King oyster mushroom Pleurotus eryngii is cultivated worldwide for culinary and to improve human health. However, the potential of some Mediterranean representatives of this species is still not evaluated. This work focuses on the study of polysaccharides from fruiting bodies of two Tunisian strains, P. eryngii var. elaeoselini and P. eryngii var. ferulae, and, for comparison, one deposited P. eryngii originated from Korea. Polysaccharides were successively extracted with hot water using microwave heating and 1 mol L-1 aqueous sodium hydroxide. The crude hot water extracts were purified by treating them with proteolytic enzymes, and the alkaline extracts were purified by re-dissolving with dimethyl sulphoxide. In both cases, a decrease or removal of proteins was detected. Glucans predominated in all these products; the insoluble parts also contained chitin. The purified hot water extracts contained glycogen, β-d-glucans and mannogalactan. Branching (1 → 3)(1 → 6)-β-d-glucan was the major polysaccharide in the alkali-soluble fractions, while (1 → 3)-α-d-glucan was only a minor component. The Tunisian strains demonstrated a higher proportion of water-soluble polysaccharides, compared to the alkaline soluble ones, and more β-d-glucan in the insoluble chitin-glucan complexes. Fruiting body proteins of these strains are more available for solubilisation and enzymatic or alkaline degradation and, thus, may have higher nutritional value than those of the reference strain. As a source of proteins or polysaccharides, the Tunisian endemic P. eryngii strains of this study are promising for the domestication and cultivation of fruiting bodies for gastronomic purposes in the North African region.

• Klíčová slova
• Extraction, King oyster mushroom, Polysaccharides, Structure,
• MeSH
• druhová specificita MeSH
• fungální polysacharidy chemie   MeSH
• Pleurotus * chemie   MeSH
• plodnice hub * chemie   MeSH
• polysacharidy chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fungální polysacharidy MeSH
• polysacharidy MeSH

A set of fungal polysaccharide samples was characterised by elemental analysis and FTIR spectroscopy and compared with reference chitins, chitosans and β-D-glucans. The nitrogen to carbon (N/C) values and FTIR spectra were used to compare the samples based on their composition. It was found that the N/C ratio correlates well with deacetylation degree (DD) of chitosans and chitin/glucan ratio R(chit) of fungal chitin – β-D-glucan complexes with the exception of some samples having significant nitrogen and/or carbon admixtures. FTIR spectroscopy was indicative for the N-acetylation of chitins (chitosans) as well as for the chitin (chitosan) contribution to fungal polysaccharide preparations. Multivariate analyses of the FTIR data (HCA, PCA) discriminated samples and reference materials into several clusters depending on their similarity. Chitosan lactates, chitosan – β-D-glucans and chitin – β-D-glucans of high and low amounts of chitin were successfully discriminated from the reference polysaccharides and from each other. The proposed procedures based on the N/C ratio and multivariate analyses of FTIR spectra may be used in screening fungal polysaccharide preparations.

• Klíčová slova
• Chitin, FTIR, Fungal polysaccharides, N/C ratio, β-d-glucans,
• MeSH
• Aspergillus niger chemie   MeSH
• druhová specificita MeSH
• dusík chemie   MeSH
• fungální polysacharidy chemie   MeSH
• spektrofotometrie infračervená MeSH
• uhlík chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dusík MeSH
• fungální polysacharidy MeSH
• uhlík MeSH

Oyster mushrooms are an interesting source of biologically active glucans and other polysaccharides. This work is devoted to the isolation and structural characterization of polysaccharides from basidiocarps of the cultivated oyster mushroom, Pleurotus ostreatus. Five polysaccharidic fractions were obtained by subsequent extraction with cold water, hot water and two subsequent extractions with 1 m sodium hydroxide. Branched partially methoxylated mannogalactan and slightly branched (1→6)-β-d-glucan predominated in cold- and hot-water-soluble fractions, respectively. Alternatively, these polysaccharides were obtained by only hot water extraction and subsequent two-stage chromatographic separation. The alkali-soluble parts originating from the first alkali extraction were then fractionated by dissolution in dimethyl sulfoxide (DMSO). The polysaccharide insoluble in DMSO was identified as linear (1→3)-α-d-glucan, while branched (1→3)(1→6)-β-d-glucans were found to be soluble in DMSO. The second alkaline extract contained the mentioned branched β-d-glucan together with some proteins. Finally, the alkali insoluble part was a cell wall complex of chitin and β-d-glucans.

