Dictyosphaerium chlorelloides is a green microalga from the Chlorella clade that produces highly viscous exocellular polysaccharides. The cell wall polysaccharides of this alga have not been studied in detail. In this article, water-soluble polysaccharides from D. chlorelloides biomass were extracted with hot water and purified by preparative chromatography. The composition, structural features and molecular masses of subsequently eluted fractions F1, F2, F3, F4 and F5 (minor) were determined. Three high-yield products F1, F3 and F4 consisted mainly of galactopyranosyl, 2-O-methyl-galactopyranosyl, rhamnopyranosyl and mannopyranosyl units at different proportions, while F2 was rich in glucose. Immunoactivity of these fractions was evidenced in a mixed population of immune cells derived from mice spleens after incubation with polysaccharides by flow cytometry, MTT and Immunospot assays. These fractions, except F2, demonstrated selective immunostimulant activity, and the F1 fraction induced the most potent effect, closely followed by the F3 and F4 fractions. The in vivo mechanism of their action is associated with the activation of innate immunity and shapes the immune response to the Th1 type.
- MeSH
- adjuvancia imunologická farmakologie chemie izolace a purifikace MeSH
- buněčná stěna * chemie MeSH
- Chlorophyta chemie MeSH
- mikrořasy * chemie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- polysacharidy * farmakologie chemie izolace a purifikace MeSH
- slezina cytologie účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Galactooligosaccharides (GOS) are lactose-derived functional ingredients applied in food products and have great potential in health protection. The conversion of lactose to GOS commonly occurs using β-galactosidases of mould, yeast and bacterial origin. The yield and structure of the resulting GOS depend on the enzyme used and the reaction conditions. This work focuses on the structural analysis of the products obtained with four commercial β-galactosidases Maxilact LGI 5000 (ML), Maxilact A4 MG (MA), Saphera 2600 L (SA) and NOLA Fit 5500 (NL) to evaluate their efficiency and specificity. HPLC, ESI-MS and NMR spectroscopy were applied to characterise the GOS preparations. GOS were separated from the reaction mixture using activated charcoal treatment. HPLC analysis confirmed that most of the monosaccharides and a part of the lactose, but also some other disaccharides, probably allolactose and 6-galactobiose, were retained by charcoal. In all the products, ESI-MS analysis detects oligosaccharides up to hexamers. NMR spectra confirmed the presence of GOS of various configurations and polymerisation degrees and evaluated the specificity of used enzymes. MA preferably forms 1,6- and 1,4-glycosidic bonds, and bacterial enzymes NL and SA also form 1,2- and 1,3- glycosidic bonds, while yeast enzyme ML cannot produce new 1,4-glycosidic bonds. The mould enzyme MA showed the highest trans-galactosylation activity, forming longer GOS oligomers than the other enzymes.
- MeSH
- beta-galaktosidasa * metabolismus chemie MeSH
- galaktosa chemie metabolismus MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací * metody MeSH
- laktosa metabolismus chemie MeSH
- magnetická rezonanční spektroskopie * metody MeSH
- oligosacharidy * chemie metabolismus MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
The ear- to shell-shaped fruiting bodies of the genus Auricularia are widely used as food and in traditional medicinal remedies. This study was primarily focused on the composition, properties and potential use of the gel-forming extract from Auricularia heimuer. The dried extract contained 50% soluble homo- and heteropolysaccharides, which were mainly composed of mannose and glucose, acetyl residues, glucuronic acid and a small amount of xylose, galactose, glucosamine, fucose, arabinose and rhamnose. The minerals observed in the extract included approximately 70% potassium followed by calcium. Among the fatty and amino acids, 60% unsaturated fatty acids and 35% essential amino acids could be calculated. At both acidic (pH 4) and alkaline (pH 10) conditions, the thickness of the 5 mg/mL extract did not change in a temperature range from -24 °C to room temperature, but decreased statistically significantly after storage at elevated temperature. At neutral pH, the studied extract demonstrated good thermal and storage stability, as well as a moisture retention capacity comparable to the high molecular weight sodium hyaluronate, a well-known moisturizer. Hydrocolloids that can be sustainably produced from Auricularia fruiting bodies offer great application potential in the food and cosmetic industries.
