Perennial halophyte Salicornia neei Lag.: Cell wall composition and functional properties of its biopolymers.

Salicornia neei halophyte extends in Argentina seashores. To envisage potential applications, cell wall sequential extraction performed on dry plant yielded 1.1, 2.4, 0.3 and 0.9% of pectin fractions respectively extracted by room temperature water, 90 °C-water, CDTA and Na2CO3. They contained 21-33% uronic acids (UA) with low degree of methylation and 0.5-1.2 M ratios of neutral sugars to UA. High arabinose level suggests that long arabinan side-chains maintain cell wall flexibility in water deficit. Fractions also contained 10-36% of proteins. The KOH-soluble fractions (4.3%) were mainly arabinoxylans. At 2.0% w/v, pectin fractions developed "weak gel"-type networks with Ca2+, while arabinoxylans generated "dilute solutions". Cellulose (28%) and lignin (45.1%) were the main biopolymers in the final residue, which showed low water swelling capacity (3.6 mL/g) due to lignin, increasing when arabinoxylans were also present. Phenolics (9.8%) were mainly water-extractable. Salicornia is a source of biopolymers and antioxidants potentially useful for food applications.

A Comprehensive and Comparative Analysis of the Fucoidan Compositional Data Across the Phaeophyceae.

In the current review, compositional data on fucoidans extracted from more than hundred different species were surveyed through the available literature. The analysis of crude extracts, purified extracts or carefully isolated fractions is included in tabular form, discriminating the seaweed source by its taxonomical order (and sometimes the family). This survey was able to encounter some similarities between the different species, as well as some differences. Fractions which were obtained through anion-exchange chromatography or cationic detergent precipitation showed the best separation patterns: the fractions with low charge correspond mostly to highly heterogeneous fucoidans, containing (besides fucose) other monosaccharides like xylose, galactose, mannose, rhamnose, and glucuronic acid, and contain low-sulfate/high uronic acid proportions, whereas those with higher total charge usually contain mainly fucose, accompanied with variable proportions of galactose, are highly sulfated and show almost no uronic acids. The latter fractions are usually the most biologically active. Fractions containing intermediate proportions of both polysaccharides appear at middle ionic strengths. This pattern is common for all the orders of brown seaweeds, and most differences appear from the seaweed source (habitat, season), and from the diverse extraction, purification, and analytitcal methods. The Dictyotales appear to be the most atypical order, as usually large proportions of mannose and uronic acids appear, and thus they obscure the differences between the fractions with different charge. Within the family Alariaceae (order Laminariales), the presence of sulfated galactofucans with high galactose content (almost equal to that of fucose) is especially noteworthy.

Fucosylated chondroitin sulfate from the sea cucumber Hemioedema spectabilis: Structure and influence on cell adhesion and tubulogenesis.

Fucosylated chondroitin sulfate (FCS) HeSp was isolated from the Patagonian sea cucumber Hemioedema spectabilis. Ion-exchange chromatography was applied for purification of the FCS from the crude extract of sulfated polysaccharides. Analysis of monosaccharide and sulfate content of HeSp revealed the molar ratio of GlcA:GalNAc:Fuc:SO3Na as 1.15:1:1.1:3.9. Molecular weight of HeSp (44.1 kDa) was determined by GPC. According to the NMR spectral data, the main fragment of HeSp was the trisaccharide →3)-ß-d-GalNAc-(1→4)-ß-d-GlcA(3-O-α-l-Fuc)-(1→, where GalNAc units were sulfated either at O-4, at O-6 or both at O-4 and O-6. The fucosyl branches attached to O-3 of GlcA showed also different patterns of sulfation: Fucp2S4S, Fucp4S and Fucp3S4S were found in a ratio of 3.8:1.5:1. Besides, small amounts of the disaccharide fragment →3)-ß-d-GalNAc-(1→4)-ß-d-GlcA3S-(1→ were observed in a structure of HeSp. The polysaccharide was found to block cancer cells adhesion to platelet-coated surface and to inhibit tubulogenesis, thus demonstrating the potential antitumor activity.