• Klíčová slova
• basidiocarps, fractionation, glucans, mannogalactan, oyster mushrooms, polysaccharides,
• MeSH
• chemická frakcionace MeSH
• chromatografie MeSH
• fungální polysacharidy chemie   izolace a purifikace   MeSH
• fytonutrienty chemie   izolace a purifikace   MeSH
• glukany chemie   MeSH
• molekulární struktura MeSH
• monosacharidy chemie   MeSH
• Pleurotus chemie   MeSH
• plodnice hub chemie   MeSH
• spektrální analýza MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fungální polysacharidy MeSH
• fytonutrienty MeSH
• glukany MeSH
• monosacharidy MeSH

The removal of noncovalently bound polysaccharide coating from the extracellular enzymes of Aspergillus niger, by the technique of compartmental electrophoresis, had a very dramatic effect on the stability of beta-glucosidase. The polysaccharide-beta-glucosidase complex was extremely resistant to proteinases and far more stable against urea and temperature as compared with polysaccharide-free beta-glucosidase. The beta-glucosidase-polysaccharide complex was 18-, 36-, 40- and 82-fold more stable against chymotrypsin, 3 mol/L urea, total thermal denaturation and irreversible thermal denaturation, respectively, as compared with polysaccharide-free beta-glucosidase. The activation energy of polysaccharide-complexed beta-glucosidase (55 kJ/mol) was lower than polysaccharide-free enzyme (61 kJ/mol), indicating a slight activation of the enzyme by the polysaccharide. No significant difference could be detected in the specificity constant (V/K(m)) for A-nitrophenyl beta-D-glucopyranoside between polysaccharide-free and polysaccharide-complexed beta-glucosidase. We suggest that the function of these polysaccharides secreted by fungi including A. niger might be to protect the extracellular enzymes from proteolytic degradation, hence increasing their life span.

• MeSH
• Aspergillus niger enzymologie   MeSH
• beta-glukosidasa chemie   izolace a purifikace   metabolismus   MeSH
• kinetika MeSH
• makromolekulární látky MeSH
• polysacharidy chemie   izolace a purifikace   MeSH
• stabilita enzymů MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• beta-glukosidasa MeSH
• makromolekulární látky MeSH
• polysacharidy MeSH

Electroinactive polysaccharides (PS) modified by osmium(VI) complexes with nitrogenous ligands produce redox couples at carbon and mercury electrodes. We show that PS adducts with Os(VI) 2,2'-bipyridine produce at ~-1.2 V (against Ag/AgCl/3 M KCl electrode) an additional peak at mercury and solid amalgam electrodes. This peak is due to the catalytic hydrogen evolution, allowing detection of PS (such as dextran and mannan) at picomolar concentrations.

• MeSH
• 2,2'-dipyridyl chemie   MeSH
• biosenzitivní techniky metody   MeSH
• dextrany analýza   MeSH
• elektrochemie metody   MeSH
• elektrody MeSH
• mannany analýza   MeSH
• osmium chemie   MeSH
• polysacharidy analýza   MeSH
• senzitivita a specificita MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 2,2'-dipyridyl MeSH
• dextrany MeSH
• mannany MeSH
• osmium MeSH
• polysacharidy MeSH

• Klíčová slova
• CORNEA *, EXPERIMENTAL LAB STUDY *, HYDROGEN-ION CONCENTRATION *, MACROMOLECULAR SYSTEMS *, MURAMIDASE *, POLYSACCHARIDES *, SWINE *,
• MeSH
• koncentrace vodíkových iontů * MeSH
• makromolekulární látky * MeSH
• muramidasa * MeSH
• polysacharidy * MeSH
• prasata MeSH
• rohovka * MeSH
• výzkum * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• makromolekulární látky * MeSH
• muramidasa * MeSH
• polysacharidy * MeSH

