Study On The Extraction Purification And Antitumor Activity Of Polysaccharides From Tricholoma Matsutake Fruitbodies

The fruit body of Tricholoma Matsutake is one of the most important edible fungis, which is not only very delicious and nutrimental, but also rich in polysaccharides that have multiple pharmacological activities and bioactivities. The activities of Tricholoma Matsutake polysaccharides (TMP) involve antitumor, enhancing immunity, antiradiancy and so on. In this study, the extraction, purification, modification, and antitumor activity of Tricholoma Matsutake fruit bodies were investigated. The main results are as follows:The optimal operation parameters for extracting were: the ratio of material to water is 1:30, the extraction temperature is 90℃, and duration is 3 hours respectively. Extraction twice is economy (the rate of the extractions is up to 94.3%). Precipitated the TMP with 3 times volume of 95% ethanol for 6h is the best choice in this study. The protein in the polysaccharides removed with the Sevag method. Ion exchange resin method was the best one in all of active carbon, H2O2 and the resin for decoloration by comparing the color and polysaccharides'loss rates. After the dialysis with the dialysis bag whose Molecular weight (MW) is over 8000, cellulose DE-52 column chromatography which eluted with of water and NaCl solution was used to separate fine polysaccharides. TMWEP-1,TMWEP-2,TMWEP-3 were obtained when the concentration of NaCl solution were 0, 0.2mol/L and 0.5mol/L, respectively. The TMWEP-2 was purified to TMWEP-2Ⅰand TMWEP-2Ⅱby Sephadex G-100 gel column. The TMWEP-2Ⅰwas identified as a homogeneous component by methods of Sephadex G-100 and HPLC. It was white solid after dialyzing, concentrating, and desiccating. TMP was modified with snail enzymes. The effect of modification mainly comes from four factors: duration, ratio of enzyme to TMP, pH and temperature. Via the mono-factor test and orthogonal test, we concluded that the optimum condition of modification were the duration=5h, the ratio=1:5, the pH=6.0 and the temperature=40℃. SGC-7901,Hela,HT-29 were three kinds of tumor cells. They were cultured and infused polysaccharides whose concentration were 10μg/mL, 100μg/mL and 1000μg/mL, and then identified the antitumor activities of TMP according to inhibitory rate to these tumor cells. Finally, the results showed that TMP has antitumor activity, and the inhibitory rate depends on the concentration and the purity; the degraded TMP by Snail enzymes has no activity. This study has done some basic work for the futher study and application of TMP.