| Hericium caput-medusae is one of the valuable fungi in genus Hericium for both pharmacopoeia and edibility. Recently, polysaccharides of fungi in genus Hericium have attracted great attention owing to its effects on improving immunity, restraining cancer, reducing blood pressure and blood sugar, and so on. But nowadays, the research about Polysaccharides of Hericium caput-medusae are not enough, deeply studying on the extraction , isolation and purification combinding with biological activity will be important to production on a large scale. This article report extraction and isolation, purification, properties, some structural analysis and immunological action as adjuvant of Polysaccharides isolated from fruiting bodies of Hericium caput-medusae.Firstly,in order to remove lipid materials thoroughly,the fruiting bodies of Hericium caput-medusae were lixiviated four times by 95% ethanol. Then they were used as the marerial for experiment. We investigated the effec of extraction time, temperature, times , rate of material to liquid, precipitation with different ethanol on extraction rate. According to the results of single factors and the orthogonal design experiments, comprehensively taking all kinds of factors and conditions of production on a large-scale into account, we finally obtained the best technology for extraction of polysaccharide: The fruiting bodies of Hericium caput-medusae was extracted with 10 of water at 80℃ for 2 hours and repeated three times (the first time was 2 hours,the second time was 1 hour ,the third time was 1 hour), then concentrated to 1/5 of the former volume and precipitation with 70% ethanol. Production on a large-scale will benefit from this research.Through the best technology of extraction ,we obtained the crude polysaccharide(HPC ) by a yield of 8.2%. The extraction rate of polysaccharide of HPC was 5.7%.The polysaccharide and protein content of HPC were 70% and 4.4%,respectively. Compared with other crude polysaccharides of fungi in genus Hericium, the extraction rate of crude polysaccharide of HPC was lower.However, the purification was higher and protein content was less.The aqueous sugar solution was treated with Sevag reagent (polysaccharide liquid : Chloroform:n- butanol=5:1:0.2) for three times.The protein content amouted to 0.78% and were lower by 3.62% than before.Polysaccharides were fractionated by ethanol grading precititation,then we got HA (1% liquid of polysaccharide:95%ethonal =1:0.25) and HB(1%liquid of polysaccharide:95% ethonal =l:3).Through millipore filtration and ultrafication ,we got four fractions: HA-1 (ultrafiltrate that could not pass the membrane X HA-2(ultrafiltrate that could pass the membrane) and HB-1 (ultrafiltrate that could not pass the membrane) > HB-2(ultrafiltrate that could pass the membrane).Ultrafication was firsterly used in isolation and purification of polysaccharide of fungi in genus Hericium.The result showed that Ultrafication was a good method in isolation and purification of polysaccharide from fruiting bodies of Hericium caput-medusae and was adaptive for production on a large-scale.The identification of purity told us that HA^ HB-k HB-2 were uniform.The polysaccharide content of HA-2 was 98.6%. The estimated molecular weight of HA-2 was 50 690. SO42" content was 7.23%,optical rotation [a ]20D=—168.25. HA-2 was mainly composed of Man , Fuc> Glc > Gal, in the molar ratios of 1: 11.1: 10.8: 29.3.The main chain was made up of Fuc> Glc n Gal.The polysaccharide content of HB-1 was 99.8%. The estimated molecular weight of HB-1 was 100 000. SO42' content was 8.5%,optical rotation [a ]20D=181.82,. HB-1 was mainly composed of Man -. Fuk^ Glc ., Gal, in the molar ratios of 1: 4.3: 7.3: 13.7.The main chain was made up of Man ^ Fu^ Glc > Gal.The polysaccharide content of HB-2 was 99.6%. The estimated molecular weight of HB-2 was 30 200, optical rotation [a ]20D=-181.64, SO42" content was 5.37%. HB-2 was mainly composed of Man , Fucn Glc > Gal, in the molar ratios of 1: 5.8: 21.8: 20.3.The main chain was made up of Man .. Fuc> Glc -. Gal.The result of Infrared spectrum of HA-2^ HB-K HB-2 showed that they all had absorption at about 3400cm"1,2920 cm'1, 1580 cm"1, 1380 cm"1, 890 cm'1.So they had specific absorption of polysaccharide. Absorption at 1580 cm"1 showed they were protein-bound polysaccharides, probably. HA-2> HB-1 > HB-2 all exhibitated absorption at 890cm"1 , typical for P configuration.They had no a (1 -> 4) glucosidic bond.We used method of the sulfuric acid barium to determinate SO42" content of HA^ HB-1-. HB-2. The results were different.lt is important to investigate the biological activity of different component polysaccharides of different SO42" content.BALB/C mice were immunized with crude polysaccharide from fruiting bodies of Hericium caput-medusae(E?C) and BCG vaccine by hypodermic injection or mucous membrane route. In certain time, the weight, non-specific and specific responses were checked.The results showed that polysaccharides from fruiting bodies of Hericiumcaput-medusae could obviously improve non-specific and specific responses of BALB/C mice. At the same time, it had no obvious toxic function on BALB/C mice. Polysaccharide from fruiting bodies of Hericium caput-medusae can be used as an effective adjuvant to play more important role in the development of new-type vaccine. |
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