Structural properties of the extracellular biopolymer (ß-D-xylo-α-D-mannan) produced by the green microalga Gloeocystis vesiculosa Nägeli.

Many species of microalgae produce a relatively diverse range of metabolites that are interesting for biotechnological applications, and among them exopolysaccharides attract attention due to their structural complexity, biological activities, biodegradability or biocompatibility. An exopolysaccharide of high molecular weight (Mp) of 6.8 × 105 g/mol was obtained by cultivation of the freshwater green coccal microalga Gloeocystis vesiculosa Nägeli 1849 (Chlorophyta). Chemical analyses revealed a dominance of Manp (63.4 wt%), Xylp and its 3-O-Me-derivative (22.4 wt%), and Glcp (11.5 wt%) residues. The results of the chemical and NMR analyses showed an alternating branched 1,2- and 1,3-linked α-D-Manp backbone terminated by a single ß-D-Xylp and its 3-O-methyl derivative at O2 of the 1,3-linked α-D-Manp residues. The α-D-Glcp residues were found mainly as 1,4-linked and to a lesser extent as the terminal sugar, indicating partial contamination of ß-D-xylo-α-D-mannan with amylose (∼10 wt%) in G. vesiculosa exopolysaccharide.

An arabinan isolated from the antioxidant active fraction of wild blackthorn fruits (Prunus spinosa L.).

Despite the many health benefits of wild blackthorn fruits, their use is negligible and little explored. Preliminary results suggest that this fruit is a rich source of natural antioxidants. Antioxidant active fraction, extracted from wild blackthorn fruits by cold water extraction (Cw), was subjected to ion-exchange chromatography, which gave ten fractions after a successive elution with water, sodium chloride and sodium hydroxide solutions. Fraction eluted by water contained an arabinan of a low molecular weight (Mw = 5000). Its chemical and spectroscopic studies have revealed a highly branched structure with backbone composed of 1,5-linked α-l-arabinofuranose residues; about 76% of them are branched dominantly through O3 and much less through O2, however, some arabinose units (∼8%) were fully branched. In addition to terminal α-L-Araf, the presence of ß-L-Araf unit was also identified in blackthorn L-arabinan. Antioxidant activity tests of Cw ion exchange fractions revealed significant effects only of fractions eluted with 1 M sodium chloride (6F) and sodium hydroxide (7F) solutions, while phenolic free arabinan did not possess this effect. Phenolic compounds linked via α-l-arabinosyl side chains appear to be responsible for the antioxidant effect of two Cw fractions.

Isolation, chemical characterization and antioxidant activity of Prunus spinosa L. fruit phenolic polysaccharide-proteins.

Wild blackthorn fruits are a slightly explored area and their use is negligible, although they show many therapeutic effects. Polysaccharide complexes were solubilized from ripe Prunus spinosa L. fruits by sequential extraction with water (cold and hot), oxalate, ammonia, and sodium hydroxide solutions. The extracted fractions were rich in carbohydrates, phenolics, and proteins, indicating the presence of complexes of these compounds. It was found that most of the material was released with 5% KOH and hot water. The aqueous fractions had a high molecular weight and also polydispersity index, while the other fractions had low molecular weight. Based on the monosaccharide composition, three groups of fractions were observed. The aqueous and oxalate fractions represent highly esterified pectin material, ammonia extraction provided a mixture of de-esterified pectin and hemicelluloses, and alkaline isolates were rich in arabinogalactan and hemicelluloses. Antioxidant activity tests revealed significant effects of blackthorn samples, alkali-extracted fractions at concentrations of 0.02 and 0.05 mg/mL achieved DPPH radical scavenging comparable to that of vitamin C. Most samples achieved maximum activity comparable to vitamin C at a concentration of 0.1 mg/mL. An analogous effect was also found by the FRAP test, all fractions had a significant reducing capacity at a concentration of 0.05 mg/mL, but only the alkaline fractions reached almost vitamin C level. Preliminary results may support interest in these "undiscovered" natural fruits as a rich source of natural antioxidants.

Lactylated acidic exopolysaccharide produced by the cyanobacterium Nostoc cf. linckia.

