Studies On Preparation Of Sulfated Polysaccharides And Activity Of Hericium Caput-medusae (Bull.:Fr)Pers.

Hericium caput-medusae is one of the valuable fungi for both medical and edibility, It is scarce fungus natural resources in Mount Changbai natural reserve. Hericium caput-medusae include many kinds of nutritive material, of which polysaccharide component is major studied, its activity has many aspects about increasing immunity,resisting cancer,anti-oxidant,enhancing digestive function and so on. Nowadays, the research about extraction method,structures research,bioactivity aspects, but studying on chemical modification of Hericium caput-medusae on a large scale. This study used two methods to preparation sulfated derivatives, then the biological activities of Hericium caput-medusae and sulfated derivatives were determined. It will impulse the development of Hericium caput-medusae polysaccharide about the aspect of pharmacology and biologic. On development medical value of Hericium caput-medusae to provide accordance with theory.Warm water extraction was used for hericium caput-medusae liquid submerged fermentation extractum polysaccharides, based on solvent-solid ratio,extracting time,extracting temperature single-factor experiments, The SAS 9.2 software was employed to design response surface, the yield of polysaccharides was used as the responsive values, The optimum extraction conditions by optimize were as follow: solvent-solid ratio 2.29:1 (mL/g), extracting time 4.52 h, extracting temperature 79.5℃, the yield of polysaccharides rate was 12.30%. Through ethyl alcohol fractional precipitation, to choose 75% fraction further purification on the base of yield, removing floating protein by Sevag method. Dialysis select bag filter which is MW 8000-10000. Selection Sepharose CL-6B gel column chromatography was used to fractionation on polysaccharides, collecting the main peak, The result of identification the purity of polysaccharide is uniform polysaccharide, named HLSF. Gel chromatography is used to measure the molecular weight of polysaccharide is 4.32×104Da.HLSF was sulfated using Wolfrom method and sulfuric acid method, to obtain HLSFL and HLSFN respectively. The result is analyses by infrared spectrum and substitution, it has the special absorption peak about sulfate, to prove the sulphuric acid group has linked with the polysaccharide, the products has named HLSFL and HLSFN respectively. Through the MTT experiment in vitro and SOD kit analysis comparing the activity of polysaccharide sample which is modified or not, the result of MTT experiment in vitro displayed the polysaccharide sample which is sulfated has more activity to the cell inhibition ratio of MCF-7 than the polysaccharide sample. When the polysaccharide concentration is 1200μg/mL, the inhibition ratio of HLSFL is improving to 27.00% to the cell of MCF-7, the inhibition ratio of HLSFN is improving to 21.21% to the cell of MCF-7, The result of sample polysaccharides inhibition ratio of the activity to the cell of MCF-7 by sulfating modification is enhanced obviously. SOD kit analysis comparing the activity of polysaccharide sample to remove superoxide anion which is modified or not. The activity of HLSFL is improved 102.49% than sample polysaccharide, The activity of HLSFN is improved 27.53% than sample polysaccharide, the result indicated the activity of anti-oxidant of sample polysaccharide by sulfating modification is enhanced obviously. Furthermore the activity of the sulfating derivatives by Wolfrom method is more higher than the activity of the sulfating derivatives by sulfuric acid method. As all the result, the bioactivity of sample polysaccharide sulfate is better, Wolfrom method is more fit to prepare polysaccharide sulfate of Hericium caput-medusae liquid submerged fermentation extractum than the sulfuric acid method.