. By comparing the properties of full-length and truncated variants of NtSymX11, it was shown that CBM36 decreases the enzyme stability at acidic pH and high temperature. The main products from xylohexaose and various xylan substrates were X1-X3 xylooligosaccharides. Analysis of kinetic parameters indicated that NtSymX11 displays an outstanding catalytic performance when compared to other reported xylanases, and CBM36 enhances the activity by increasing the affinity to the substrate. Addition of Ca 2+ boosted the activity of full-length enzyme, but not the truncated variant lacking the CBM, against the insoluble substrate, suggesting that CBM36 plays a role in the Ca 2+ -dependent increase of catalytic efficiency., (Copyright © 2019. Published by Elsevier Ltd.)

Several tetraalkylphosphonium and tetraalkylammonium salts containing xyloside- and xylobioside-based anionic moieties have been prepared. Two stereoselective routes have been developed: i) a chemical pathway in four steps from D-xylose, and ii) a chemoenzymatic pathway directly from biomass-derived xylans. These salts displayed interesting properties as ionic liquids. Their structures have been correlated to their thermal properties (melting, glass transition and decomposition temperatures)., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Arabinoxylans (AX) are a major class of hemicellulose and an important polysaccharide component of lignocellulosic biomass. To utilize the glycan polymer effectively, it is desirable to learn more about the enzymatic hydrolysis of AXs. Well-defined glycans can help to elucidate these processes. Here, we report the efficient synthesis of a mixed O- and S-linked tetraxylan. This thio-oligosaccharide has been developed as a putative inhibitor of arabinoxylan degrading enzymes used for the saccharification of biomass. Two common approaches for the synthesis of thio-oligosaccharides, either involving 1-thioglycoside donors or thioacceptors, are presented and compared regarding byproduct formation and yields. Both methods have shown to be useful for the synthesis of thiolinkages in oligoxylans assembly. However, the success of the reaction is highly dependent on the "match" between donors and acceptors., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A statistical model describing the random distribution of substituted xylopyranosyl residues in arabinoxylooligosaccharides is suggested and compared with existing experimental data. Structural diversity of arabinoxylooligosaccharides of various length, originating from different arabinoxylans (wheat flour arabinoxylan (arabinose/xylose, A/X = 0.47); grass arabinoxylan (A/X = 0.24); wheat straw arabinoxylan (A/X = 0.15); and hydrothermally pretreated wheat straw arabinoxylan (A/X = 0.05)), is semiquantitatively approximated using the proposed model. The suggested approach can be applied not only for prediction and quantification of arabinoxylooligosaccharides' structural diversity, but also for estimate of yield and selection of the optimal source of arabinoxylan for production of arabinoxylooligosaccharides with desired structural features., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A range of α-L-arabinofuranosyl-(1-4)-β-D-xylo-oligosaccharides (AXOS) were produced by hydrolysis of wheat flour arabinoxylan (WAX) and acid debranched arabinoxylan (ADWAX), in the presence and absence of an AXH-d3 α-L-arabinofuranosidase, by several GH10 and GH11 β-xylanases. The structures of the oligosaccharides were characterised by GC-MS and NMR and by hydrolysis by a range of α-L-arabinofuranosidases and β-xylosidase. The AXOS were purified and used to characterise the action patterns of the specific α-L-arabinofuranosidases. These enzymes, in combination with either Cellvibrio mixtus or Neocallimastix patriciarum β-xylanase, were used to produce elevated levels of specific AXOS on hydrolysis of WAX, such as 3(2)-α-L-Araf-(1-4)-β-D-xylobiose (A(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(3)XX), 2(2)-α-L-Araf-(1-4)-β-D-xylotriose (XA(2)X), 3(2)-α-L-Araf (1-4)-β-D-xylotriose (XA(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotriose (A(2+3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2+3)XX), 2(4),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(2+3)XXX) and 3(3),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(3)A(3)XX), many of which have not previously been produced in sufficient quantities to allow their use as substrates in further enzymic studies. For A(2,3)XX, yields of approximately 16% of the starting material (wheat arabinoxylan) have been achieved. Mixtures of the α-L-arabinofuranosidases, with specific action on AXOS, have been combined with β-xylosidase and β-xylanase to obtain an optimal mixture for hydrolysis of arabinoxylan to L-arabinose and D-xylose., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Switchgrass (Panicum virgatum, L.) is a potential renewable source of carbohydrates for use in microbial conversion to biofuels. Xylan comprises approximately 30% of the switchgrass cell wall. To understand the limitations of commercial enzyme mixtures, alkali-extracted, isolated switchgrass xylan was hydrolyzed by the action of two commercial enzyme cocktails, in the presence and absence of an additional α-arabinofuranosidase enzyme. The two most abundant enzymatic digestion products from each commercial enzyme treatment were separated and characterized by LC-MS(n), linkage analysis, and NMR. The most abundant oligosaccharide from each commercial cocktail was susceptible to hydrolysis when supplemented with a GH62 α-arabinofuranosidase enzyme; further characterization confirmed the presence of (1→3)-α-arabinose linkages. These results demonstrate the lack of the required selectivity for arabinose-containing substrates in the commercial enzyme preparations tested. One product from each condition remained intact and was found to contain (1→2)-β-xylose-(1→3)-α-arabinose side chains; this linkage acts as a source of oligosaccharide recalcitrance., (Published by Elsevier Ltd.)

