Secondary Metabolites From Trichoderma Atroviride D16 And S361

Plant endophytic fungi are fungi that survive in healthy plant tissues at a certain stage of life history,which do not cause obvious symptoms or damage to their hosts.Salvia miltiorrhiza Bge.is a dicotyledons Labiatae family erect perennial herba ceous plant,a kind of common traditional Chinese medicinal plants,with many treatment effects such as blood stasis,dredging the pain,heart chufan,liangxue carbuncle.Cephalotaxus fortunei Hook.f.is an evergreen tree which is found in subtropical area,surviving in Zhejiang,southern Anhui,Fujian,Jiangxi,Hunan and other provinces.It possesses the activities of insecticide,xiaoji,anti-cancer,used for the treatment of cough due to indigestion,roundworm,hookworm disease.Secondary metabolites are compounds with complex structures,produced by some microbes in slationary phase,which are synthesized by complex secondary metabolic pathways,with simple structure,clear metabolic pathways and large yield of primary metabolites as precursors.At present,a large number of studies have shown that the secondary metabolites of endophytic fungi are rich in species,novel in structure and diverse in biological activity.It is a treasure house for providing bioactive substances for human beings.Our research team is devoted to the study of biodiversity,chemical diversity and host relevance of endophytic fungi in medicinal plants.In the previous study,we isolated and identified the Trichoderma atroviride,which were numbered D16 and S361 respectively,from Salvia miltiorrhiza and Cephalotaxus fortunei.D16 could produce tanshinone I and tanshinone II A,while S361 could not.After consulting the literature,we found that homologous endophytic fungi from other genera in different hosts can produce secondary metabolites similar to the host active components,and we suspect that similar phenomena may occur in secondary metabolites of D16 and S361.In this paper,the endophytic fungus S361 isolated from Cephalotaxus fortunei Hook.f.(grown in Jiande,Zhejiang province)and the endophytic fungus D16 isolated from Salvia miltiorrhiza(grown in Shangluo,Shanxi province)were studied.Their secondary metabolites were isolated and identified,while the biological activities of the compounds were studied.After the fermentation of Strain D16 in rice solid medium for 30 day,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 gel column chromatography and semi preparative HPLC method were used for the fermentation separation and purification.At last,we obtained 17 compounds.Then,we determined their structures through a variety of wave spectral data(NMR,MS,IR.CD),compared with the literature.We found that this fermentation included 3 unknown compounds(3 new sesquiterpenoids)together with 14 known compound(5 esters,2acids,1 phenol,1 alcohol,1 ketone,3sesquiterpenoids,1 diterpenoid).After the fermentation of Strain S361 in rice solid medium for 30 day,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 gel column chromatography and semi preparative HPLC method were used for the fermentation separation and purification.At last,we obtained 16 compounds.Then,we determined their structures through a variety of wave spectral data(NMR,MS,IR,CD),compared with the literature.We found that this fermentation included 4 unknown compounds(2 acid amide,1 sesquiterpenoids,1 diterpenoid),together with 12 known compounds(3 lactones,1 phenol,3 alkaloids,3 sesquiterpenoids,1 diterpenoid,1 quinone).After extensive review of literature,it was speculated that 5 new terpenoids have anti-inflammatory activity.The anti-inflammatory activities of the related compounds were tested by LPS stimulation.The results showed that these new terpenoids have no anti-inflammatory activity.