| Pholiota dinghuensis Bi is a kind of macrofungi. This is an special breed in China. It is near Qingyun temple which on Dinghu mountain in Guangdong province. The fungi of pholiota have a lot of medicinal values and healthy functions.At present there are a few reports of research on dinghu pholiota, this dissertation is about the research which on aspects as formula of Pholiota dinghuensis Bi’s medium, fermentation condition, separation purification and extraction of polysaccharides in mycelia as well as the biological activity of polysaccharides of Pholiota dinghuensis Bi, and the main content are as follows:1. Pholiota dinghuensis Bi’s screening of polysaccharide medium and optimization of the condition of fermentation.Firstly, compound potato dextrose angular surface medium (PDA medium); secondly, preparation liquid seeds medium, thirdly make single factor experiment with carbon source, nitrogen source, inorganic salts in the fermentation medium which with the method of three factors three levels orthogonal, at last we can draw the best medium formula:carbon source of glucose is40g/L; nitrogen source of peptone-yeast extract complex,both ratio is1:1, each for4g/L; inorganic salts are magnesium sulfate(MgSO4) and potassium dihydrogen phosphate(KH2PO4),the proportion were1.5g/L and1g/L. PH is original, fermentation temperature is28℃, incubation time is7day,rotation rate at150r/min.Through screening the best formula of fermentation medium, then do research with the Pholiota dinghuensis Bi’s fermentation conditions, which the different fermentation temperature, fermentation time, and stirring speed, the result showed that with training8day on temperature of28℃, inoculation level of15%, stirring speed of150r/min, aeration rate as0.75vvm, in this condition the maximum yield of fermentation mycelia is10.98g/L.2The extraction, separation and purification of polysaccharide in Pholiota ding huensis BiIt was found that ultrasonic extraction yield is bigger than the another and use less time when we separately use hot water extraction and ultrasonic extraction with the conditions of same temperature, time, and the ratio of the liquid and material.At first, selecting four factor which are extraction temperature, extraction time, the ratio of the liquid and material and ultrasonic extraction time as single factor experiment, then on this foundation, choose extraction temperature, extraction time and water material than do experiment with the method of L9(34) of three factors three levels orthogonal. And at last we draw that the optimal extraction conditions for polysaccharide extracting of Pholiota dinghuensis Bi is extraction temperature on90℃, extracting time for5hour, the ratio of the liquid and material is of20ml/g and ultrasonic extraction time is50minute.After using DEAE-cellulose-52and glucose G-100chromatographic column to PDP impurity products for preliminary separation and further purification, we get three groups which were PDP-1, PDP-2and PDP-3. The yield rate of PDP-1, PDP-2and PDP-3were60.23%,23.68%and4.63%.3In vitro antioxidant and in vivo immune activity of Pholiota dinghuensis BiThe results show that the pure polysaccharide PDP-1, PDP-2and PDP-3can remove hydrogen peroxide (H2O2), DPPH free radicals, chelate ferrous ion (Fe2+),Remove hydroxyl radicals and have certain of in vitro antioxidant effect when we use chemical method to measurement and analysis in vitro antioxidant activity of pure Pholiota dinghuensis Bi polysaccharide PDP-1, PDP-2and PDP-3.When we use mice model test to studying and analysis thick PDP in vivo immune activity, results show that PDP can recover the immunity which caused by production of cyclophosphamide induction,and at the same time it can dramatically improve the index of spleen, right ear swelling of the late-noset hypersensitivity, lysozyme in spleen and serum, and this kind of transformation relationship rely on dose of impurity Pholiota dinghuensis Bi polysaccharide. Impurity Pholiota dinghuensis Bi polysaccharide has certain of in vivo immune activity. |
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