In comparison to the annual grain crops dominating current agricultural production, perennial grain species require fewer chemical and energy inputs and improve soil health and erosion control. The possibility for producing sustainable grain harvests from marginal land areas is motivating research initiatives to integrate perennial grains into commercial cropping and food processing systems. In this study, the feruloylated arabinoxylans from intermediate wheat grass (Thinopyrum intermedium, IWG), a promising perennial grain candidate in agronomic screening studies, were investigated. Insoluble fiber isolated from IWG whole grain flour was subjected to either mildly acidic (50 mM TFA, 100 °C, 2 h) or enzymatic (Driselase) hydrolysis. The liberated feruloylated arabinoxylan oligosaccharides were concentrated with Amberlite XAD-2, separated with gel chromatography (Sephadex LH-20, water), and purified with reversed-phase HPLC (C18, water-MeOH gradient). Thirteen feruloylated oligosaccharides were isolated (including eight structures described for the first time) and identified by LC-ESI-MS and NMR. Linkage-type analysis via methylation analysis, as well as the monosaccharide and phenolic acid profiles of the IWG insoluble fiber were also determined. IWG feruloylated arabinoxylans have a relatively simple structure with only short feruloylated side chains, a lower backbone substitution rate than annual rye and wheat varieties, and a moderate phenolic acid content., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Birch xylan was functionalized with various thiols through UV initiated radical thiol-ene reaction under mild conditions. Xylan was allylated through etherification with allyl glycidyl ether under alkaline conditions. The allylated xylan was then reacted with thiols containing varying functional groups: trimethylbenzyl mercaptan, dodecanethiol, thioglycolic acid, L-cysteine and cysteamine hydrochloride. The reactions were conducted under homogeneous conditions at room temperature, either in water (hydrophilic thiols) or in DMF (hydrophobic thiols). The effect of reaction parameters to the functionalization efficiency was studied, including, for example, thiol excess, thiol character, initiator amount and reaction mixture concentration. The reactions were fast and 100% conversion of allyl groups was reached in most cases, sometimes already within 10 min. Water as solvent resulted generally in faster reactions when compared to DMF, and it was possible to conduct the aqueous reaction even without added UV initiator. It was also possible to incorporate two functionalities simultaneously during one reaction into the xylan structure., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 1H chemical shifts were assigned using two-dimensional correlation spectroscopy. A comprehensive update of the nomenclature describing xyloglucan side-chain structures is provided for reference., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Switchgrass (Panicum virgatum, L.) is a potential dedicated biomass crop for use in biocatalytic conversion systems to biofuels. Nearly 30% of switchgrass cell wall material is xylan. The complete depolymerization of xylan is desirable both as an additional carbon source for microbial fermentation and to reduce inhibitory effects xylooligomers may have on cellulolytic glycoside hydrolase enzymes. To identify structural features of switchgrass xylan that are not distinguishable by mass spectrometry alone, a α-arabinofuranosidase enzyme was used to remove the arabinose side chains from alkali-extracted switchgrass xylan from three cultivars with simultaneous hydrolysis by β-endo-xylanase to enrich for oligosaccharide products with extended branching. The two most abundant enzymatic digestion products were separated and characterized by LC-MS(n), linkage analysis, and NMR. These two oligosaccharides were present in all three switchgrass cultivars and found to contain (1→2)-β-xylose-(1→3)-α-arabinose side chains, a linkage not previously reported in switchgrass., (Published by Elsevier Ltd.)

COB1B1S2 was isolated from an alkaline extract of Cassia obtusifolia seeds, and purified by anion-exchange and gel permeation chromatography. It contains arabinose, xylose, and glucuronic acid, in the molar ratio of 5:81:14, with an apparent molecular weight estimated to be 70.4 kDa. Elucidated by using chemical and spectroscopic methods, COB1B1S2 was shown to have a backbone consisting of 1,4-linked β-D-Xylp, with one single-unit terminal α-D-GlcpA or α-L-Araf substituted at O-2 for nearly every five 1,4-linked Xylp. COB1B1S2 is structurally different from typical glucuronoxylans by its absence of methylation at O-4 of GlcA. The native COB1B1S2 showed no significant inhibition on the tube formation of human microvascular endothelial cells (HMEC) and on the growth of liver and colon cancer cells. On the contrary, COB1B1S2-Sul, prepared as the sulfated derivative of COB1B1S2, exhibited a significant inhibition on tube formation of HMEC in a dose-dependent manner, and on the growth of Bel7402 liver cancer cells. These results indicated that the introduction of sulfate groups significantly enhanced the biological activity of glucuronoxylan., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Walls from wheat (Triticum aestivum L.) endosperm are composed primarily of hetero-(arabino)xylans (AXs) (70%) and (1→3)(1→4)-β-D-glucans (20%) with minor amounts of cellulose and heteromannans (2% each). To understand the differential solubility properties of the AXs, as well as aspects of their biosynthesis, we are sequencing the xylan backbone and examining the reducing end (RE) sequence(s) of wheat (monocot) AXs. A previous study of grass AXs (switchgrass, rice, Brachypodium, Miscanthus and foxtail millet) concluded that grasses lacked the comparable RE glycosyl sequence (4-β-D-Xylp-(1→4)-β-D-Xylp-(1→3)-α-L-Rhap-(1→2)-α-D-GalpA-(1→4)-D-Xylp) found in dicots and gymnosperms but the actual RE sequence was not determined. Here we report the isolation and structural characterisation of the RE oligosaccharide sequence(s) of wheat endosperm cell wall AXs. Walls were isolated as an alcohol-insoluble residue (AIR) and sequentially extracted with hot water (W-sol Fr) and 1M KOH containing 1% NaBH4 (KOH-sol Fr). Detailed structural analysis of the RE oligosaccharides was performed using a combination of methylation analysis, MALDI-TOF-MS, ESI-QTOF-MS, ESI-MS(n) and enzymic analysis. Analysis of RE oligosaccharides, both 2AB labelled (from W-sol Fr) and glycosyl-alditol (from KOH-sol Fr), revealed that the RE glycosyl sequence of wheat endosperm AX comprises a linear (1→4)-β-D-Xylp backbone which may be mono-substituted with either an α-L-Araf residue at the reducing end β-D-Xylp residue and/or penultimate RE β-D-Xyl residue; β-D-Xylp-(1→4)-[α-L-Araf-(1→3)](+/-)-β-D-Xylp-(1→4)-[α-L-Araf-(1→3)](+/-)-β-D-Xylp and/or an α-D-GlcpA residue at the reducing end β-D-Xylp residue; β-D-Xylp-(1→4)-[α-L-Araf-(1→3)](+/-)-β-D-Xylp-(1→4)-[α-D-GlcAp-(1→2)]-β-D-Xylp. Thus, wheat endosperm AX backbones lacks the RE sequence found in dicot and gymnosperm xylans; a finding consistent with previous reports from other grass species., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Cellulose is an attractive supporting matrix for diverse biotechnological applications, including chromatography, diagnostics, and tissue replacement/scaffolding, due to its renewable resource status, low cost, and low non-specific interaction with biomolecules. In an effort to expand the biofunctionality of cellulose materials, we present here a versatile method for the synthesis of xyloglucan-peptide conjugates that harness the strong xyloglucan-cellulose binding interaction for gentle surface modification. Xylogluco-oligosaccharide aminoalditols (XGO-NH(2)) were coupled to both linear and cyclic peptides, which contained the endothelial cell epitope Arg-Gly-Asp, in a facile two-step approach employing diethyl squarate cross-linking. Subsequent xyloglucan endo-transglycosylase-mediated coupling of the resulting XGO-GRGDS (Gly-Arg-Gly-Asp-Ser) and XGO-c[RGDfK]-PEG-PEG (cyclo[Arg-Gly-Asp-(D-Phe)-Lys]-PEG-PEG; where PEG is 8-amino-3,6-dioxaoctanoic acid) conjugates into high molecular mass xyloglucan yielded xyloglucan-RGD peptide conjugates suitable for cellulose surface activation. Notably, use of XGO-squaramate as a readily accessible, versatile intermediate overcomes previous limitations of solid-phase synthetic approaches to XGO-peptide conjugates, and furthermore allows the method to be generalized to a wide variety of polypeptides and proteins, as well as diverse primary amino compounds., (Copyright © 2012. Published by Elsevier Ltd.)

