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Sarang Cho,Peter Capek,Roman Bleha,Andrej Sinica,Jung Sik Lim,Jin Ree,Yong Il Park 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
Many plant polysaccharide have immunomodulatory and anti-tumour activities. In the present study, water-soluble polysaccharides were isolated from an edible plant seed mainly by hot water extraction, 75% ethanol precipitation, and DEAE-cellulose column chromatography and tentatively named as PSP-F1. The PSP-F1 was shown to be a mixture of three major polysaccharides with molecular mass of approximately 1,200, 640, and 300 kDa, respectively, as estimated by size-exclusion HPLC analysis. The HPAEC-PAD analysis for neutral sugar composition and showed that PSP-F1 consists of arabinose (Ara) and xylose (Xyl), as the major neutral sugars, and other minor neutral sugars such as rhamnose (Rha), galactose (Gal), and glucose (Glc). HPLC analysis for uronic acids showed that galacturonic acid (GalA) presents as the major uronic acid and glucuronic acid (GlcA) was the minor one. Together with the neutral monosaccharides and uronic acids composition, the FT-IR and NMR analysis suggested that PSP-F1 appears to be a mixture of pectin type and heteroxylan polysaccharides. The results showed that the PSP-F1 (0-100 μg/ml) significantly induced the secretion of pro-inflammatory cytokines TNF-α and IL-6 and upregulation of MAPKs (ERK, JNK and p38) phosphorylation. The secretion of various pro-inflammatory cytokines from macrophages is mainly caused by MAPK cascade activation. These results demonstrated that the PSP-F1 isolated from a plant seed is a mixture of pectin type and heteroxylan polysaccharides, which are typical for plant cell wall polysaccharides, and that it effectively stimulates macrophages to secrete pro-inflammatory cytokines, such as TNF-α and IL-6, by upregulation of MAPKs phosphorylation. The results of this study suggests that PSP-F1 may be potential immunostimulator for health benefits and as a pharmaceutical component.
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Isolation and characterization of bioactive polysaccharides from algae and medicinal mushrooms
Andriy Synytsya,GordonK.Gomba,Radek Pohl,Peter Capek,Jana Čopíková,Yong-Il Park 한국당과학회 2011 한국당과학회 학술대회 Vol.2011 No.1
Algae and mushrooms are rich sources of bioactive natural components. Among these compounds, polysaccharides are interesting due to their immunomodulating, antitumor, antiviral, coagulating and other properties. Polysaccharides play an outstanding role in the construction of algal and mushroom cell walls. First step of the strategy of polysaccharide isolation from such raw material is fractionation of individual cell wall components by subsequent extractions. The second step is rigorous purification of obtained fractions by using of preparative chromatography, chemical reagents and specific enzymes. Finally, individual polysaccharides within a fraction can be separated from each other by procedures based on the difference in their solubility in various media, sensitivity to chemical or enzymatic destruction, affinity to specific molecules and complexation with metal cations. Structural analysis of isolated algal or mushroom polysaccharides is based on combination of separation, chemical and spectroscopic methods. Total hydrolysis followed by HPLC or GC analysis gives the information about the monosaccharide composition. Gel chromatography equipped with various detectors is used for the molecular mass estimation. FTIR spectroscopy is effective for detection of functional groups, estimation of fraction purity as well as type of polysaccharide. Correlation NMR spectroscopy together with sugar linkage analysis is a powerful tool for detail analysis of polysaccharide structure (types of linkage and substitution, branching, anomeric configuration etc.). Polysaccharide fractions isolated from fruiting bodies of medicinal mushrooms Pleurotus ostreatus, Pleurotus eryngii, Agaricus blazei and Piptoporus betulinus were defined mainly as specific glucans. Branched 1,3-1,6-β-glucan and amylose-like 1,4-α-glucan were found in water soluble fractions, while linear 1,3-α-glucan in alkali soluble fractions. Effective deproteinisation and separation of α- and β-glucans was achieved by the treatment with phenolic reagent. Insoluble fractions contained some amount of chitin as a component of cell wall chitin–glucan complex. Fruiting bodies of wood decay fungi of genera Phellinus and Inonotus contains polyphenols integrated with cell wall polysaccharides. Most of polyphenols, which are interesting due to their strong antioxidative activity, were successfully removed from raw material by extraction with acidic ethanol. Soluble and insoluble polysaccharide fractions isolated from these mushrooms still contained colored products of polyphenol degradation. Polysaccharides were purified by the treatment with acidic or alkali solutions of hydrogen peroxide and defined mainly as branched β-glucans (soluble fractions) and chitin–glucan complex (insoluble part). Korean seaweeds Undaria pinattifida and Capsosiphon fulvescens are sources of bioactive polysaccharides, which were isolated and defined respectively as sulfated/acetylated galactofucan (fucoidan) and sulfated glucuronorhamnoxylan. These polysaccharides have shown various biological activities which make them interesting compounds for medicinal applications. This work was supported by Czech Science Foundation (Project No. 525050273), Ministry of Education of the Czech Republic (Projects No. CEZ: MSM6046137305) and Gyeonggi-do province, Korea (GRRC program of the Catholic University of Korea)
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Debjani Ghosh,Shruti S. Bandyopadhyay,Udipta R Chatterjee,Bimalendu Ray,Peter Capek 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.2
Chirata (Swertia chirata) is widely used in Indian folk medicine for the prevention of various disorders. Herein, we analyzed the water-extracted carbohydrate polymer (WECP) of this herb using chemical, chromatographic,and spectroscopic methods. The anti-oxidant capacity of this fraction was studied by ferric reducing anti-oxidant power (FRAP) and DPPH radical assays. Effect of WECP on bovine serum albumin spectrum (BSA) was determined using excitation wavelength of 282 nm. Anion exchange chromatography of WECP yielded 5 fractions (F1-F5) with different chemical compositions. The major fraction (F5)was homogeneous, had an apparent molecular mass of 4.5kDa, and contains both carbohydrates (57%) and phenolics (34%). The anti-oxidant capacities of WECP and F5 were comparable to standard anti-oxidants. Notably, activities of the carbohydrate polymers (F1-F5) correlate with their phenolics content. Fluorescence quenching measurement suggests that F5 can form complex with BSA and the value of the binding constant is K=6.28×105/M.
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