Fucoidans from the phaeophyta Scytosiphon lomentaria: Chemical analysis and antiviral activity of the galactofucan component.

The brown seaweed Scytosiphon lomentaria produces moderate amounts of fucoidans. By cetrimide fractionation, typical heavily sulfated galactofucans are obtained, with no major signs of chemical heterogeneity, together with fractions with higher proportions of xylose, mannose and uronic acids. Anyway, fucose is the most important monosaccharide in most of the subfractions of the subsequent extracts. The fucan moieties appear to be mostly as 3-linked α-l-fucopyranosyl units, with several patterns of sulfate and branching. Galactose is mostly 6-linked, whereas mannose appears to be 2-linked, and xylose appears mostly as terminal stubs. Small amounts of 2-O-acetylated fucose units appear. A high and selective antiviral activity against HSV-1 and HSV-2 was determined for the galactofucan fractions whereas the uronofucoidans were inactive.

Husks of Zea mays as a potential source of biopolymers for food additives and materials' development.

Maize husks, an agricultural and industrial residue generated in a large volume, were investigated as a potential source of useful biopolymers. Thus, their chemical composition was firstly studied, after which two biopolymer products were obtained and characterized. Maize husks were dried and milled, obtaining a 210 µm-main particle size powder (MHP). It contained carotenes (4 mg/100 g), and exhibited antioxidant capacity (≈195 mg ascorbic acid/100 g MHP) coming also from extractable coumaric and cinnamic acids-derivatives (14 mg/100 g). A 31% of the MPH was water-soluble at room temperature, mainly constituted by fructose, glucose, and sorbitol of mesophylls' intracellular origin. The water insoluble fiber (WIF, ≈70%), which showed antioxidant capacity (≈25-33 mg ascorbic acid/100 g WIF), was almost entirely constituted by the cell wall biopolymers or alcohol insoluble residue (AIR) of the MPH, mostly arabinoxylans (≈26%) crosslinked by ferulic residues (18.6 mg/100 g MPH), and cellulose (26%). Low levels of pectins (5.5%) and lignin (7%) were found. Hence, a 1.25%-sulfur nanocellulose (NCC) was directly obtained with sulfuric acid (-15 mV Zeta-potential; 147 °C onset of thermal-degradation) without the necessity of previous delignification. On the other hand, a water soluble arabinoxylan enriched fraction (AX-EF) with pseudoplastic behavior in water and sensibility to calcium ions (≈3 Pa⋅s initial Newtonian-viscosity) was isolated by alkaline hydrolysis of diferulate bridges. Despite a 56% of crystallinity, NCC showed the highest water absorption capacity when compared to that of the AX-EF and AIR. Maize husks constitute an important source of biopolymers for development of materials and food additives/ingredients with relevant hydration and antioxidant properties.

Compositional changes in cell wall polysaccharides from five sweet cherry (Prunus avium L.) cultivars during on-tree ripening.

Excessive softening is a major cause of postharvest deterioration during transportation and storage of fresh cherries. In continuing our studies to identify the factors determining the textural differences between sweet cherry fruit genotypes, we evaluated the solubilization, depolymerization, and monosaccharide composition of pectin and hemicelluloses from five sweet cherry cultivars ('Chelan', 'Sumele', 'Brooks', 'Sunburst', and 'Regina') with contrasting firmness and cracking susceptibility at two developmental stages (immature and ripe). In contrast to what is usually shown in most fruits, cherry softening could occur is some cultivars without marked increases in water-soluble pectin. Although polyuronide and hemicellulose depolymerization was observed in the water-soluble and dilute-alkali-soluble fractions, only moderate association occurs between initial polymer size and cultivar firmness. In all the genotypes the Na2CO3-soluble polysaccharides (NSF) represented the most abundant and dynamic wall fraction during ripening. Firm cultivars showed upon ripening a lower neutral sugars/uronic acid ratio in the NSF, suggesting that they have a lower proportion of highly branched polyuronides. The similar molar ratios of arabinose plus galactose to rhamnose [(Ara+Gal)/Rha] suggest that the cultivars differed in their relative proportion of homogalacturonan (HG) and rhamnogalacturonan I (RG-I) rather than in the size of the RG side chains; with greater proportions of HG in firmer cherries. Ultraviolet matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was useful to identify the depolymerization patterns of weakly bound pectins, but gave less accurate results on ionically bound pectins, and was unable to find any pattern on covalently bound pectins.

