Study Of The Liquid Fermentation Of Tricholoma Matsutake And The Anti-tumor Activity Of Polysaccharides

Tricholoma matsutake is the most valuable mushroom exhibiting a characteristicand delicate flavor. The active ingredient polysaccharides have anti-tumor, antioxidantand improve the immune system and other active. In recent years, the fermentation ofmedicinal mushroom widely used to improve the fermentation conditions to improvethe quality of active ingredients, reduce fermentation costs, is the focus of the study ofliquid fermentation of the mushroom-borne fungi. This article will focus on the liquidfermentation and polysaccharide extract and anti-tumor activity studies of Tricholomamatsutake, the purpose is for it to even take full advantage of to provide a basis.On the basis of optimal carbon and nitrogen source, plackett-Burman design andBox-behnken design were applied to optimize the fermentation medium. Optimizedvalues of the factors were determined to be22.89g/L yeast extract,0.073g/Lcalcium chloride,0.053g/L potassium nitrate,2g/L magnesium sulfate,0.1g/L sodiumchloride,4g/L phosphatehydrogen potassium,25g/L lactose,0.15g/L of vitamin B1,and a maximum dry weight of mycelium of10.40g/L was obtained.On the basis of medium optimization, the shake-flask culture conditions wereinvestigated by means of single factor tests. The initial pH value in the culturemedium, seed age, inoculum size, medium volume, temperature, speed andfermentation time were all in consideration. The best culture conditionsas follows: pH6, temperature26℃, fermentation time of5days,7%of the speed of150r/min, andseed age5days, inoculum size, medium volume of100mL.Finally, Single factor tests and Box-behnken design were applied to optimize thehot water extraction and ultrasound extraction conditions of Tricholoma matsutakepolysaccharide.The best extraction conditions: ultrasonic time of202.43s, ultrasonicpower430.43W, water feed ratio of47.21:1, under the optimal conditions, the averageextraction rate reach13.186%. The hot water extraction conditions: extraction time4.49h, water and material than33.08:1, extraction temperature is91.95℃,, under theoptimal conditions, the average extraction rate reach12.84%. MTT assay was used tostudy the effect of Tricholoma matsutake polysaccharide on three types of tumor cells: human B16melanoma cells, human hepatoma cell SMMC7721, humancervical cancer cells Hela inhibitory rate. The results show that in vitro cell model, thepolysaccharide can significantly inhibit the proliferative effect of the three types oftumor cells, and the inhibition with the increase of polysaccharide concentration,showing a concentration-dependent. And when the polysaccharide concentrationreach10mg/mL, the inhibitory rate of the three kinds of tumor cells were63.54%,62.43%and57.81%, respectively.