The esterification of xylan type hemicelluloses, isolated from birchwood, was carried out firstly in homogeneous conditions using N,N-dimethylformamide (DMF) and lithium chloride (LiCl) in the presence of 4-dimethylaminipyridine (DMAP). The degree of substitution (DS) of xylan acetates ranged between 0.9 and 2.0 as a function of experimental conditions. Due to the problems of toxicity and recycling of DMF, an alternative method of esterification is reported in the second part of this work, performing in the absence of organic solvent and using DMAP or methanesulfonic acid (MSA) as catalysts. Acetylation reaction catalyzed by MSA was developed through an experimental design in order to achieve the highest DS under the mildest conditions. The significant factors and their interactions were identified. The optimization of reaction parameters allowed to obtain a high DS (1.6) and maximal yield (85%). Moreover, the reactivity of propionic and hexanoic anhydrides was evaluated and hydrophobic xylan esters with low degrees of substitution were obtained., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The interaction between para-crystalline cellulose and the cross-linking glycan xyloglucan (XG) plays a central role for the strength and extensibility of plant cell walls. The coating of XGs on cellulose surfaces is believed to be one of the most probable interaction patterns. In this work, the effects of explicit water and side chain variation on the adsorption of XGs on cellulose are investigated by means of atomistic molecular dynamics simulations. The adsorption properties are studied in detail for three XGs on cellulose Iβ 1-10 surface in aqueous environment, namely GXXXGXXXG, GXXLGXXXG, and GXXFGXXXG, which differ in the length and composition of one side chain. Our work shows that when water molecules are included in the theoretical model, the total interaction energies between the adsorbed XGs and cellulose are considerably smaller than in vacuo. Furthermore, in water environment the van der Waals interactions prevail over the electrostatic interactions in the adsorption. Variation in one side chain does not have significant influence on the interaction energy and the binding affinity, but does affect the equilibrium structural properties of the adsorbed XGs to facilitate the interaction between both the backbone and the side chain residues with the cellulose surface. Together, this analysis provides new insights into the nature of the XG-cellulose interaction, which helps to further refine current molecular models of the composite plant cell wall., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Xylans from five seaweeds belonging to the order Nemaliales (Galaxaura marginata, Galaxaura obtusata, Tricleocarpacylindrica, Tricleocarpa fragilis, and Scinaia halliae) and one of the order Palmariales (Palmaria palmata) collected on the Brazilian coasts were extracted with hot water and purified from acid xylomannans and/or xylogalactans through Cetavlon precipitation of the acid polysaccharides. The β-D-(1→4), β-D-(1→3) 'mixed linkage' structures were determined using methylation analysis and 1D and 2D NMR spectroscopy. The presence of large sequences of β-(1→4)-linked units suggests transient aggregates of ribbon- or helical-ordered structures that would explain the low optical rotations., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Fluorescently labeled derivatives of a xyloglucan (XG) nonasaccharide Glc(4)Xyl(3)Gal(2) (XLLG) were used as glycosyl acceptors in assays of xyloglucan endotransglycosylase (XET) from germinated nasturtium (Tropaeolum majus) seeds. We have investigated how the type of the oligosaccharide label influences the kinetic parameters of the reaction. The fluorescent probes used to label XLLG were anthranilic acid (AA), 8-aminonaphtalene-1,3,6-trisulfonic acid (ANTS), fluorescein isothiocyanate (FITC), and sulforhodamine (SR), respectively. The obtained data were compared with those of the reactions where aldose and/or alditol forms of tritium-labeled xyloglucan-derived nonasaccharide served as the respective acceptors. Modification at C-1 of the reducing-end glucose in XLLG by substitution with the fluorophore markedly affected the kinetic parameters of the reaction. The Michaelis constants K(m) for individual acceptors increased in the order [1-(3)H]XLLGXLLG-SR>XLLG-ANTS>[1-(3)H]XLLGol>[1-(3)H]XLLG>XLLG-AA. Catalytic efficiency (expressed as k(cat)/K(m)) with XLLG labeled with SR or FITC was 15 and 28 times, respectively, higher than with the tritium-labeled natural substrate [1-(3)H]XLLG. Comparison of the kinetic parameters found with acceptors labeled with different types of labels enables to select the most effective substrates for the high-throughput assays of XET., (2010 Elsevier Ltd. All rights reserved.)