The system of fucoidans from the brown seaweed Dictyota dichotoma: chemical analysis and antiviral activity.

Room-temperature acid (pH 2) extraction of Dictyota dichotoma thalli yielded 2.2% of sulfated polysaccharides. Further extraction with the same solvent at 70°C was conducted sequentially for nine times, with a total yield of 7.2%. Fucose was the main monosaccharide only in the room-temperature extract (EAR) and in the first 70°C extract (EAH1). The remaining fractions showed increasing amounts of mannose (the main neutral monosaccharide), xylose and uronic acids. Fractionation by means of cetrimide precipitation and redissolution in increasing sodium chloride solutions has allowed obtaining several subfractions from each extract. The fractions redissolved at lower NaCl concentrations have large amounts of uronic acids and lesser sulfate contents, whereas those redissolved at higher NaCl concentrations are heavily sulfated and have low uronic acid contents. For the fucose-rich extracts (EAR and EAH1), fractionation leads to uronoxylomannofucan-rich and galactofucan-rich fractions. The remaining extracts gave rise to complex mixtures, with mannose and uronic acid-rich polysaccharides. Moderate inhibitory effect against herpes virus (HSV-1) and Coxsackie virus (CVB3) were found for the galactofucan-rich fractions. Most of the other fractions were inactive against both viruses, although some xylomannan-rich fractions were also active against HSV-1.

Compositional changes in cell wall polysaccharides from apple fruit callus cultures modulated by different plant growth regulators.

The cell wall composition of apples callus cultures showed changes in the presence of 5 mg l(-1) of three different plant growth regulators (PGRs), namely picloram, abscisic acid and gibberellic acid. Although the structural functions of cell walls do not generally allow for pronounced variations of the total pectin and matrix glycan content, this work provides evidence that the addition of these plant growth regulators can rule, at least partly, cell wall metabolism in apple callus cultures. The chelator- and carbonate-extracts always had the analytical characteristics of pectins, with high proportions of uronic acids, arabinose and galactose as the main monosaccharides, and a significant proportion of rhamnose, but the cross-linking glycan fractions were still rich in RG-I-like material. The application of PGRs produced shifts of uronic acid and neutral sugars between fractions. Arabinose was the neutral sugar exhibiting more variations in apple callus cell wall. Picloram and abscisic acid produced an increase of the uronic acid contents of the cell walls. The AIRs obtained from calluses treated with different PGRs did not show large amounts of high molecular weight products, as determined by size-exclusion chromatography. For the carbonate-extract only the callus treated with picloram displayed two separated peaks for products of different molecular weights. The chromatographic profiles for the 4% KOH-extract displayed two peaks for all the treatments, one very sharp with high molecular weight, and another one wider of smaller molecular weight, whereas the difference between treatments can only be appraised through the areas of the peaks. This is the first report on cell wall composition from fruit calluses supplemented with different PGRs.

Effect of extraction time and temperature on the characteristics of loosely bound pectins from Japanese plum.

