Study On Extraction, Purification, Classification And Bioklogical Activity Of Lentinan

Lentinula edodes,is the second most commonly cultivated edible mushrooms in the world. With high nutritive value and medical health care value, it is quite popular with the customers.Lentinan(LNT) is a higher molecular hyskon extrected from fruiting body and mycelium of Lentinula edodes. It exhibits prominent anticancer, antioxidant, regulating immunologic function activities, etc.This research adopted hot water extraction to optimize the extraction processes of LNT.Exploiting the orthogonal experimental design to definite the optimal parameter:extract temprature was 60℃,ratio of material and water(g/mL) was 1:10,extract time was lh,extract percent was 16.3%;The primary purification of LNT were carried out by DEAE-cellulose 52 chromatography,and collecting the effluent liquid from the same peak position, LNT-1 was gotten after concentrating and precipitating; LNT was modified by sulfating,then selected different molecular weights(MW) cut-off ultrafiltration membrane to classify it, six components with different MW were obtained; The single and symmetrical peak was gotten after detecting both LNT-1 and sulfated classified polysaccharides through Sephadex G-100 gelcolumn, which indicated the high purity;Scanning analyzing different products after extracting and purifying with Fourier infrared spectrum, all the absorption peaks possessed the characteristic of polysacchrides, which proved that they were polysacchrides;No typical absorption of nucleic acid or dissociated protein was observed in UV spectrum, so there was no nucleic acid and freeprotein;The experiment of anti-tumor activity in vitro of polysaccharides we found,in a concentration range of 100-1600μg/mL, LNT-1 and sulfation classified polysaccharides could both inhibit the growth of tumor cells;in 100～400μg/mL,the inhibition of the sulfation classified polysaccharide was not obvious, the maximal inhibition rate was 56.32%;in 800-1600μg/mL,the six components all showed significant inhibition, and the maximal inhibition rate was 85.05%;The maximal inhibition rate of LNT-1 to tumor cells was 83.98%,equaling to the components 3KD and 3-5KD,but higher than the other components. The results showed that LNT-1 and sulfation classified polysaccharides had certain inhibition effect on the growth of tumor cells in this concentration range,and and they possessed a positive correlation;In polysaccharide antioxidation testing, it was found that LNT, LNT-1and classified sulfated polysaccharides all had significant antioxidant activity, and the total antioxidant ability and reducing capacity were strong, while the inhibition ratio to hydroxyl radical and superoxide anion was low. The antioxidant activity of LNT,LNT-1 was stronger than classified sulfated polysaccharides.The results showed that the activity of LNT,LNT-1 and classified sulfated polysaccharides presented dose-effect relationship in certain concentration range.