Study On The Mutation Breeding, Liquid Fermdntation And Biological Activities Of Polysaccharide From Poria Cocos

To explore t he application of the ult raviolet ray induced mutation in breeding good st rain of Poria cocos, good st rain P6 has been obtained t hrough t he screening model of good st rain after sporesuspension of Poria cocos had been irradiated by ult raviolet ray. It s dry weight of filamentous formreached 11.92 gram per liter after st rain P6 Erlenmeyer flask liquid fermentation, which was increased by 7.58% compared wit h t he original st rain PO.After st rain P6 subcult ured, it was proved t hat good st rain P6 had good genetic stability by st rain P6 Erlenmeyer flask liquid fermentation.The six Poria cocos strains from different sources were rejuvenatived in test tubes equipped with PDA culture medium. According to the fermentation broth pH.reducing sugar content and biomass.the growth line of Poria was drew;Observe and record the influences of different spawn age of solid medium slope. Single factor tests were done to analyze the influence of each factor including temperature.rotation speed,inoculum size.medium volume.carbon source and nitrogen source.Then the optimum conditions for the fermentation were obtained by using orthogonal design L934. The results of single factor tests and orthogonal experiments demonstrated that the optimum fermentation conditions are as follows:temperature is 26℃.rotation speed is 140r/min.inoculum size is 10%.medium volume is 100ml/250ml; 2.5% glucose,2%extractum carnis,KH2PO4/MgSO4 is 0.6%/0.3%, VB1 is 4mg/100ml. Fermentation growth lineof Poria was drew;determine the optimum spawn age (25d) of solid medium slope.Carboxymethylpachymaran which had more biological activities was obtained by structure transformation of pachyman which was isolated by fermentation-alcohol-deeping from fermentation broth.Through multi-step purification.component analysis,determination for physical, chemical and structure characteristics.CMP was examined.The results indicated that CMP was pure polysaccharide composed of carboxymethyl glucose only.with no evidence showing the existence of nucleic acid or protein,PG wasβ-type glucosan with its glucosidic bond was connected byβ(1→3),PG was glucopyranose and its molecular weight was about 4.2×104 Da.optical rotation was-3.1°.the viscosity was 62.55.Compared with the original pachyman,there have been some changes in structure and characteristics of CMP.The polysaccharide carboxymethyl transformation had achieved certain result.Preliminary results showed that poria polysaccharide did not have clear antibacterial activity (.3) From MTT assay.PORIA COCOS can restrain HepG2 and breast carcinoma (MDA-MB-231).with 1C50 74.1 ug/ml and 152.4 ug/ml,respectively.PC2 can inhibit breast carcinoma also,IC50 is 897.4 ug/ml.PORIA COCOS has high inhibiting intensity in the first six hours.after adding PORIA COCOS to the cells in vitro.The estimated average molecular weight of PORIA COCOS is 1×104 by viscosity measuring method.Through dialysis we found that different components of PORIA COCOS in inhibiting breast carcinoma and HepG2,are differently.The result speculates that poria polysaccharides for different cancer their bioactivities may differ.and their specific mechanism may also be different.