- Publikační typ
- časopisecké články MeSH
Vibrational spectroscopic methods are widely used in the molecular diagnostics of carcinogenesis. Collagen, a component of connective tissue, plays a special role as a biochemical marker of pathological changes in tissues. The vibrational bands of collagens are very promising to distinguish between normal colon tissue, benign and malignant colon polyps. Differences in these bands indicate changes in the amount, structure, conformation and the ratio between the individual structural forms (subtypes) of this protein. The screening of specific collagen markers of colorectal carcinogenesis was carried out based on the FTIR and Raman (λex 785 nm) spectra of colon tissue samples and purified human collagens. It was found that individual types of human collagens showed significant differences in their vibrational spectra, and specific spectral markers were found for them. These collagen bands were assigned to specific vibrations in the polypeptide backbone, amino acid side chains and carbohydrate moieties. The corresponding spectral regions for colon tissues and colon polyps were investigated for the contribution of collagen vibrations. Mentioned spectral differences in collagen spectroscopic markers could be of interest for early ex vivo diagnosis of colorectal carcinoma if combine vibrational spectroscopy and colonoscopy.
- MeSH
- kolagen MeSH
- kolonoskopie * metody MeSH
- kolorektální nádory * diagnóza MeSH
- lidé MeSH
- Ramanova spektroskopie metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Within the group of higher fungi, edible medicinal mushrooms have a long history of being used as food and in folk medicine. These species contain biologically active substances with many potential beneficial effects on human health. The Pleurotus genus is representative of medicinal mushrooms because Pleurotus ostreatus is one of the most commonly cultivated culinary mushrooms. In our study, we focused on lesser-known species in the genus Pleurotus and measured their antioxidant and anti-inflammatory activity. We prepared extracts of the mushrooms and analyzed them using HPLC-HRMS, GC-MS, and 1H-NMR. Significant differences in biological activities were found among the Pleurotus spp. extracts. A MeOH extract of P. flabellatus was the most active as a radical scavenger with the highest ORAC, while a chloroform extract had significant anti-inflammatory COX-2 activity. The 80% MeOH extract of P. flabellatus contained the highest amounts of ergosterol, ergothioneine, and mannitol. The 80% MeOH extract of P. ostreatus Florida was the most active in the NF-κB inhibition assay and had the highest content of β-glucans (43.3% by dry weight). Given the antioxidant and anti-inflammatory properties of P. flabellatus, the potential therapeutic usefulness of this species is worth evaluating through in-depth investigations and confirmation by clinical trials.
- Publikační typ
- časopisecké články MeSH
Excessive osteoclast differentiation and/or bone resorptive function causes a gradual loss of bone, leading to the pathogenesis of bone diseases such as osteoporosis (OP). In this study, a sulfated glucuronorhamnoxylan polysaccharide (designated SPS-CF) of the green alga Capsosiphon fulvescens was evaluated for anti-osteoporotic activity using osteoclastic cells differentiated from RAW264.7 macrophages by receptor activator of NF-κB ligand (RANKL) treatment and ovariectomized (OVX) female mice as a postmenopausal OP model. With negligible cytotoxicity, SPS-CF (50 μg/mL) significantly suppressed tartrate-resistant acid phosphatase (TRAP) activity, actin ring formation, and expression of matrix metalloproteinase 9 (MMP-9), cathepsin K, TRAF6, p-Pyk2, c-Cbl, c-Src, gelsolin, carbonic anhydrase II (CA II), and integrin β3, indicating that SPS-CF inhibits the differentiation and bone resorptive function of osteoclasts. Removal of sulfate groups from SPS-CF abolished its anti-osteoclastogenic activities, demonstrating that sulfate groups are critical for its activity. Oral administration of SPS-CF (400 mg/kg/day) to OVX mice significantly augmented the bone mineral density (BMD) and serum osteoprotegerin (OPG)/RANKL ratio. These results demonstrated that SPS-CF exerts significant anti-osteoporotic activity by dampening osteoclastogenesis and bone resorption via downregulation of TRAF6-c-Src-Pyk2-c-Cbl-gelsolin signaling and augmentation of serum OPG/RANKL ratios in OVX mice, suggesting that SPS-CF can be a novel anti-osteoporotic compound for treating postmenopausal OP.
