Study On The Fermentation Technology?chemical Constituents And Pharmacodynamics Of The Calvatia Lilacina

Objective:Isolation and screening of excellent strains of Calvatia lilacina.The biological characteristics of the screened strains of Calvatia lilacina were studied to master the nutrition and environmental conditions for the growth of its mycelium.The content of main chemical components in fermentation products to lay the foundation for its quality evaluation.The preventive and protective effects of Calvatia lilacina fermentation products on gastric ulcer and liver injury were studied,and to explore its mechanism of action,so as to lay a foundation for the development and application of its medicinal value basis.Methods:The excellent strains of Calvatia lilacina were isolated and purified,and their biological characteristics were investigated.The liquid fermentation process of Calvatia lilacina was optimized by single factor investigation and orthogonal experiment,and the composition and culture conditions of liquid broth were determined.The phenol-concentrated sulfuric acid method was used to determine the content of polysaccharides in the fermentation products of Calvatia lilacina.the color reaction of vanillin perchloric acid was used to determine the content of total triterpenes in the fermentation products of Calvatia lilacina.The content of ergosterone and ergosterol in fruiting bodies and fermentation products of Calvatia lilacina were determined by HPLC.The preventive effect and mechanism of the fermentation products of Calvatia lilacina on stress gastric ulcer and acute liver injury were studied through the stress induced gastric ulcer model induced by restraint water immersion and the acute liver injury model induced by alcohol in mice.Results:An excellent strain of Calvatia lilacina was obtained after isolation and purification.The biological characteristics of the strain showed that the most suitable medium for the growth of purple bald puff mycelia was glucose concentration of 2%,yeast powder concentration of 1%,and MgSO4 concentration of 0.1%,The initial pH value is 6.5,and the optimal culture temperature is 30°C.Studies on submerged fermentation of Calvatia lilacina showed that the optimum medium for liquid fermentation of purple puff was maltose 4%,yeast powder 3%,KH₂PO₄0.1%,initial pH value 6-7,optimum culture temperature 30 centigrade,and culture days 6 days.The content of polysaccharides in the fermentation broth was 17.71%and the content of polysaccharide in mycelia was 2.14%by phenol sulfuric acid method.The content of total triterpenoids in the mycelia of Calvatia lilacina was 2.50%by vanillin perchloric acid reaction.The content of ergosterol in mycelium is 4.97 mg/g,the content of ergosterone in mycelium is 11.86?g/g,the content of ergosterol in fruit bodies is 5.88 mg/g,and the content of ergosterone in fruit bodies 56.73?g/g.In the experiment of stress-induced gastric ulcer induced by water immersion in mice,the fermentation product of Calvatia lilacina reduced the levels of MDA and pepsin in gastric tissues,increased the levels of SOD and NO in gastric tissues,and reduced TNF-?and IL-6 levels,and down-regulate the expression of NLRP3,Caspase-1,ASC and IL-1?proteins.In experiments on alcohol-induced acute liver injury in mice,the fermentation product of Calvatia lilacina decreased the levels of AST and ALT in serum,reduced the levels of MDA,TB in liver tissues and the levels of inflammatory factors IL-18,IL-1?,TNF-?,and increased the level of SOD in liver tissues.Conclusion:The excellent strains of purple bald puppies were isolated and purified.The biological characteristics of the Calvatia lilacina were studied and the fermentation process was optimized to provide a basis for the subsequent industrial production of Calvatia lilacina.The content of each main component in the product provides a basis for its quality evaluation.In stress gastric ulcer experiments,the fermentation product of Calvatia lilacina can reduce the expression of NLRP3 protein and the adaptor protein ASC,inhibit the activation of the effector protein Caspase-1,and reduce the body's inflammatory factors IL-1?,TNF-?and IL-6 release,thereby effectively preventing the development of stress gastric ulcers in mice.In the experiment of acute liver injury,the fermented product of Calvatia lilacina may protect the mice with acute liver injury by regulating the levels of inflammatory factors and oxidative stress.