Isolation Of Endophytes Fungi From Five Plants,and Testing Their Secondary Metabolites For MAO And AChE Inhibitory Activity

Plant endophytic fungi are defined as the fungi which spend the whole or part of their lifecycle colonizing inter-and/or intra-cellularly inside the healthy tissues of the host plants,typically causing no apparent symptoms of disease.Monoamine oxidase(MAO)enzymes are drug targets for antidepressants(which inhibit the MAO-A isoform)and for anti-Parkinsonian(which inhibit the MAO-B isoform)agents.Acetylcholinesterase is an enzyme that catalyzes the breakdown of acetylcholine and of some other choline esters that function as neurotransmitters,the inhibitors of this enzyme are used to treat Alzheimer's disease.In this research,35 strains of endophytic fungi have been isolated from five Chinese medicinal plants.11 strains were isolated from Iris alctea(??),7 strains were isolated from Desmodium elegans(?????),6 strains were isolated from Salvia miltiorrhiza(??),6 strains were isolated from Pedicularis resupinata(???)and 5strains were isolated from Parasenecio ambiguus(?????).The strains were fermented and extracted with ethyl acetate and methanol that afforded 70 extracts.The inhibitory activity of 70 extracts against monoamine oxidase and acetylcholinesterase were also tested.The strain SMH-3 isolated form Desmodium elegans(?????)showed significant inhibitory activity against monoamine oxidase with IC50 value of 16.2?g/mL,whereas the others strain of isolated fungi were showed no or weak inhibitory activity against monoamine oxidase and acetylcholinesterase.The chemical composition of this strain SMH-3 was investigated by modern chromatographic methods,including preparative HPLC,TLC,silica gel column chromatography.8 compounds were isolated and identified according to their H-NMR,C-NMR and MS spectral data as flufuran(1),deactylfructigenines A(2),(N-(4-hydroxymethoxy phenyl)acetamide(3),fructigenines B(4),Ergosterol(5),Ergosterol peroxide(6),Epi-aszonalenin A(7).Dibutyl phathalate(8).Compound 1(flufuran)showed a significant anti-monoamine oxidase activity with an IC50 value of 16.2 ?M when the IC50 value of iproniazid as standard was 5.7?M.By cluster analysis,morphology and color of mycelium colony,and observation of conidia,The strain SMH-3 was identified as Penicillium polonicum.