- MeSH
- buněčná diferenciace MeSH
- Chlorophyta * metabolismus MeSH
- faktor 6 asociovaný s receptory TNF metabolismus MeSH
- fokální adhezní kinasa 2 metabolismus MeSH
- gelsolin metabolismus MeSH
- kyselá fosfatasa rezistentní k tartarátu metabolismus MeSH
- myši MeSH
- NF-kappa B metabolismus MeSH
- osteoporóza * farmakoterapie MeSH
- resorpce kosti * farmakoterapie MeSH
- sírany metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Chitosan and β-glucan are substances that are very widely used in the pharmaceutical and food industries, medicine and other areas. These polysaccharides have immense significance in human metabolism. They are able to affect the levels of cholesterol and lipids. Chemical modification of these polysaccharides allows the support of these attributes. A main goal is to prepare a group of amide derivatives of carboxymethyl chitosan and carboxymethyl β-glucan to obtain a new group of polysaccharide derivatives and increase the benefits and attributes of these polysaccharides. The aim of carboxymethylation and subsequent two-step amidation is to achieve a high degree of substitution in the prepared derivatives. The amidation consisted methyl esterification followed by amino-de-alkoxylation with amidation reagents (n-alkylamines, hydrazine and hydroxylamine). The purity and substitution degree of the prepared derivatives were monitored by vibration spectroscopic methods (FTIR and FT Raman) and organic elemental analysis. The main, which was due to the presence of a secondary component (chitin) of polysaccharides, was the calculation of the substitution degree, which was based on the organic elemental analysis in combination with FTIR spectroscopy. These analytic methods confirmed the preparation of the substituted N-alkylamides, hydrazide and hydroxamic acid of carboxymethyl chitosan and carboxymethyl β-glucan.
- MeSH
- beta-glukany chemie MeSH
- chitin chemie MeSH
- chitosan analogy a deriváty chemie MeSH
- esterifikace MeSH
- hydraziny chemie MeSH
- hydrofobní a hydrofilní interakce MeSH
- lidé MeSH
- polysacharidy chemie MeSH
- spektroskopie infračervená s Fourierovou transformací metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy 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.
- 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
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.
- 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
Low molecular weight mannogalactofucans (LMMGFs) prepared by enzymatic degradation of high molecular weight Undaria galactofucan (MF) were evaluated for their anti-cancer effects against human prostate cancer. Correlation NMR and linkage analyses confirmed that LMMGFs consist mainly of α-fucose and β-galactose units: α-fucose units are 1,3-linked; β-galactose units are terminal, 1,3- and/or 1,6-linked; both sugars are partially sulphated, fucose at positions O-2 and/or O-4 and galactose at O-3. Mannose residue, as a minor sugar, presents as the 1,4-linked terminal units. LMMGFs more significantly induced cell cycle arrest at the G0/G1 phase and cell death via suppression of the Akt/GSK-3β/β-catenin pathway than MF in human PC-3 prostate cancer cells. LMMGFs upregulated mRNA expression of death receptor-5 (DR-5), the ratio of Bax to Bcl-2, the cleavage of caspases and PARP, the depolarisation of mitochondrial membrane potential, and ROS generation. LMMGFs (200-400 mg/kg) effectively reduced both tumour volume and size in a xenografted mouse model. These results demonstrated that LMMGFs attenuate the growth of human prostate cancer cells both in vitro and in vivo, suggesting that LMMGFs can be used as a potent functional ingredient in health-beneficial foods or as a therapeutic agent to prevent or treat androgen-independent human prostate cancer. Graphical Abstract.
- MeSH
- apoptóza účinky léků MeSH
- beta-katenin metabolismus MeSH
- kinasa glykogensynthasy 3beta metabolismus MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- messenger RNA genetika MeSH
- molekulová hmotnost MeSH
- myši MeSH
- nádory prostaty farmakoterapie metabolismus MeSH
- polysacharidy chemie farmakologie terapeutické užití MeSH
- Undaria chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