Xylan has been used as a raw material in the synthesis of butyl, octyl and decyl glycosides. Mixtures of D-xylose-, L-arabinose- and D-glucose-based surfactants were obtained under smooth conditions with high yields in a one-pot process. The surface activities of octyl and decyl glycosides thus obtained have been studied and compared with that of pure alkyl D-xylosides. The results have confirmed that the new synthetic approach described in this paper is a potentially economical and efficient method for the preparation of environmentally friendly surfactants., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Ball-milled alcohol-insoluble residue (AIR) was prepared from switchgrass (Panicum virgatum var Alamo) and sequentially extracted with 50 mM ammonium oxalate buffer, 50 mM sodium carbonate, 1 M KOH containing 1% NaBH(4), and 4 M KOH containing 1% NaBH(4). Arabinoxylan was the most abundant component of the 1 M KOH-extracted fraction, which was treated with endoxylanase to generate oligosaccharides. Gel-permeation chromatography of these oligosaccharides produced three size-homogeneous oligosaccharide fractions with molecular weights of 678, 810, and 1074 Da, corresponding to 5, 6, and 8 pentose units, respectively. Detailed structural analysis of these oligosaccharides was performed using methylation analysis, multiple-step mass spectrometry (ESIMS(n)), and 1D and 2D NMR spectroscopy. The preferred gas-phase fragmentation pathways were identified for these oligosaccharides, providing extensive sequence information that was completely consistent with structures determined by ab initio NMR analysis. These results demonstrate the high information content of ESIMS(n) analysis when applied to cell-wall-derived oligosaccharides and provide standard data that will facilitate the analysis of cell-wall polysaccharide fragments with a sensitivity that is sufficient for the analysis of samples obtained from dissected tissues as well as other small samples., (Copyright © 2010. Published by Elsevier Ltd.)

Destarched and deproteinated water-unextractable material (WUM) of rye outer layers was sequentially treated with lichenase and cellulase to digest beta-glucans and a part of the cellulose. As a result, the polymeric cell-wall material (CWM) initially associated with these polysaccharides was released into solution (AXL and AXC for lichenase- and cellulase-extractable fractions, respectively). A portion of the material that self-aggregated during extractions was further solubilized with DMSO (XD and XD-P for the fraction left in the solution and that precipitated during dialysis, respectively). Arabinoxylans (AXs) recovered from these fractions were composed of populations with different degrees of substitution with alpha-l-arabinofuranosyl residues (Araf). Their counterparts present in the AXL and AXC fractions exhibited higher (0.60 and 0.75) arabinose-to-xylose ratios (Ara/Xyl) and represented 27% and 32% of the isolated AXs, respectively. The xylans of the XD and XD-P fractions had a very low Ara/Xyl ratio (0.16 and 0.09) and accounted for 23% and 18%, respectively. Based on the results of ammonium sulfate fractionation and sugar analysis, it has been shown that AXL consisted of AX subfractions having Ara/Xyl in a narrow range (0.50-0.66). By contrast, the cellulase-extractable AXs were characterized by the presence of the highly branched subfractions (Ara/Xyl of 1.00) as well. Quite unexpectedly, the higher amounts of ferulic acid (FA) were found in the cell-wall fractions enriched in xylans than in the AX-containing fractions. Furthermore, as demonstrated by (1)H NMR and Fourier transform infrared spectroscopy, xylans were substituted with alpha-d-glucuronopyranosyl residues (GlcpA).

The presence of a fairly uncommon side chain 2-O-beta-D-xylopyranosyl-alpha-L-arabinofuranosyl in arabinoxylans (AX) from eight different cereal by-products was investigated, using (1)H NMR spectroscopy and high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) after Shearzyme (GH10 endo-1,4-beta-D-xylanase) hydrolysis. This disaccharide side group was present in significant amounts in AX extracted from corn cobs and barley husks. For the first time, it was also detected in AX from oat spelts and rice husks, and in lesser amounts in wheat straw AX. Arabinoxylo-oligosaccharide (AXOS) containing the 2-O-beta-D-Xylp-alpha-L-Araf side chain was purified from the oat spelt AX hydrolysate and the structure was fully analyzed using 1D and 2D NMR spectroscopy. The AXOS was identified as beta-D-Xylp-(1-->2)-alpha-L-Araf-(1-->3)-beta-D-Xylp-(1-->4)-D-Xyl. To our knowledge, such a structure with 2-O-beta-D-Xylp-alpha-L-Araf attached to the O-3 of the nonreducing end of xylobiose has not been described previously. New information on substitution of AX from various cereal by-products was obtained by combining NMR and enzyme-assisted HPAEC-PAD analysis.

Biochemical studies to elucidate the structural basis for xyloglucan specificity among GH12 xyloglucanases are lacking. Accordingly, the substrate specificity of a GH12 xyloglucanase from Aspergillus niger (AnXEG12A) was investigated using pea xyloglucan and 12 xylogluco-oligosaccharides, and data were compared to a structural model of the enzyme. The specific activity of AnXEG12A with pea xyloglucan was 113 micromol min(-1)mg(-1), and apparent k(cat) and K(m) values were 49 s(-1) and 0.54 mg mL(-1), respectively. These values are similar to previously published results using xyloglucan from tamarind seed, and suggest that substrate fucosylation does not affect the specific activity of this enzyme. AnXEG12A preferred xylogluco-oligosaccharides containing more than six glucose units, and with xylose substitution at the -3 and +1 subsites. The specific activities of AnXEG12A on 100 microM XXXGXXXG and 100 microM XLLGXLLG were 60+/-4 and 72+/-9 micromol min(-1)mg(-1), respectively. AnXEG12A did not hydrolyze XXXXXXXG, consistent with other data that demonstrate the requirement for an unbranched glucose residue for hydrolysis by this enzyme.

