Study On The Active Components Of Marine Fungi MNP801 By Activity-based Fingerprint Technology

This research designated activity by molecular recognition technology,that technology by HPLC combined with antioxidant activity or anti-tumor activity screening techniques,preliminary determined the polar regions of marine fungi extract's activity part,thus the separation of metabolites of marine fungi in the process of providing certain guidance,according to this,secondary metabolites of marine fungi were separated and purified systematically,and finally obtained bioactive metabolites of marine fungi,as a way of screening active ingredients from metabolites of marine fungi.Several main conclusions are drawn as follows:(1)The experiment strain MNP801 were isolated from the water sample which was collected in latitude 31.3622 degrees,east longitude 122.6896 degrees 40 meters deep sea of the East China Sea by multi-diluted coating method and strains of marine fungus.One of them was identified by 18sDNA genus,it was Alternaria sp.MNP801,cultived with large-scale,separated cell and fermentation liquid,extracted the cell contaits.The other strains identified marine fungus Mucor sp.MNP801 by the initial biochemical and morphological way,with its conduct specific biotransformation of natural products.(2)By HPLC combined with its antioxidant activity or anti-tumor activity screening technology to build the fixed activity spectrum of marine fungi Alternaria sp.MNP801.In the antioxidant model,mainly by measuring the DPPH· radical scavenging activity to evaluate compounds of each section antioxidant activity of the polar.In the anti-tumor sentinel activity model,respectively,measured the inhibitory activity to PC-12,H460,A549 and 3T3 these four tumor cells,after screening found that inhibitory activity to H460 and 3T3 tumor cells is relatively significant,and finally isolated the antitumor activity compounds basis for inhibition of 3T3 and H460 cell line activity.(3)Based on the established antioxidant and anti-tumor sentinel identification of molecular spectra,by determining the optimal HPLC conditions,used of the Suppression Equipment Instrument directly preparation the target trace compounds of cell extraction by the fixed-point separation,and ultimately isolated compounds 1-3,respectively,5 a,8 a-epidioxy-ergosta-6,22-diene-3 ?-ol(1),xanthone(2),Stearic acid(3)from the cell extraction.Compound 1 was the first time separated from the marine fungus Alternaria sp.,showed some inhibitory activity to the H460 and 3T3 tumor cells,its IC5O value of this compound was 51.26± 0.21 ?g·mL-1 and 41.26 ± 0.18?g·mL-1,respectively.(4)Biotransformated the natural product 6,7-dimethoxy coumarin which isolated from willow bloom by marine fungi Mucor sp.MNP801,used of GC-MS to analyze the fermented liquid's volatile components of the transform group and the control group,One new product,2-Acetyl-4-acetamido-5-nitrophenol,was found in transformation group without the appearance in control group,it was not the secondary metabolites that produced by ths through e marine fungus itself,provides a way to obtain new activity substancemarine fungi.