Cyanobacteria produce a wide range of metabolites of interest for industrial or medical use. The cultivation of freshwater Nostoc cf. linckia yielded 5.4 g/L of a crude exopolysaccharide (cEPS) with a molecular weight of 1.31 × 105 g/mol. Ion-exchange chromatography of cEPS yielded two dominant fractions, EPS-1 and EPS-2, differing in molecular weight. The lower molecular weight fraction (EPS-1) was subjected to structural studies. Results of chemical and spectroscopic analyses showed that three of the four dominant sugars, glucose, galactose and xylose are 1,4-linked in the backbone in the following order: [→4)-ß-D-Xylp-(1 â†’ 4)-ß-D-Glcp-(1 â†’ 4)-α-D-Galp-(1 â†’ 4)-ß-D-Glcp-(1→]n. Terminal mannose residues were identified as side chains linked at C3 of every third backbone xylose and every second glucose is branched at C6 by 3-O-lactyl-ß-D-glucuronic acid (nosturonic acid). Antioxidant properties of EPS were tested using two in vitro methods. Both assays showed that the cEPS was more active than purified EPS-1 and EPS-2 fractions and deproteinized EPS.

Structural characterisation of polysaccharides from roasted hazelnut skins.

Two polysaccharide fractions sequentially extracted with water 1W and alkali 1A, were isolated from the hazelnut skins. The monosaccharide composition together with the FTIR and NMR analyses, indicated that both fractions are formed from a mixture of polysaccharides. The fraction 1W consists of methyl-esterified pectic polysaccharide with rhamnogalacturonan I blocks, branching with arabinose side chains, and with 1,5-, 1,3,5-arabinan and galactan polysaccharides. The fraction 1A is a mixture of deesterified rhamnogalacturonan I and 1,5-, 1,3,5-arabinan and 4-O-Me-glucuronoxylan polysaccharides. The presence of unsaturated galacturonic acid and the heterogeneity of the molecular weights, which Mw ranged between 3.6 and 39 kg mol-1, indicated the pectin degradation during roasting.

(3, 2)D 1H, 13C BIRDr,X-HSQC-TOCSY for NMR structure elucidation of mixtures: application to complex carbohydrates.

Overlap of NMR signals is the major cause of difficulties associated with NMR structure elucidation of molecules contained in complex mixtures. A 2D homonuclear correlation spectroscopy in particular suffers from low dispersion of 1H chemical shifts; larger dispersion of 13C chemical shifts is often used to reduce this overlap, while still providing the proton-proton correlation information e.g. in the form of a 2D 1H, 13C HSQC-TOCSY experiment. For this methodology to work, 13C chemical shift must be resolved. In case of 13C chemical shifts overlap, 1H chemical shifts can be used to achieve the desired resolution. The proposed (3, 2)D 1H, 13C BIRDr,X-HSQC-TOCSY experiment achieves this while preserving singlet character of cross peaks in the F1 dimension. The required high-resolution in the 13C dimension is thus retained, while the cross peak overlap occurring in a regular HSQC-TOCSY experiment is eliminated. The method is illustrated on the analysis of a complex carbohydrate mixture obtained by depolymerisation of a fucosylated chondroitin sulfate isolated from the body wall of the sea cucumber Holothuria forskali.

Description of the male of Acentrus boroveci Kostál, 2014 (Coleoptera: Curculionidae: Acentrusini).

The male of the recently described Acentrus boroveci Kostál, 2014 is described for the first time, and characters to differentiate it from males of the two other species of Acentrus Desmarest, 1839 are given.

Bioactive hemicelluloses alkali-extracted from Fallopia sachalinensis leaves.

Fallopia sachalinensis, regarded as an invasive plant in Europe and designated for disposal, is traditionally used in Japan and China as herbal medicine. Attempted for valorization of the leaves, this paper reports on two protein-free polysaccharide fractions, a neutral (FS-5A) and an acidic (FS-5B) one, obtained via alkali extraction and consecutive purification. Both fractions were characterized by chemical, molecular, structural and bioactive properties. FTIR and 1D/2D NMR analyses revealed that FS-5A consisted of a fucogalactoxyloglucan, whereas, glucuronoxylan was the major hemicellulose in FS-5B accompanied with low proportions of fucosylated xyloglucan and pectic RG-I. Both hemicellulose fractions exhibited significant immunostimulating activity in the complement-fixation test and the latter had noticeable DPPH radical-scavenging ability. The results completed information about neutral and acidic bioactive polysaccharide components present in the leaves of F. sachalinensis.

Non-cellulosic polysaccharides from the leaves of small balsam (Impatiens parviflora DC.).