Different agricultural wastes, namely tobacco stalk (TS), cotton stalk (CS), sunflower stalk (SS), and wheat straw (WS), were used for the production of xylooligosaccharide (XO). XO production was performed by acid hydrolysis of xylan, which was obtained by alkali extraction from these agricultural wastes. The major component of these agricultural wastes was determined as cellulose (30-42%), followed by xylan (20%) and lignin (20-27%). Xylans from these wastes had mainly xylose (85-96%) with small amount of glucose, while wheat straw xylan contained also arabinose. The best xylan conversion into XOs was achieved with 0.25M H(2)SO(4) with 30-min reaction time. Under these conditions, the XO yield was between 8% and 13%. The yield of XOs depends on both acid concentration and hydrolysis time, but the yield of monosaccharide depends on the structure and composition of xylan besides acid concentration and the time. The more branched xylan, WSX, gave the highest monosaccharide ( approximately 16%) and furfural ( approximately 49mg/100g xylan) yield. This research showed that all xylans from selected agricultural wastes generated XOs with similar profiles, and these oligosaccharides could be used as functional food ingredients or soluble substrates for xylanases.

The current study forms part of an ongoing research effort focusing on the elucidation of the chemical structure of the sulfated extracellular polysaccharide of the red microalga Porphyridium sp. (UTEX 637). We report here on the chemical structure of a fraction separated from an acidic crude extract of the polysaccharide, as investigated by methylation analysis, carboxyl reduction-methylation analysis, desulfation-methylation analysis, partial acid hydrolysis, Smith degradation, together with 1D and 2D (1)H and (13)C NMR spectroscopy. This fraction with a molar mass of 2.39x10(5)g mol(-1) comprised D- and L-Gal, D-Glc, D-Xyl, D-GlcA, and sulfate groups in a molar ratio of 1.0:1.1:2.1:0.2:0.7. The almost linear backbone of the fraction is composed of (1-->2)- or (1-->4)-linked d-xylopyranosyl, (1-->3)-linked L-galactopyranosyl, (1-->3)-linked D-glucopyranosyl, and (1-->3)-linked D-glucopyranosyluronic acid and comprises a possible acidic building unit: [(2 or 4)-beta-D-Xylp-(L-->3)]m-alpha-D-Glcp-(1-->3)-alpha-D-GLCPA-(1-->3)-L-Galp(l-->. Attached to the backbone are sulfate groups and nonreducing terminal D-xylopyranosyl and galactopyranosyl residues, which occur at the O-6 positions of Glc-derived moieties in the main chain.

Three d-xylan type per-O-methylated trisaccharides with various types of linkages between the d-xylopyranose units were examined by atmospheric pressure photoionization (APPI) mass spectrometry in the positive ion mode. The most interesting feature of a thermospray mass spectrum using the APPI source with UV lamp switched off, is the exclusive production of [M+Na](+) adduct ions. [M+Na](+) cationized ions are the most abundant species in the case of APPI mass spectrometry. The second ionization process has no analogy in the case of substances studied using APPI to date. This aspect involves the addition of a water molecule to the molecular ion of a per-O-methylated saccharide, giving rise to [M+H(2)O](+) adduct ions. The [M+H(2)O](+) species are readily detected at m/z 544, and are clearly visible for all three isomers studied. The MS/MS spectrum of [M+Na](+) ions contains a base peak at m/z 375, produced by a Y-type cleavage of the trisaccharide, along with a hydrogen rearrangement on the terminal interglycosidically linkage glycosidic oxygen atom. The [M+H(2)O](+) species fragment largely give rise to ions at m/z 175, 143 and, as a result, the m/z 111 ion is unique to nonreducing terminal units.

This study was undertaken to investigate the effect of arabinose content on film properties. The substrate used was a rye arabinoxylan that had an Ara/Xyl ratio of 0.52 and an average number molecular weight of 305 kDa. Oxalic acid was used to attempt selective removal of the arabinose substituents on the xylan main chain. The debranching of the polymer was coupled with a decrease in molecular weight. The effect of reaction conditions on the decrease in arabinose content and loss of molecular weight was investigated. Optimal conditions were selected using an experimental design. Treatment at lower temperature for longer period of times resulted in debranching with less degradation of the main chain. As the Ara/Xyl ratio was lowered, aggregates began to form in an interval of the Ara/Xyl ratio between 0.31 and 0.23 in a water solution. Precipitation occurred below an Ara/Xyl ratio of 0.1. Thus, removal of arabinose substituents results in a gradual association of unsubstituted chains. There is a linear correlation between arabinose substitution and the moisture content of arabinoxylan at 98% RH. A decrease in arabinose content resulted in the loss of a plasticizing effect, as determined by dynamic mechanical analysis, which is correlated to water binding capacity.

Arabinoxylans (AX) are the predominant cell-wall polysaccharides in wheat flour. Water-extractable AX are essential for dough and bread properties and performance. However, there is no specific and accurate way of determining the content and structure of AX. An enzyme-assisted method employing an efficient enzyme mixture for the total hydrolysis of AX was developed in the present work. Enzymatic hydrolysis (EH) is a gentle method during which no unwanted sugar destruction occurs. Following EH, liberated monosaccharides were analysed by gas chromatography (GC) and liquid chromatography using HPAEC-PAD. The results were compared with acid methanolysis (AM) and acid hydrolysis (AH). EH performed better on commercially isolated AX samples than the reference method AM. Its action in the water extract from wheat flour was also more efficient than that of AM and comparable to the efficiency of AH. HPAEC-PAD revealed a significant amount of fructose in the water extract following EH, originating from fructans in wheat flour not detected in the GC analysis. The wheat flour examined contained 0.29% water-extractable AX. The arabinose/xylose ratio was 0.32. The enzyme-based method developed is applicable for comparison of different wheat flours and can be used in evaluating the effect of processing on the content and structure of water-extractable AX.

