Extraction,Purification,Structure And Bioactivities Of The Polysaccharides From Ganoderma Lucidum Mycelium

Ganoderma Lucidum is a traditional precious herb with a variety of biological activities which has been widely recognized by researchers.Ganoderma Lucidum polysaccharides as the major bioactive components of Ganoderma Lucidum attracted tremendous attention.In this study,the extraction,isolation and purification,physical,chemical properties,structure characterizations,immune and anti-hemolytic activities of the polysaccharides from Ganoderma Lucidum Mycelium were investigated.The results showed that:On the basis of single factor,the L9(34)orthogonal test was used to determine the extraction yield of Ganoderma Lucidum Mycelium polysaccharide.The results showed that the solid-liquid ratio was 1:30(W/V,g/ml),the time was 4 h,the temperature was 90 ?,the pH was 7.0 and the yield reached to 9.56%.The results of qualitative analysis,ultraviolet spectroscop,and congo red test showed that there was no starch,phenol,nucleic acid,protein or three-dimensional spiral structure in Ganoderma Lucidum Mycelium polysaccharides,while existence of specific polysaccharide structure was noticed.The SEM results showed that the surface structures of LZa-1,LZb-1and LZc-1.LZa-1 was all coral shape and smooth with some small vesicles on it,and no pore openings was observed.LZb-1 was disc shape,its surface was smooth with some small vesicles,some had concave and hollow on it.LZc-1 was cypress leaf shaped with some rod-like structures and some small elliptical shapess on it and the end of which is round and unbranched.Infrared spectrum analysis showed that all the Ganoderma Lucidum polysaccharides presented the characteristic absorption peak of polysaccharide structure,including ?-pyranoside in LZa-1,pyran ring and mannose in LZb-1 and ?-pyranoside,?-pyranoside and mannose in LZc-1.LZa-1,LZb-1and LZc-1 were determinated by GC-MS and the results showed that those 3 Ganoderma Lucidum polysaccharides mainly consisted of rhamnose,fucose,mannose,glucose,galactose and arabinose and the molar ratio of monosaccharide of LZa-1,LZb-1 and LZc-1 were 0.94 : 0.50 : 1.68 : 26.91 : 4.80 : 17.12,1.05 : 1.56 : 20.66 : 1.97 : 5.77 : 12.16,1.53 : 0.61 : 4.02: 5.16 : 4.25 : 23.28 respectively.The periodate oxidation and Smith degradation results showed that the main linkage types of LZa-1were proved to be 1?2,1?3,1?4 and 1?6.The main linkage types of LZb-1 were proved to be 1?2 or 1?6,1?3.The main linkage types of LZc-1 were proved to be 1?4,1?2 or 1?6,1?3.The 1H and 13 C NMR results showed that LZa-1,LZb-1 and LZc-1 appeared the chacteristic peak of sugar.The results of glycosidic bonds,monosaccharide compositions,anomeric carbons were consistent with those measured by IR,GC-MS and Smith degradation.Different structure of LZa-1,LZb-1and LZc-1 leaded to different biological activity.The RAW264.7 model showed that LZa-1,LZb-1 and LZc-1 had strong immunomodultory activity.They can improve the secretion of the NO,TNF-? and IL-6 and the effect was dose-related.LZa-1,LZb-1 and LZc-1 also showed protective effect against oxidative hemolysis induced by AAPH.They could effectively inhibit AAPH-induced overproduction of reactive oxygen speciesand alleviated the enhancement of intracellular antioxidant enzymes(SOD,GPX,CAT)activities.Meanwhile,the intracellular MDA formation were also inhibited by their treatment which contributed to the protection of the cell structure with the LZa-1 the most predominant polysaccharid.