Chemical Constituents Of Endophytic Fungi P.chrysogenum MT-12 And A.alternate MT-3.970 From Huperzia Serrat

Endophyte is a fungus or bacterium that lives in the tissues and organs of healthy plants in a certain stage or all stages,and could excit in the plant’s roots,stems,leaves,flowers,fruits and seeds.Due to the difference of host plants and their living environment,endophytes are diverse in species,large in number and rich in metabolites.Huperzia serrata is a perennial fern belonging to the family Huperzia Bernh,which has effects of contusions,swellings,alleviating pains and detoxificationg.Our group has identified 46 endophytic fungi from H.serrata,including the 12 familys of Glomerella sp.,Colletotrichum sp.,Penicillium sp.,Arthrinium sp.,Sphaerulina sp.,Cladosporium sp.,Mycosphaerella sp.,Cercospora sp.,Mucor sp.,Alternaria sp.,Malassezia sp.and Podospora sp..In this study,the crude extracts of 40 endophytic fungi metabolites were screened in vitro,crude extracts of two strains of endophytes,Penicillium chrysogenum MT-12 and Alternaria alternate MT-3.970,with significant activity were selected.Accordingly,an ongoing study for screening potential activity agents from natural products,the metabolites produced by P.chrysogenum MT-12 and A.alternate MT-3.970,endophytic fungus isolated from Chinese club moss H.serrata,were investigated.Their structures were elucidated on the basis of spectroscopic analysis,including spectroscopic and spectrometric data(1D and 2D NMR,IR,and HRESIMS).24 new compounds,along with 18 known compounds were isolated from the solid substrate fermentation cultures of P.chrysogenum MT-12 named chrysogenolides A-H(P1*-P8*),berkeleyacetal A(P9),berkeleyacetal B(P10),berkeleyacetal C(P11),purpurogenolide C(P12),22-epoxyberkeleydione(P13),berkeleyd-ione(P14),and berkeleyone B(P15),penicichrysogenins A-L(P16*-P26*),asperlone A(±),(P27),4,6-dihydroxy-5-methylphthali-de(P28),6-hydroxy-4-methoxy-5-met-hylphthalide(P29),and 5-carboxymethyl-2-heptyl-7hydroxychromone(P30),penicichr-ysogenin M(P31a*),penicichrysogenin N(P31b*),Isopenicillide(P32),Penicillide(P33),Talaromyone A(P34),hydroxypenicillide(P35),purpactin A(P36),penicillide D(P37),penicichrysoge-nilliden M(P38),penicichrysogenilliden N(P39).Compounds P1-P8,P16-P26,P31a/P31b,P38,and P39 were new compounds.On the biological activities of the compounds P3,P4,P6,P11,P12,P19,P20,P23,P24,P26a,P26b,P31a,and P31b from P.chrysogenum MT-12 show exhibited inhibition of nitric oxide production in lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cells with IC50 values in the range of 4.3-98.4 μM.P31a,P31b,and P33 induced apoptosis of HepG2 cells with IC50 values in the range of 15.0-95.4 μM.In contrast,all the compounds didn’t exhibit inhibitory activities on AChE at the concentration of 100 μM.In addition,the biosynthetic mechanisms of these compounds from P.chrysogenum MT-12 were disscussed.Three new compounds(Al*-A3*),along with 12 known compounds(A4-A15)were isolated from the solid substrate fermentation cultures ofAlternaria alternata MT-3.970,named pencialternata acid A(Al),pencialternata acid B(A2),pencialternata acid C(A3),(+)altennuene-2-acetoxy ester(A4),(+)altennuene-3-acet-oxy ester(A5),altenuisol(A6),2,5dimethyl-7-hydroxychromone(A7),(-)alternarlactam(A8),1-deoxyrubralactone(A9),1,6dihydroxy-8-methoxy-3a-methyl-3,3a-dihydrocyclopen-ta[c]isochromene-2,5-dione(A10),alternariol 9-methyl ether(All),alternariol(A12),xanalteic acid Ⅱ(A13),stemphytriol(A14),alterperylenol(A15).Compound A7 showed inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophage cells with IC50=51.5 ±1.3 μM.All compounds from A.alternata MT-3.970 show any activities whitch them had screen for P.chrysogenum MT-12.In addition,the biosynthetic pathways for the compounds from A.alternata MT-3.970 were disscussed.