- Model composites, produced using cellulose from stationary cultures of the bacterium Gluconoacetobacter xylinus and tamarind xyloglucan, were examined by wide-angle X-ray scattering (WAXS) and CP/MAS solid-state (13)C NMR spectroscopy. The dominant crystallite allomorph of cellulose produced in culture media with or without xyloglucan was cellulose I(alpha) (triclinic). The presence of xyloglucan in the culture medium reduced the cross-section dimensions of the cellulose crystallites, but did not affect the crystallite allomorph. However, when the composites were refluxed in buffer, the proportion of cellulose I(beta) allomorph increased relative to that of cellulose I(alpha). In contrast, cellulose I(alpha) remained the dominant form when cellulose, produced in the absence of xyloglucan, was then heated in the buffer. Hence the presence of xyloglucan has a profound effect on the formation of the cellulose crystallites by G. xylinus.

The heteroxylan from the hybrid Paulownia elongata/Paulownia fortunei is an O-acetyl-(4-O-methylglucurono)xylan with an acetylation degree (DS) of 0.59 and a molecular weight (M(w)) of 29 kDa. The heteroxylan backbone is composed by (1-->4)-linked beta-d-xylopyranosyl units (Xylp) partially ramified with terminal (1-->2)-linked 4-O-methyl-alpha-D-glucuronosyl (MeGlcpA) and a small proportion of alpha-D-glucuronosyl (GlcpA) residues in a molar ratio of Xylp:(MeGlcpA+GlcpA) of 20:1. Roughly half of the beta-D-xylopyranosyl units in the backbone are acetylated: 3-O-acetylated (22 mol %), 2-O-acetylated (23 mol %) or 2,3-di-O-acetylated (7 mol %). ESI-MS and MALDI-MS studies of partially hydrolyzed heteroxylan revealed a random distribution of O-Ac and MeGlcpA within the backbone. However, the frequency of substitution with O-Ac along the backbone is not uniform and the molecular regions that did not contain MeGlcpA substituents possessed an acetylation degree significantly lower than the average DS of the xylan.

Isolated cell walls of Argania spinosa fruit pulp were fractionated into their polysaccharide constituents and the resulting fractions were analysed for monosaccharide composition and chemical structure. The data reveal the presence of homogalacturonan, rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II) in the pectic fraction. RG-I is abundant and contains high amounts of Ara and Gal, indicative of an important branching in this polysaccharide. RG-II is less abundant than RG-I and exists as a dimer. Structural characterisation of xyloglucan using enzymatic hydrolysis, gas chromatography, MALDI-TOF-MS and methylation analysis shows that XXGG, XXXG, XXLG and XLLG are the major subunit oligosaccharides in the ratio of 0.6:1:1.2:1.6. This finding demonstrates that the major neutral hemicellulosic polysaccharide is a galacto-xyloglucan. In addition, Argania fruit xyloglucan has no XUFG, a novel xyloglucan motif recently discovered in Argania leaf cell walls. Finally, the isolation and analysis of arabinogalactan-proteins showed that Argania fruit pulp is rich in these proteoglycans.

Uronic acids can be quantified either by a colorimetric determination after treatment with concentrated sulfuric acid and carbazole or by gas chromatography after methanolysis and subsequent acetylation. Both methods suffer from incomplete hydrolysis, an unavoidable degradation of the products to be analysed, and an inability to separate and quantify different types of uronic acids. In the present work, the fundamental chemistry involved in the two methods has been evaluated, and some modifications to increase their accuracy are suggested. By combining the two methods, a complete quantification of all individual types of uronic acids present in a sample can be achieved.

Normal phase-high performance liquid chromatography (NP-HPLC) coupled to matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF) tandem mass spectrometry is evaluated for the detailed structural characterization of various isomers of arabinoxylan (AX) oligosaccharides produced from endo-beta-(1-->4)-xylanase (endoxylanase) digestion of wheat AX. The fragmentation characteristics of these oligosaccharides upon MALDI-TOF/TOF high-energy collision induced dissociation (CID) were investigated using purified AX oligosaccharide standards labeled at the reducing end with 2-aminobenzoic acid (2-AA). A variety of cross-ring cleavages and 'elimination' ions in the fragment ion spectra provided extensive structural information, including Araf substitution patterns along the xylan backbone and comprehensive linkage assignment. The off-line coupling of this MALDI-CID technique to capillary normal phase HPLC enabled the separation and identification of isomeric oligosaccharides (DP 4-8) produced by endoxylanase digestion of AX. Furthermore, this technique was used to characterize structurally different isomeric AX oligosaccharides produced by endoxylanase enzymes with different substrate specificities.

Bilberries are known to have one of the most complex xyloglucan structures described in the plant kingdom until now. To characterise this structure, xyloglucans were enzymatically degraded and the oligosaccharides obtained were analysed. More than 20 different building blocks were found to make up the xyloglucan polymer. Bilberry xyloglucan was of XXXG-type, but some XXG-type oligomers were present, as well. The building blocks contain galactose-xylose (L) and fucose-galactose-xylose (F) side chains. In both side chains, the galactose units can be acetylated. In addition, beta-xylose-alpha-xylose (U) side chains were shown. This U chain was present in three building blocks described before (XUXG, XLUG and XUFG) and four novel blocks that have not been described in the literature previously: XUG, XUUG, XLUG and XXUG.

