Extraction,Purification And Biological Activity Of Triterpenoids From Morchella Mycelia

Morchella is one of the most popular wild fungi with both medicinal and medicinal properties.At present,many people have more and more knowledge about the health effects of their consumption.For example: The polysaccharides in Morchella esculenta have many effects on humans,such as improving immunity,alleviating fatigue,fighting cancer,inhibiting bacteria,and resisting oxidation.Physical health benefits.The Morchella contains a wide variety of beneficial ingredients for the human body.Experiments have shown that the content of the basic nutrient in fruiting bodies and fermentation cultured mycelium is almost the same between the cultivated Morchella esculenta fruiting material and the natural growing Morel.,Morel fruit bodies and mycelium are rich in nourishing and medicinal value of active substances.In this study,the wild Morchella fruiting body was used for tissue separation and spore separation,isolated and purified,and finally a Morchella strain was obtained.Among them,the effect of culture after separation from the cap was good.The strains obtained by the spore separation method showed milky white and dense growth.The isolated strains were then purified and the mycelium isolated from the spores grew densely and vigorously.The mycelium was cultured in a liquid medium.The morphology of the mycelial spheres obtained was the same as that of the standard Morchella mycelium spheres,indicating that the purity of the strains isolated from the spores was reliable.By morphological observation and microscopic observation,the morphological characteristics of the isolated mycelium and the standard Morchella strain were identical,indicating that the isolated strain was a Morchella strain.In this study,response surface methodology was used to optimize the fermentation conditions of Morchella mycelia,and the test and analysis were performed using Design-Expert 8.0 software.The optimum fermentation components predicted by the model are: soluble carbon source,beef extract,pH 6,shaker speed 140 rpm/min.The dry weight of Morchella mycelium was 3.82g/200 m L under the predicted conditions.The dry weight of Mycelium was 3.77g/200 mL,which proved the feasibility of the model.In this study,response surface methodology was used to optimize the extraction process of triterpenoids from Morchella mycelia,using Design-Expert 8.0 for testing and analysis.The optimal conditions for the model prediction were: ethanol volume fraction 60.90%,ultrasonic time 25.46 min,ultrasonic power 700.21 W,liquid-solid ratio 23.90 mL/g.Under this condition,the extraction rate of triterpenoids was 35.16 mg/m L.The verification test showed that the extraction rate of triterpenoids was 35.22 mg/mL,indicating that the model was effective.In this experiment,D-101 macroporous adsorption resin was used to purify the triterpenoids in Mycelium fermentation mycelia with good results.The eluent was selected for ethanol,and the concentrations were selected to be 30%,40%,50%,60%,75%,and 95%,respectively.Gradient elution was performed on the total crude triterpenoids,of which 60%,70%,and 95% ethanol were all available.Triterpenoids are eluted.The DPPH method was used to quantify the anti-oxidation activity of triterpenoids in the effluent.The clearance rate was close to 40%,indicating that the eluted fractions all had higher antioxidant activity.The results of high performance liquid chromatography(HPLC)analysis showed that the triterpenoids contained in the collection solution of the 87-tube and the 123-tube collection were betulinic acid.In this experiment,the triterpenoids extracted from Morchella mycelia were tested for their antioxidant activity and biological activity of anti-tumor cells.Among them,DPPH free radicals,total reducing power,and super-activity were used to determine the antioxidant activity.Oxy-anion radicals,hydroxyl radicals,as a whole,triterpenoids extracted from Morchella mycelium have certain antioxidant activity.At the same time,it can also significantly inhibit the proliferation of HT-29 cells,HepG-2 cells,PC-3 cells and HeLa cells.The IC50 of the four tumor cells were 1.79,3.06,1.31 and 4.23,respectively,and they inhibited PC-3 cells.The effect is best.