Isolation And Activity Study Of Secondary Metabolites From Volva Of Phallus Dongsun

Fungi are widely distributed in nature,the total number of species is estimated to be about 1.5million,known species account for only 5%of them,is a very large number of biological groups.In the past 50 years,most of the new drugs for clinical use come from natural products or natural product derivatives.The importance of fungi as producers of natural products is increasing,accounting for 61%of all bioactive compounds found since 2001,but compared with this large group,at present,the excavation of the chemical composition and biological activity of fungi is still very limited,so it is of great significance to study the active material basis and activity spectrum of fungi.The main research objects of this subject is volva of Phallus dongsun.Phallus dongsun also known as Phyllostachys pubescens,Dictyophora przewalskii,is a fungus belonging to Basidiomycotina,Gasteromycetidae,Phallales,Phallaceae and Phallus.It is a kind of large fungus.Previous studies have shown that Dongsun fungus receptacle contains a large number of active substances such as polysaccharides,flavonoids,phenols,terpenoids and sterols,and the extract has strong antioxidant activity,anti-tumor and anti-inflammatory activity.However,due to the poor taste and difficult to deal with,nearly 70%of the fungus and sporophyte were discarded after picking.This not only causes a waste of edible fungus resources,but also causes pollution of the planting environment.In order to explore how to make effective use of the fungus receptacle,this project plans to start with these active components.A total of 21 compounds(1-21)were isolated and identified from the ethyl acetate layer of the crude extract of Dongsun fungus,including 7steroids,4 terpenoids,2 alkaloids,2 carboxylic acids,3 fatty acids and 4 other compounds.One of the terpenoids was identified as a new compound by NMR,high resolution mass spectrometry and circular dichroism spectroscopy,and the rest were known compounds.The antibacterial activity,anti-inflammatory activity,antioxidant and anticancer activity of the isolated and identified compounds were evaluated.The antibacterial activity of the compounds isolated from the crude extract of ethyl acetate layer was determined.Five strains of human pathogenic bacteria,including Staphylococcus aureus(ATCC13565),MRSA(ATCC6538),Proteus Proteus(FY012),Streptococcus agalactis(ATCC13813)and Pseudomonas aeruginosa(ATCC27853),and a strain of trichoderma,the pathogen of Dictyophora rubrovalvata,were selected.The antibacterial activity was screened by Agar diffusion method,and the MIC of the compounds with inhibitory activity was determined by 96-well plate microdilution method.It was found that some compounds had certain inhibitory effects on Streptococcus agalactis,Staphylococcus aureus,Pseudomonas aeruginosa,MRSA and Trichoderma.The inhibitory effects of compound 18~*on leukemia HL-60,lung cancer A549,liver cancer SMMC-7721,breast cancer MDA-MB-231 and colon cancer SW480 were tested by MTS method.The results showed that at40μM,the new compound 18~*had no inhibitory effect on all anti-tumor cells in vitro,and some known compounds had certain inhibitory effects on hepatoma cells SMMC-7721,leukemia HL-60cells and colon cancer SW480 cells.The antioxidant activity of some compounds was studied by FRAP method.The results showed that the new compound 18~*had weak antioxidant activity and some compounds had moderate antioxidant activity under 15 mg/ml.The anti-inflammatory activity of the compound was indirectly demonstrated by detecting the inhibition rate of NO production of RAW264.7 in mouse mononuclear macrophages induced by LPS lipopolysaccharide.The results showed that the new compound 18~*had no anti-inflammatory activity at 50μM,and some known compounds had moderate NO inhibitory activity in vitro.In this paper,through the study on the chemical composition and biological activity of Phallus dongsun’s volva,the potential of antibacterial,antioxidant,anti-tumor and anti-inflammatory activity of waste fungus was preliminarily verified from the small molecular level.It provides clues and theoretical basis for the rational development and utilization of the waste fungus.The effective use of these active components can not only improve the income of bacterial farmers,but also is of great significance to the regeneration of ecological resources.