The cell wall composition of Japanese plums (Prunus salicina) at six developmental stages was previously evaluated (Ponce et al., J. Agric. Food Chem. 2010, 58, 2562-2570). This fruit is an interesting source of pectins, polysaccharides of valuable functionality for pharmaceutical and food formulations. In the present work it was investigated how the different conditions for the aqueous extraction of pectins from Japanese plums affect the yield as well as their chemical and rheological characteristics. It has been determined that extraction with water at room temperature for periods longer than 2h did not produce additional increment of yield (12%) but decreased the average molecular weights of the extracted pectins. Pectins with a degree of methylation ≈40% with high viscosity in water and with adequate molecular weights (≈72,000) were obtained. Conversely, utilization of boiling water for extraction increased considerably the yields (33-38%) but the extracted pectins showed significant lower viscosity in water in spite of their higher molecular weights. The poorer thickening ability was associated to the lower proportion of arabinose residues present in the hairy regions of the pectin macromolecules extracted by hot water, which led the polymers to interact more transiently in a 2% w/v water solution.

Compositional changes in 'Bartlett' pear ( Pyrus communis L.) cell wall polysaccharides as affected by sunlight conditions.

Preharvest conditions can have a great impact on fruit quality attributes and postharvest responses. Firmness is an important quality attribute in pear, and excessive softening increases susceptibility to bruising and decay, thus limiting fruit postharvest life. Textural characteristics of fruits are determined at least in part by cell wall structure and disassembly. Few studies have analyzed the influence of fruit preharvest environment in softening, cell wall composition, and degradation. In the current work 'Bartlett' pears grown either facing the sun (S) or in the shade (H) were harvested and stored for 13 days at 20 °C. An evaluation of fruit soluble solids, acidity, color, starch degradation, firmness, cell wall yield, pectin and matrix glycan solubilization, depolymerization, and monosaccharide composition was carried out. Sun-exposed pears showed more advanced color development and similar levels of starch degradation, sugars, and acids than shaded fruit. Sunlight-grown pears were at harvest firmer than shade-grown pears. Both fruit groups softened during storage at 20 °C, but even after ripening, sun-exposed pears remained firmer. Sunlight exposure did not have a great impact on pectin molecular weight. Instead, at harvest a higher proportion of water-solubilized uronic acids and alkali-solubilized neutral sugars and a larger mean molecular size of tightly bound glycans was found in sun-exposed pears. During ripening cell wall catabolism took place in both sun- and shade-grown pears, but pectin solubilization was clearly delayed in sun-exposed fruit. This was associated with decreased removal of RG I-arabinan side chains rather than with reduced depolymerization.

Characterization of acid-extracted pectin-enriched products obtained from red beet (Beta vulgaris L. var. conditiva) and butternut (Cucurbita moschata Duch ex Poiret).

Chemical and rheological characteristics of fractions enriched in soluble dietary fiber are reported. These fractions were obtained through acid hydrolysis of butternut (Cucurbita moschata Duch ex Poiret) and red beet (Beta vulgaris L. var. conditiva) cell wall enriched powders. Hydrolysis was performed using citric acid at different pH values and reaction times (2 and 3 h). Yields obtained for butternut fractions were between 21 and 28 g/100 g; for red beet, yields were 24 and 31 g/100 g for pH 1.5 and 11 and 17 g/100 g for pH 2.0 for previously mentioned times; in general, the increase of the yield was directly correlated with the decrease of pH and the increase of reaction time. Products enriched in low methoxyl pectins were obtained in all cases. At the lowest pH assayed, pectins were essentially constituted by homogalacturonan; a significant content of neutral sugars was determined at the higher extraction pH. Neutral sugars were constituted mainly by arabinose, galactose, rhamnose, and glucose in different proportions for each fraction; in general, butternut fractions showed high glucose contents. Flow behavior for 2.00% (w/v) aqueous systems of the different products was evaluated. Data obtained for fractions isolated at pH 1.5 fit to Herschel-Bulkley and Cross models while those isolated at pH 2.0 fit to Ostwald and Cross models. All samples showed low viscosity and, hence, poor thickening properties.

Compositional changes in cell wall polysaccharides from Japanese plum ( Prunus salicina Lindl.) during growth and on-tree ripening.

