| Carya Cathayensis Sarg.is an important economical plant in Zhejiang Province.It is delicious and nutritional.But its husk is thrown away as rubbish,which is pollution to the environment and waste of useful resources.So an efficient method is needed to explore and use the carya husk.With the development of glycol-biology,polysaccharides isolated from botanical sources are showing a broad spectrum of therapeutic properties and relatively low toxicity thus they have attracted a great deal of attention in the biomedical arena.Therefore development and utilization of carya polysaccharide has theoretical and practical significance.Polysaccharide was extracted and isolated from Carya cathayensis Sarg.,in this paper,its structural properties and bioactivities were analyzed further.According to the orthogonal experiment,the optimal extraction parameters were established,they are:ratio of water and materiel 1:15, extraction temperature 70℃and extraction time 150min.The extracted crude polysaccharide was named as PSW.In the deprotein study,TCA method and Sevag method has been compared,in consideration of reagent waste and the effect on structure and bioactivity of polysaccharide,it is determined to treat the sugar solution with Sevag solution 7 times as the optimal deprotein method. The further purify methods such as decolorization,dialysis and column chromatography separation was explored in this paper.Deproteined solution of sugar is decolored with 10%active carbon at 60℃for 2h, dialysis with dialysis bag of 14000 molecular weight,and then purified with DEAE sepharose FF ion exchange column(2.0×20cm)and Sepharose 4B gel colunm(2.0×40cm).The purified polysaccharide was determined to be a pure one according to the analysis result of HP GPC and CAME,it's named as PSW-1.IR spectrum indicates that PSW-1 is an acidic polysaccharide with uronic acid.The physicochemical analysis result shows that PSW-1 contains 61.9%uronic acid,and its C/O value is near 1,which means PSW-1 may be a heteropolysaccharide contains glycuronic acid.The neutral monosaccharide composition of PSW-1 is 23.0%glucose,44.4% mannose,20.0%galactose and 12.6%rhamnose in terms of the molar proportions according to the GC data.HP GPC analysis shows that the molecular weight of PSW-1 is 133 021D.The previous polysaccharide research showed that most of the hetero-polysaccharide has strong bioactivity while the polysaccharide with molecular weight between 100 000～200 000 D has the strongest activity,so based on the above experimental results,PSW-1 can be speculated with potential high bioactivity.In the antioxidant test PSW-1 showed antioxidant capability by scavenging 54.4%DPPH Free Radical at 1mg·mL-1,it may has the advantage of mildness and nontoxicity as a botanic source which can be used.PSW-1 showed stronger immuno-modulation effect than PSW in the immune test in vitro,it can produce statistically significant proliferation of murine spleen lymphocyte at both 10.00μg·mL-1and 100.00μg·mL-1 while PSW only at 100.00μg·mL-1.Both PSW-1 and PSW can significantly stimulate the growth of murine spleen lymphocyte induced by ConA or LPS at 0.01,0.10,1.00,10.00 and 100.00μg·mL-1,and the ConA-induced lymphocyte showed more significant proliferation after treated with PSW-1,which means PSW-1 aims more at T Cell.In Delayed Type Hypersensitivity(DTH)responses analysis,PSW-1 stimulates DTH response in mice at a dose of 80mg·kg-1·day-1,which means PSW-1 can efficiently produce T-Cell proliferation.
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