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Structural analysis of polysaccharides from mushroom hericium Erinaceus mftccb 134
Bleha, R. ; Čopíková, J. ; Sushytskyi, Leonid ; Vannucci, Luca ; Sinica, A. ; Jozífek, M. ; Jablonský, I. ; Klouček, P.
The aim of this research was the structural characterisation of polysaccharides isolated from fruiting bodies of the cultivating medicinal mushroom Hericium erinaceus MFTCCB 134. Cold and hot water extracts (CWE, HWE) were obtained and purified, and followed by alkali extraction from the reminder solids. The composition and structure of the fractions were analysed by FTIR spectroscopy, and neutral sugars/linkage analyses were determined by GC/FID and GC/MS, respectively. Galactose, fucose and glucose were found in purified CWE,\nwhile glucose predominated in purified HWE.
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Purification of water soluble bioactive polysaccharides from biomass of heterotrophic mutant alga Chlorella vulgaris G11.
Sushytskyi, L. ; Lukáč, Pavol ; Bleha, R. ; Čopíková, J. ; Vannucci, Luca ; Sinica, A. ; Kaštánek, P.
Water extract obtained from biomass of mutant alga Chlorella vulgaris G11 was deproteinized and then purified with AEC on DEAE Sepharose FF and SEC on Sepaphadex G-75 superfine columns. Two high MW (>1000 kDa and 98,3 kDa) polysaccharides (CPS1 and CPS3) were eluted with water and 0.5M NaCl on DEAE column, and heteropolysaccharide with MW of 15 kDa (CPS2) was also eluted with water on DEAE. The former fractions were identified as branched rhamnans, the latter one as branched xylogalactofuranan of unusual structure. CPS1 shown properties of unstable polimer: according to GPC analysis and by using light scattering and RI detectors it was found, that some fraction of polymers (260-800 kDa) have self-assembling properties in aqueous solution, and MW of this fraction decreased during some time while polimer with initially bigger MW (>1000 kDa) became bigger. CPS3 was eluted with 0.3-0.5 M of NaCl on DEAE Sepharose FF column and have MW of 98,3 kDa. Cell toxicity of the products were tested in vitro on cell cultures for different concentrations
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Structure of an immunoactive polysaccharide isolated from Korean mulberry fruit (Morus alba L.)
Bleha, R. ; Lee, J. S. ; Capek, P. ; Pohl, Radek ; Kim, H. B. ; Choi, D. J. ; Lee, S. ; Lee, J. ; Jang, S. J. ; Synytsya, A. ; Park, Y. I.
A water-soluble polysaccharide (JS-MP-1) was isolated from the water extract of Korean white mulberry fruits (Morus alba L.) by ethanol precipitation and then purified by DEAE-cellulose ion exchange chromatography. Obtained final polysaccharide (1600 KDa) consisted of galactose, arabinose and rhamnose as major neutral sugars. JS-MP-1 also contains galacturonic and glucuronic acids (4:1). Sugar linkage, FTIR and correlation NMR analyses confirmed that it is a rhamnogalacturonan type I (RG I), which contains the alternating sequence of 1,4-alpha-D-GalAp and 1,2-alpha-L-Rhap units in the backbone. Neutral sugar side chains of JS-MP1 were identified as (1 -> 5)-alpha-L-arabinan and arabinogalactan type II (AG II) having the (1 -> 6)-beta-D-galactan core. The arabinan side chains are bound to the backbone at the O-4 position of some alpha-L-Rhap units, while the way of linkage between RG I and AG II chains is unclear. It was demonstrated that JS-MP-1 significantly stimulates murine macrophage RAW 264.7 cells to release chemokines (RANTES and MIP-1 alpha) and proinflammatory cytokines like INF-alpha and IL-6, and induce the iNOS and COX-2 gene expression, which are responsible for the production of NO and prostaglandin PGE2, respectively. These results suggest that JS-MP-1 can act as a potent immunomodulator and these observations may support the applicability of this polysaccharide or the water extracts of mulberry fruit can be used as an immunotherapeutic adjuvant or health beneficial food material.
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