(Glucurono)arabinoxylans were extracted from barley husks and degraded with endo-beta-xylanase or subjected to periodate oxidation. The released oligosaccharide fragments were separated and isolated on Biogel-P2, and their structures were determined by NMR spectroscopy. The oligosaccharides identified consisted of beta-d-(1-->4)-linked xylopyranosyl residues, of which some were substituted at O-3 with alpha-l-arabinofuranosyl groups or at O-2 with 4-O-methylglucuronic acid. In addition to these substituents, a disaccharide side chain, 2-O-beta-d-xylopyranosyl-alpha-l-arabinofuranose, attached at position O-3 of the main chain, was proved to exist in arabinoxylan from barley husks. The compound was fully characterized with NMR, and all (1)H and (13)C NMR signals were assigned. The arabinose to xylose ratio was low (approximately 0.2) and no 2,3-disubstitution existed. No blocks of substituted xylose residues could be observed along the main chain.

Various arabino-xylo-oligosaccharides with known substitution patterns were assessed by negative ESI-Q-TOFMS and ESI-ITMS. The CID spectra of linear xylo-oligosaccharides and of nine isomeric mono- and disubstituted arabino-xylo-oligosaccharides established that structures differing in their substitution pattern can be differentiated by this approach. The negative-ion fragmentation spectra of the deprotonated quasi-molecular ions are mainly characterized by glycosidic cleavage ions from the C-series, which provide sequence informations, and by cross-ring cleavage (0,2)A(i) ions, which provide partial linkage information. When the collision energy increased, the cross-ring cleavage (0,2)A(i) ions underwent consecutive loss of water to produce (0,2)A(i)-18 fragment ions and glycosidic cleavage ions of the B-series are also produced besides the C(i) ions. Contrary to linear xylo-oligosaccharides, C(i) ions, which originate from C-3 monosubstituted xylosyl residues never produce the related cross-ring cleavage (0,2)A(i) ions. Disubstitution at O-2 and O-3 of xylosyl residues appears to enhance the production of the (0,2)A(i) ions compared to monosubstitution. For the differentiation of the mono- and disubstitution patterns of the penultimate xylosyl residue, the relative abundance of the glycosidic cleavage ions at m/z 263 and 299 found on Q-TOF CID spectra plays a relevant role and appears to be more informative than MS(n) spectra obtained on a ion trap instrument.

A set of arabinoxylan samples differing in their arabinose composition and various samples of arabino-xylo-oligosaccharide samples were analysed by Raman spectroscopy. Specific signatures for arabinose substitution were found in several spectral regions, that is, 400-600, 800-950 and 1030-1100 cm(-1). A linear relationship was observed between the peak ratio 855/895 cm(-1) of the second derivative spectra and the A/X ratio determined by chemical analysis. Moreover, spectral changes were observed in the 400-600 cm(-1) region assigned to the coupled vibrations mode in the skeleton: while the intensity of the band at 570 cm(-1) increased with the degree of substitution, that at 494 cm(-1) decreased. Similarly, a linear relationship was observed between the peak intensity ratio 570/494 cm(-1) calculated on the second derivative spectra and the composition data. Analysis of Raman spectra of arabino-xylo-oligosaccharides allowed to identify specific spectral features of disubstitution.

The present study provides a theoretical description of the different levels of structural organization that characterize the xylan polysaccharide in its native and hydrophobic lauroyl esterified forms. The goal of this study was to ascertain the role played by the hydroxyl or lauroyl side groups on the conformational flexibility of the xylan chain backbone. The results reported provide a detailed description of the low-energy conformers of the dimer segments, a complete characterization of the helical structures, an insight into the disordered state of the polysaccharide chains and an estimation of the cohesion of the amorphous solids. Esterification of xylan hydroxyl groups by lauric acid has a large effect on the conformational properties of the glycosidic bonds linking two repeat units. Both the location and the relative energies of the low energy areas of the potential energy surfaces strongly differ: extended and coiled conformations are preferred for the native and hydrophobic forms, respectively. Consequently, the predicted unperturbed polymer chain extension strongly depends on the structure, predicted Lp of the native xylan of 35 A compares favourably well with the experimental ones, this characteristic dramatically decreases to 9A for the hydrophobically modified chain. Curiously, only extended 2(1) and left-handed 3(1) helical structures are calculated stable for both polymers. The estimated cohesive parameters of amorphous bulks reveal that inter-chain interactions are stronger for the xylan chain than that for modified one, the former being stabilized by hydrogen bonds whereas hydrophobic interactions play a determinant role for the latter.

Molecular dynamics simulation was carried out on xyloglucan with explicit water molecules to investigate the folding mechanism of side chains onto a main chain in aqueous solution. The model xyloglucan was composed of 12 beta-D-glucopyranoses as a main chain substituted with six galactoses and three xyloses as side chains. Two conditions were set for the ribbon-like main chain; one is restricted to be 'flat' and the other is without restriction. The free main chain of xyloglucan has a 'twisted' conformation as the major one. Conformational folding of side chains onto the main chain was analyzed with dihedral angles at each glycosidic linkage. In a 5-ns calculation, the xyloglucan has a tendency to contract in both the restricted and the free systems, but the mode of contraction is different. Side chains tend to stick onto the flat surface of the main chain in the restricted system, while they do not tightly do so in the free one; instead the main chain takes a twisted and sometimes embowed conformation. This result indicates that the main chain has greater attractive forces to bind side chains when it is flat, while it loses the ability as it is twisted.

The degradation products of water-soluble wheat arabinoxylans treated with Aspergillus niger ferulic acid esterase (FAEA-able to cleave 5,5'- and 8-O-4'-ferulic acid dimers) have been characterised by atomic force microscopy (AFM) and size exclusion chromatography. The AFM images of arabinoxylans confirmed that a small proportion ( approximately 15%) of the population of arabinoxylan molecules contain xylan-based branches attached to the xylan-based backbone. Treatment with FAEA reduced the contour length of the molecules suggesting that certain dimeric ferulic acid linkages may play a previously unconfirmed role in the elongation of arabinoxylans. Overnight treatment with FAEA led to a reduction in the density of branches suggesting that they may also be linked to the backbone through phenolic linkages.

