Study On Purification And Characteristics Of Extracellular Polysaccharid In Tricholoma Matsutake

Tricholoma matsutake is a kind of fungus which can be used as food and medicine, people love it because of its rich nutrition,unique flavor and many physiological functions. As a living symbiotic bacteria, it is difficult to isolation and cultivation for Tricholoma matsutake strains. So artificial cultivation technique of Tricholoma matsutake have not been come ture at present,which is restricted the development and application of Tricholoma matsutake series products on certain levels. It is showed that, polysaccharide is the main active components of Tricholoma matsutake.Now we pay more attention on polysaccharide in Tricholoma matsutake mycelium and fruitbody, but less on extracellular polysaccharide. In this paper,we used the fermentation liquid as the main raw materials, researched on the extraction and purification of matsutake extracellular polysaccharides,and the physiochemical characteristics of purified polysaccharide. The researches are provided the theory basis for further purification,extraction and structure analysis to active substances of Tricholoma matsutake,layed a good foundation on the development and utilization of Tricholoma matsutake extracellular polysaccharide and series of functional food and drugs,.The main research results are as follows:1. The process conditions of Tricholoma matsutake extracellular polysaccharide extraction were optimized by orthogonal experiment,the best conditions are:final ethanol concentration60%, pH8, alcohol sedimentation time9h.Under this conditions,the extracted volum of Tricholoma matsutake extracellular polysaccharide can be researched to0.412g/L.2. Using Sevag method, activated charcoal,dialysis,DEAE-52cellulose ion exchange column chromatography and Sephadex G-100column chromatography to isolate and purifiy, TMS Ⅰ-A and TMS Ⅱ were obtained, which purity quotient is92.1%and90.4%respectively. TMS Ⅰ-A is white powder and TMS Ⅱ is light yellow powder, two of them are light, no special smell.These can be dissolved in water, easily solubled in hot water, insolubled in ethanol, acetone, ethyl ether, chloroform, ethyl acetate, dimethyl sulfoxide and so on. These are acidic polysaccharides which do not contain protein, nucleic acid,.3. The molecular weight determination for two kinds of polysaccharides are used gel filtration method, the results showed that:the molecular weight of TMS-A is68kD, the molecular weight of is32kD.4. The results of UV spectra showed that TMS Ⅰ-A and TMS Ⅱ have absorption peak, of polysaccharides. The results of infrared spectroscopy revealed that the purified polysaccharide of TMS-A and TMS Ⅱ are a type of dextrose and pyranoid with carbohydrate characteristic absorption peak.5. The analysis of high performance liquid chromatography on the composition in matsutake extracellular polysaccharide monosaccharide showed that, the matsutake extracellular polysaccharide of TMSI-A composed of glucose, mannose, xylose, which is41%,28.60%,15.99%respectively. TMS Ⅱ is also mainly composed of glucose, mannose, xylose, which is53.72%,27.92%,16.46%respectively.