Study On The Metabolites From Marine Fungus By The Bioactivity Directional Screening

The bioactivity directional screening technology with dual-mode has been designed in this paper, using HPLC correspond with bioactivity profile to identify biological active components, with a view to finding active natural products from marine-derived fungus. Several main conclusions are drawn as follows:(1) Marine fungus ENP701# which was isolated from seawater sample collected from the East China Sea was analyzed as experimental subjects in this study. It was identified to be Penicillium sp. species based on the morphological evaluation. After large-scale cultivation of marine fungus ENP701#, the mycelium and culture fluid were separated by filtration, and extract were gained, respectively.(2) The volatile components of EtOAc extract from culture fluid of marine fungus ENP701# were analyzed by GC-MS at different culture time. The results revealed that volatile components from EtOAc extract were mainly complex and aromatic compounds, such as long-chain alkanes, alkaloids, coumarin and ester compounds. Marine-derived fungus can produce different volatile components. The volatile components from marine- derived fungus were different at the different culture time.(3) The bioactivity directional screening technology with dual-mode of mycelium and culture fluid extract has been established to provide referrence for follow-up seperation and purification of compounds, with a view to screening active natural products from metabolites of marine- derived fungus. During establishment of anti-tumor activity directional screening model, six tumor cell lines including H460, PC-12, A549, 3T3, HT1080 and Hela were selected in order to screen appropriate cell lines. The anti-oxidative activity directional screening model was set up based on DPPH free radical scavenging detection. The bioactivity directional screening with dual-mode of mycelium and culture fluid extract were established mainly on H460 cell inhibition, assisted by DPPH radical scavenging detection profiles eventually.(4) Based on the bioactivity directional screening spectrum with dual-mode of mycelium and culture fluid extract, six target bioactive compounds were directly isolated from the mycelium extract with middle pressure preparation instrument according to the best conditions of HPLC. They were Choline-O-Sulphate(1), 1,2,3,4,5,6-hexanehexanol(2), 1,2,3,4- erythritol(3), complex quinoline alkaloid(4), 5,7,22- triene-3β-hydroxy- ergosterol(5), stearic acid (6), respectively. Compounds 4 was a new compound, it showed significant inhibitory activity against H460 tumor cells with IC₅₀ value of 19.38±0.15μg·mL^-1. In addition, all the compounds revealed scavenging activity on DPPH free radical so that compounds 1-6 had different degrees of anti-oxidative activity.