The structures of xyloglucans from several plants in the subclass Asteridae were examined to determine how their structures vary in different taxonomic orders. Xyloglucans, solubilized from plant cell walls by a sequential (enzymatic and chemical) extraction procedure, were isolated, and their structures were characterized by NMR spectroscopy and mass spectrometry. All campanulids examined, including Lactuca sativa (lettuce, order Asterales), Tenacetum ptarmiciflorum (dusty miller, order Asterales), and Daucus carota (carrot, order Apiales), produce typical xyloglucans that have an XXXG-type branching pattern and contain alpha-d-Xylp-, beta-D-Galp-(1-->2)-alpha-D-Xylp-, and alpha-L-Fucp-(1-->2)-beta-D-Galp-(1-->2)-alpha-D-Xylp- side chains. However, the lamiids produce atypical xyloglucans. For example, previous analyses showed that Capsicum annum (pepper) and Lycopersicon esculentum (tomato), two species in the order Solanales, and Olea europaea (olive, order Lamiales) produce xyloglucans that contain arabinosyl and galactosyl residues, but lack fucosyl residues. The XXGG-type xyloglucans produced by Solanaceous species are less branched than the XXXG-type xyloglucan produced by Olea europaea. This study shows that Ipomoea pupurea (morning glory, order Solanales), Ocimum basilicum (basil, order Lamiales), and Plantago major (plantain, order Lamiales) all produce xyloglucans that lack fucosyl residues and have an unusual XXGGG-type branching pattern in which the basic repeating core contains five glucose subunits in the backbone. Furthermore, Neruim oleander (order Gentianales) produces an XXXG-type xyloglucan that contains arabinosyl, galactosyl, and fucosyl residues. The appearance of this intermediate xyloglucan structure in oleander has implications regarding the evolutionary development of xyloglucan structure and its role in primary plant cell walls.

The relative activity of an endoxylanase towards water-unextractable (WU-AX) and water-extractable arabinoxylan (WE-AX) substrates, referred to as endoxylanase substrate selectivity, impacts the enzyme functionality in cereal-based biotechnological processes such as bread-making and gluten starch separation. A set of six endoxylanases representing a range of substrate selectivities as determined by a screening method using chromophoric substrates [Anal. Biochem.2003, 319, 73-77] was used to examine the impact of such selectivity on changes in structural characteristics of wheat WU-AX and WE-AX upon enzymic hydrolysis. While WE-AX degradation by the selected endoxylanases was very comparable with respect to apparent molecular mass (MM) profiles and arabinose to xylose ratio of the hydrolysates formed, WU-AX solubilisation and subsequent degradation of solubilised fragments gave rise to widely varying MM profiles, depending on the substrate selectivity of the enzymes. Enzymes with high selectivity towards WU-AX de facto generated higher MM fragments from WU-AX than enzymes with low selectivity. The arabinose to xylose ratios of solubilised fragments were independent of the degree of solubilisation.

Hemicellulose-type polysaccharides were isolated from the pericarp of seeds of Argania spinosa (L.) Skeels fruit by sequential alkaline extractions and fractionated by precipitation. Water soluble and water insoluble fractions were obtained, purified and characterized by sugar analysis and 1H and 13C NMR spectroscopy. The water soluble fractions were assumed to be (4-O-methyl-D-glucurono)-D-xylans, with 4-O-methyl-D-glucopyranosyluronic acid groups linked to C-2 of a (1-->4)-beta-D-xylan. The 1H NMR spectrum showed that the water soluble xylans have, on average, one non-reducing terminal residue of 4-O-methyl-D-glucuronic acid for every seven xylose units. The water insoluble fractions consisted of a neutral xylan with linear (1-->4)-beta-D-xylopyranosyl units.

The complex of xylan and iodine and its formation in a solution of xylan, CaCl2, and I2+KI was investigated by UV/Vis, second-derivative UV/Vis, and Raman spectroscopy. The complex forms only at very high concentrations of CaCl2, suggesting that when the water available in the solution is not sufficient to fully hydrate the calcium cation the chelation with the hydroxyl groups of the xylan can occur. The electronic spectra indicate that iodine is present in the form of three linear polyiodides I9(3-), I11(3-), and I13(3-) structures, which the Raman spectra show to be linear aggregates of the I3- and I5- substructures. Iodide concentration has a significant influence on the relative population of I9(3-), I11(3-), and I13(3-), as well as I3- and I5-, which lead to changes in both the UV/Vis absorption maxima shifts and changes in the Raman spectra. The key difference between this system of complexes with the linear polyiodide aggregates and that of amylose is that the longest aggregate observed with the amylose system, the I15(3-) polyanion, is not observed with the xylans. This indicates that the ordered arrays in the xylan-iodine complex do not exceed 4 nm in length. It is not possible to conclude at this time whether the ordered segment of the xylan molecule is linear or helical. If it is linear the length of the longest ordered arrays would be eight xylose residues. The number would exceed eight if the xylan molecule were helically wound.

This paper is the first multi-scale characterization of the xyloglucan extracted from seeds of the African tree Afzelia africana Se. Pers. It describes the extraction and characterization of this polysaccharide in terms of both primary monosaccharide and oligosaccharide composition. It also includes a study of the seed morphology. Morphological characterization includes optical, transmission, and scanning electron microscopy. The polysaccharide exists in thickened cell walls of the cotyledonary cells, and the extracted xyloglucan is structurally quite similar to those from tamarind seed and detarium. Nevertheless there are some subtle differences in the fine structure, particularly in the oligomeric xyloglucan composition. The chain flexibility of the polysaccharide is also discussed in the light of our recent measurements reported elsewhere [Biomacromolecules2004, 5, 2384-2391].