Study On Extraction-Separation And Immune Activity Of Fermentative Cordyceps Polysaccharide

As a rare tonic Chinese herbal medicine,cordyceps sinensis has many kinds of biological activities,and cordyceps polysaccharide is one of the major components of these therapeutic effects.Natural caterpillar fungus due to limited by conditions can not meet the rapidly growing market demand.Instead of Chinese caterpillar fungus,fermentative cordycepic fungal powder has a great application prospect.At present,which scope of molecular weight polysaccharides play the effect of immunization is not clear.Taking fermentative cordycepic fungal powder as raw material,using the orthogonal method to optimize the microwave extraction technology of cordyceps polysaccharide,coupled with ultrafiltration technology for the isolation and purification,and using infrared technology analysis the structures of graded polysaccharide,finally,according to results of the anti-stress effect,allergy,humoral immunity and cellular immunity,etc,screening immune activity of hierarchical polysaccharides.Using fermentative cordycepic fungal powder(Cs-4)as raw materials,single factor experiments and orthogonal method be used to optimize microwave extractive process conditions,and combined with the actual industrial production situation,it concluded that the best technologies of cordyceps sinensis polysaccharide by microwave assisted extraction are solid-liquid ratiol:10(g/mL),microwave power 400W,pH=9,microwave time five minutes.In this conditions,the average of the polysaccharide content with three parallel experiment results is 53.149%,higher than the results of other single factor tests.Using the best technological parameters to extract polysaccharide largely,with simple preprocessing to remove impurities such as proteins,and different range of molecular weight cut off for ultrafiltration membrane be used to separation fermentative cordyceps polysaccharide.Different molecular weight ranges of FCP dry matter accounted for the proportion of total polysaccharides as follow:the molecular weight of 100kD or accounted for 9.03%,FCP60?100kD for 20.73%,FCP20?60kD for 24.498%,the molecular weight of 20kD or less for 45.739%,and intercepted mass fractions of the dry matter polysaccharides were above 85%.Infrared technique find graded characteristics components have the characteristics of the saccharides functional groups.Study on total polysaccharide and four graded characteristics for atmospheric pressure hypoxic tolerance in mice anti-stress activity screening experiment and a series of immune efficacy experiments.Testing methods including carbon clearance method,evans blue colorimetry and H-E staining method and so on.Experimental results show that all of the polysaccharide components have certain anti-stress effect;Molecular weight less than 60kD of polysaccharide components can significantly improve the index of phagocytosis in mice,and can weaken the delayed-type skin allergic reaction induced by DNCB.HE staining results show that the immuno-suppression model was established successfully with hydrocortisone stimulation mice;Total polysaccharides and FCP20kD all have good immune effects,but is inferior to positive group.FCP20?60kD effect is slightly better than the normal control group.And the effects of rest groups are not obvious to normal group.