In this study, we focused on the isolation and structural characterization of polysaccharides from a basidiocarp of polypore fungus Ganoderma resinaceum. Polysaccharide fractions were obtained by successive extractions with cold water at room temperature (20 °C), hot water under reflux (100 °C), and a solution of 1 mol L-1 sodium hydroxide. The purity of all fractions was controlled mainly by Fourier transform infrared (FTIR) spectroscopy, and their composition and structure were characterized by organic elemental analysis; neutral sugar and methylation analyses by gas chromatography equipped with flame ionization detector (GC/FID) and mass spectrometry detector (GC/MS), respectively; and by correlation nuclear magnetic resonance (NMR) spectroscopy. The aqueous extracts contained two main polysaccharides identified as a branched O-2-β-d-mannosyl-(1→6)-α-d-galactan and a highly branched (1→3)(1→4)(1→6)-β-d-glucan. Mannogalactan predominated in the cold water extract, and β-d-glucan was the main product of the hot water extract. The hot water soluble fraction was further separated by preparative anion exchange chromatography into three sub-fractions; two of them were identified as branched β-d-glucans with a structure similar to the corresponding polysaccharide of the original fraction. The alkaline extract contained a linear (1→3)-α-d-glucan and a weakly branched (1→3)-β-d-glucan having terminal β-d-glucosyl residues attached to O-6 of the backbone. The insoluble part after all extractions was identified as a polysaccharide complex containing chitin and β-d-glucans.

• Klíčová slova
• Ganoderma resinaceum, polysaccharides, spectroscopy, wood decay fungi,
• Publikační typ
• časopisecké články MeSH

Glycosylation of proteins plays an important role in health and diseases. At present new simple and inexpensive methods of glycoprotein analysis are sought. We developed a monoclonal antibody Manost 2.1 in mice after immunization with the adduct of mannan with Os(VI)temed complex (temed is N,N,N',N'-tetramethylethylenediamine). The specificity of this antibody to different biomolecules treated with Os(VI)temed was tested using dot blot immunoassay. Manost 2.1 showed specificity toward Os(VI)temed-modified polysaccharides, glycoproteins and ribonucleotide at the 3'-end in DNA. The antibody recognized neither the unmodified compounds nor the non-glycosylated proteins treated with Os(VI)temed. We also performed western blotting and Coomassie silver blue staining of mixtures of biomacromolecules treated with Os(VI)temed and identified specifically the modified glycoproteins. The immunochemical method using Manost 2.1 was compared with electrochemical analyses based on redox signals of the Os(VI)temed adducts, with similar results in terms of sensitivity. This new antibody-based approach opens the door for rapid and inexpensive analysis of glycans and glycoproteins in various scientific and medical fields, including cancer research and the future application of glycoprotein detection in clinical practice.

• Klíčová slova
• Antibody against chemically modified carbohydrates, Carbohydrate chemical modification, Carbon and mercury electrodes, Glycoprotein adduct immunoassays, Os(VI) complexes, Voltammetry of chemically modified glycoproteins,
• MeSH
• DNA MeSH
• glykoproteiny analýza   MeSH
• imunoanalýza * MeSH
• monoklonální protilátky MeSH
• myši MeSH
• nukleové kyseliny chemie   MeSH
• polysacharidy analýza   MeSH
• ribosa analýza   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA MeSH
• glykoproteiny MeSH
• monoklonální protilátky MeSH
• nukleové kyseliny MeSH
• polysacharidy MeSH
• ribosa MeSH

ETHNOPHARMACOLOGICAL RELEVANCE: Geranium sanguineum L. is used for treatment of inflammations, anemia, malignant diseases of the blood-forming organs, diarrhea, respiratory infections, etc. Only flavonoids in root extracts have been elucidated as immunostimulating and anti-inflammatory compounds, and polysaccharides in the herb have not been examined. AIM OF THE STUDY: to compare the chemical features of polysaccharide complexes (PSCs) from leaves (GSL-PSC) and roots (GSR-PSC) of G. sanguineum, as well as their immunomodulatory activities on leukocytes after inflammation, and effects on the growth of different bacteria. MATERIALS AND METHODS: The samples were isolated by water extraction and their structural features were studied by 2D NMR spectroscopy. The stimulatory effects of both PSCs on human leukocytes were analyzed with flow cytometry. Their suppressive activities on the oxidative burst in blood and derived neutrophils against opsonized zymosan and phorbol myristate acetate were investigated. The effects of the samples on viability, NO and interleukin 6 (IL-6) syntheses in RAW264.7 cells after inflammation with lipopolysaccharides (LPS) were tested. The prebiotic and anti-biofilm activities of the PSCs were evaluated. RESULTS: The total carbohydrate content in the samples was significant (73.6-76.8%). GSL-PSC contained pectins, which were rich in homogalacturonan (HG), and smaller amounts of rhamnogalacturonan (RG) type I, decorated by 1,5-α-L-Araf, 1,4- and 1,6-β-D-Galp chains. GSR-PSC contained starch, followed by pectins with lower HG content and more RG-I regions, substituted by 1 → 3,5-α-L-arabinans and 1 → 3,6-β-D-galactans. GSL-PSC and GSR-PSC (200 μg/mL) increased monocyte and granulocyte cell counts, but GSR-PSC also elevated T helper and B cell levels in a normal and activated state. GSR-PSC triggered a dose-dependent (50-200 μg/mL) oxidative burst in blood, but alleviated it after inflammation even in blood-derived neutrophils. It was free of LPS, and activated NO and IL-6 productions in RAW264.7 cells better than GSL-PSC, without affecting their viability. Both PSCs (2.0%, w/v) stimulated probiotic co-cultures between Clostridium beijerinckii strains and Lactobacillus sp. ZK9, and inhibited the growth and biofilm formation of Escherichia coli, Streptococcus mutans and Salmonella enterica. CONCLUSIONS: The PSs in G. sanguineum could be involved in the stimulatory effects on blood-forming organs and anti-inflammatory action of aqueous root extracts in case of infections. These PSs should be included in synbiotic foods to support the treatment of inflammations and infections in the gut.