The leaves of the annual plant Impatiens parviflora DC., a herbal medicine in Asian countries and invasive in managed forests and natural environments in Central Europe, have the potential as a source of bioactive phenolic compounds and polysaccharides. Extractives accounted for ∼22% of the leaves, whereby, the methanolic extract contains mainly caffeic acid, ferulic acid, kaempferol, and quercetin derivatives, and 1,2,4-trihydroxynaphthalene-1-O-glucoside. From the pre-extracted leaves, non-cellulosic polysaccharides were isolated by a five-step extraction procedure using as extractants cold water, 0.05 M EDTA, and DMSO in the first three steps, and 1% and 5% NaOH in the last two steps. The isolated polysaccharide fractions were characterized by chemical, physicochemical, and spectroscopic analyses (FTIR and (1)H NMR). The first three fractions contained mainly pectin and the alkali-extracted ones methylglucuronoxylan and arabinogalactan. The suggested structural features were confirmed using HSQC NMR and COSY experiments for polysaccharides of the EDTA-fraction and the Pronase-treated 5% NaOH-fraction. The EDTA fraction comprised a pectin with low degree of methyl-esterification (DM, 37%) with few rhamnogalacturonan RG I segments bearing ß-1,4-galactan as side chains. The alkali-extracted fraction comprised a degraded methylglucuronoxylan and type II 3,6-arabinogalactan in about equal amounts.

Structural features of two heteroxylan polysaccharide fractions from wheat bran with anti-complementary and antioxidant activities.

Wheat bran is a rich source of bioactive substances ascribed to its arabinoxylan component. Two water-soluble arabinoxylans were sequentially extracted from wheat bran. WB1, released during enzymatic digestion of starch and protein, contained medium-branched arabinoxylan (A/X=0.88) consisting of 3-O-substituted (22%), di-substituted (19.8%) and 58% unsubstituted Xylp residues. It was slightly contaminated with (1→3,1→4)-ß-glucan and arabinogalactan, and free of protein. WB2 extracted with 0.5% NaOH contained ∼95% arabinoxylan (A/X=1.09). WB2 and two 5% NaOH-extracted arabinoxylans were rich in protein and phenolic compounds. All radical-scavenging assays indicated a relation with the protein and total phenolics contents. The protein-free WB1 displayed the highest hydroxyl radical scavenging effect indicating the distinct role of phenolic acids. The immunomodulatory activity of WB1 was somewhat lower, whereas, that of WB2 higher in comparison to the immunogenic polysaccharide PMII. The arabinoxylans have the potential as immuno-enhancing and antioxidant additives in functional foods.

Structural diversity of pectins isolated from the Styrian oil-pumpkin (Cucurbita pepo var. styriaca) fruit.

To evaluate the seeded fruit biomass of the Styrian oil-pumpkin in view of its pectin component, a series of acidic polysaccharides were isolated by a six-step sequential extraction using hot water, EDTA, dilute HCl (twice) and dilute and stronger NaOH solutions. Chemical, physicochemical and spectroscopy analyses revealed that the first four fractions comprised partially methyl-esterified and acetylated pectins with varying proportions of rhamnogalacturonan regions ramified with galactose- and arabinose-containing side chains and showed considerable polymolecularity. The alkali-extracted polysaccharides contained lower amounts of pectins with homogalacturonan and arabinose-rich rhamnogalacturonan regions next to hemicelluloses prevailing in the last polysaccharide. Using (1)H-(13)C HSQC and HMBC spectroscopy, the resonances of free and methylesterified galacturonic acid residues in the purified acid-extracted pectin were unambiguously established and various diads formed by both residues identified. The results might serve as a basis for searching technological conditions to produce pectin from the oil-pumpkin fruit biomass.

Suppressive effect of pectic polysaccharides from Cucurbita pepo L. var. Styriaca on citric acid-induced cough reflex in guinea pigs.

Several water-soluble pectic polysaccharides were isolated from the pumpkin fruit biomass and characterized by composition, structural features and molecular properties. The pectic polysaccharides were tested for antitussive activity by studying the effects of citric acid-induced cough reflex in guinea pigs and reactivity of the airway smooth muscle in vivo conditions in comparison to the narcotic drug codeine. Oral administration of all pectic polysaccharides from pumpkin inhibited the number of coughs induced by citric acid in guinea pigs, but to various extents. The results indicated that the antitussive activity of the pectic polysaccharides is affected by their molecular and structural properties, whereby a synergistic action between the polysaccharide and non-carbohydrate components on the biological response has been suggested as well. The cough depressive efficacy of most of the tested polysaccharides was comparable and even higher than that of codeine. Moreover, the application of these polysaccharides provoked any side effects what is their advantage towards the conventional opioid-derived antitussive agents.