Climacteric Japanese plums were harvested at six developmental stages with no intermediate storage period, and cell wall compositional changes were analyzed. Arabinose proved to be the principal neutral monosaccharide constituent in cell walls during growth and the most dynamic neutral sugar in pectic fractions. Arabinose loss from tightly bound pectins was found to be a relatively early feature in the sequence of cell wall biochemical modifications, thus suggesting a softening-related role during Japanese plum on-tree ripening. Depolymerization of matrix glycans started at the end of the cell expansion phase and increased throughout ripening. Pectin solubilization was first detected during early ripening. Firmness loss did not correlate with polyuronide depolymerization early in ripening, but the last softening phase was associated with a strong depolymerization of cell wall polyuronides as well as a decrease in the arabinose/galactose ratio in loosely bound pectins. This is the first work that characterizes the temporal sequence of cell wall polysaccharide changes in Japanese plum.

Antiretroviral activity of fucoidans extracted from the brown seaweed Adenocystis utricularis.

Treatment of human immunodeficiency virus type 1 (HIV-1, causative agent of AIDS) infection represents a major challenge in antiviral therapeutics. Many difficulties are associated with the treatment, including toxicity, resistance and high costs. Taking this into account, research for novel compounds able to overcome these limitations is needed. Sulfated polysaccharides appear to be interesting, given their abundance as components of seaweeds. Herein, a series of fractions obtained from the brown seaweed Adenocystis utricularis was analysed for in vitro anti-HIV-1 activity. These fractions, which have anti-herpes simplex virus activity, were determined previously to belong to the family of fucoidans, sulfated polysaccharides obtained from the cell walls of brown seaweeds. Assays in human PBMC primary cell culture demonstrated that two of the five fractions analysed had potent anti-HIV-1 activity both against WT and drug-resistant HIV-1 strains. For active fractions, it was also shown that the inhibitory effect was not due to an inactivating effect on the viral particle (i.e. no virucidal activity was detected) but rather to a blockade of early events of viral replication. Given these encouraging results, these seaweed-derived fractions appear as good candidates for further studies on their potential for in vivo therapy and/or prophylaxis of HIV-1 infection.

Fucoidans from the brown seaweed Adenocystis utricularis: extraction methods, antiviral activity and structural studies.

The brown seaweed Adenocystis utricularis (family Adenocystaceae, order Ectocarpales sensu lato) was extracted in parallel with three solvents usually utilized for obtaining fucoidans: distilled water, 2% calcium chloride solution and diluted hydrochloric acid (pH 2) solution. In each case, the extraction was effected at room temperature and then at 70 degrees C. The extraction yields and characteristics of the products were similar in the three cases, with only minor differences. The analytical features of the products indicate that two different types of fucoidans are present in this seaweed. One of them, mostly extracted at room temperature, is composed mainly of L-fucose, D-galactose and ester sulfate (the 'galactofucan'). The other product (the 'uronofucoidan') is the major component of the extracts obtained at 70 degrees C. It is composed mainly of fucose, accompanied by other monosaccharides (mostly Man, but also Glc, Xyl, Rha and Gal), significant amounts of uronic acids and low proportions of sulfate ester. Fractionation with the cationic detergent cetrimide has allowed achieving a better separation of the galactofucan and uronofucoidan components. The galactofucans show a high inhibitory activity against herpes simplex virus 1 and 2, with no cytotoxicity, whereas the uronofucoidans carry no antiviral activity. Structural studies on the galactofucan fractions were carried out by methylation analysis, desulfation and NMR spectroscopy. The fucan constituent is mainly composed of 3-linked alpha-L-fucopyranosyl backbone, mostly sulfated at C-4, and branched at C-2 with non-sulfated fucofuranosyl and fucopyranosyl units, and 2-sulfated fucopyranosyl units. The galactan moiety is more heterogeneous, with predominant D-galactopyranose units linked on C-3 and C-6, and sulfation mostly on C-4, even in terminal non-reducing units. It may be inferred that at least some of these galactose units carry the alpha-configuration.