• Klíčová slova
• Geranium sanguineum, Immunomodulatory activity, Inflammation, Interleukin 6, Lactose (PubChem CID: 6134), Lipopolysaccharide, Nitric oxide (PubChem CID: 145068), Pectic polysaccharide, Phorbol 12-myristate 13-acetate (PubChem CID: 27924), Polysaccharides, Probiotic bacteria, Starch, Structural analysis, Zymosan,
• MeSH
• antiflogistika MeSH
• Geranium * chemie   MeSH
• interleukin-6 MeSH
• lidé MeSH
• lipopolysacharidy MeSH
• myši MeSH
• pektiny farmakologie   MeSH
• polysacharidy * farmakologie   MeSH
• RAW 264.7 buňky MeSH
• zánět farmakoterapie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika MeSH
• interleukin-6 MeSH
• lipopolysacharidy MeSH
• pektiny MeSH
• polysacharidy * MeSH

Hot water extract from biomass of heterotrophic mutant green alga Parachlorella kessleri HY1 (Chlorellaceae) was deproteinised, and three polysaccharidic fractions were obtained by preparative chromatography. The low-molecular fraction (1.5 × 104g mol-1) was defined mainly as branched O-2-β-xylo-(1→3)-β-galactofuranan where xylose is partially methylated at O-4. Two high-molecular fractions (3.05 × 105 and 9.84 × 104g mol-1) were complex polysaccharides containing α-l-rhamnan and xylogalactofuranan parts in different ratios. The polysaccharides were well soluble in hot water and, upon cooling, tended to self-segregate. Immunomodulatory activities of the obtained fractions were preliminary tested using ELISA, FACS and ImmunoSpot kits. The polysaccharides increased the TNF-α production in melanoma bearing mice with much higher intensity than in healthy mice. This was in agreement with the FACS results on T and B cells indicating their possibly secondary activation by innate immunity cells.

• Klíčová slova
• Aggregation, Chlorella, Immunomodulatory activity, Rhamnans, Structure, Xylogalactofuranan,
• MeSH
• B-lymfocyty účinky léků   imunologie   patologie   MeSH
• CD antigeny genetika   imunologie   MeSH
• Chlorophyta chemie   MeSH
• imunologické faktory chemie   izolace a purifikace   farmakologie   MeSH
• interferon gama genetika   imunologie   MeSH
• interleukin-2 genetika   imunologie   MeSH
• interleukin-4 genetika   imunologie   MeSH
• lipopolysacharidy antagonisté a inhibitory   farmakologie   MeSH
• melanom imunologie   patologie   MeSH
• metylace MeSH
• molekulová hmotnost MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory kůže imunologie   patologie   MeSH
• polysacharidy chemie   izolace a purifikace   farmakologie   MeSH
• primární buněčná kultura MeSH
• regulace genové exprese účinky léků   MeSH
• rostlinné extrakty chemie   MeSH
• rozpustnost MeSH
• sacharidové sekvence MeSH
• T-lymfocyty účinky léků   imunologie   patologie   MeSH
• TNF-alfa genetika   imunologie   MeSH
• voda MeSH
• xylosa chemie   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• CD antigeny MeSH
• Il4 protein, mouse MeSH Prohlížeč
• imunologické faktory MeSH
• interferon gama MeSH
• interleukin-2 MeSH
• interleukin-4 MeSH
• lipopolysacharidy MeSH
• polysacharidy MeSH
• rostlinné extrakty MeSH
• TNF-alfa MeSH
• voda MeSH
• xylosa MeSH