Study On Extraction, Isolation And Purification Of Polysaccharide From Tricholoma Matsutake And Its Antioxidant Activity

Tricholoma matsutake,because of its delicious taste,tender meat,is popular with people.It is a kind of rare and precious natural medicinal fungi,known as "the king of edible fungi”.It contains polysaccharide,vitamin,amino acid,protein,anticancer and so on.It plays an important role in reducing blood fat,cardiovascular disease treatment,promoting gastrointestinal function.T.matsutake polysaccharide is one of the main active ingredients;it has high nutritional value and medicinal activity.T.matsutake has a broad development value and application prospect in health products,medicine and other fields.In this paper,we design five kinds of culture medium formula of T.matsutake mycelium in solid culture,and select the most suitable for the growth of T.matsutake solid medium culture,providing the basis for artificial cultivation of T.matsutake.We select the best extraction method by comparing the yield and antioxidant activity of T.matsutake polysaccharide in hot water extraction,ultrasonic aided the traditional hot water extraction,enzymatic extraction,microwave extraction method four kinds of extraction methods.The best extraction method is ultrasonic aided the traditional hot water extraction.Then we select the best extraction technology of ultrasonic aided the traditional hot water extraction by the single factor test and orthogonal test design.The method of alcohol classification is adopted in T.matsutake polysaccharide separation and purification,and we determine the composition of monosaccharide composition and antioxidant activity of T.matsutake polysaccharide.Maybe we can get the effective ingredients for the development of biomedical,providing a basis for further study about T.matsutake polysaccharide.The results of the study are as follows: 1.T.matsutake mycelium is cultivated under strict sterile conditions in five formula.According to mycelium growth(hypha density,morphology,hypha color,edge uniformity)and growth rate,we can know the growing condition of Tricholoma matsutake mycelium.Then we can select the most optimal mother kind of culture medium.It is PDA + peptone + malt juice + pine needles juice.2.The order of DPPH free radical removing ability and yield of T.matsutake polysaccharide is both for H-TMP > U-TMP > E-TMP > M-TMP.The order of total reducing power is U-TMP > H-TMP > M-TMP > E-TMP.The order of hydroxyl free radicals removing ability is U-TMP > H-TMP > E-TMP > M-TMP.Comparing with the yield and antioxidant activity of T.matsutake polysaccharide,we know that ultrasonic extraction yield and activity are relatively high.It can also save time and improves the efficiency than the traditional hot water extraction.3.An ultrasonic-assisted technique was employed to extract crude polysaccharide(TMP)from T.matsutake fruiting bodies.Single-factor tests and orthogonal experimental design(L9(3)3)were used to obtain the optimal extraction conditions.Results showed that the optimal parameters were as follows: ultrasonic temperature,40℃;ultrasonic time,50 min;water to raw material ratio,25 m L/g;ultrasonic frequency,45 k Hz;and ultrasonic power,100 W.Moreover,through verified tests,the highest extraction yield of polysaccharide TMP was 8.06%.4.Three novel polysaccharide fractions(TMP30,TMP60 and TMP80)were isolated and purified from TMP by stepwise alcohol precipitation.Their preliminary structural features were determined by HPAEC-PAD and FT-IR analysis.Furthermore,their in vitro antioxidant activities were investigated in terms of a reducing power assay and the scavenging rate of 2,2-diphenyl-1-picrylhydrazyl(DPPH)and hydroxyl radicals.The order of the various fractions based on their antioxidant activities was TMP80 > TMP > TMP60 > TMP30.These findings suggest that novel polysaccharide fractions from T.matsutake,especially TMP80,could be promising active macromolecules for biomedical use.