| Craniella australiensis belongs to the Demospongiae class, Spirophorida order, Tetillidae family, Craniella genus?Studies have shown that the most chemical composition of the sponge with a novel structure and notable activity,have anti-tumor, anti-bacterial, anti-inflammatory, blood pressure and other biological activity. The two parts of content in this study are as follows:1. The research of chemical composition. C. australiensis was exhaustively extracted with 70% hot ethanol. And then the obtained extract was washed successively with petroleum ether, ethyl acetate, and n-butanol in turn to obtain 3 fractions. By a variety of separation methods and recrystallization methods, three compounds were isolated from the ethyl acetate layer extract and the n-butanol layer extract. On the basis of chemical evidence and spectral analysis, especially by NMR techniques, the struetures of three compounds were elucidated:Aldisin and 2-Bromoaldisin. Both were isolated from the genus for the first time.2. The research of biological activity. All of the layers extract had inhibitory activity of ACE in a dose-dependent manner. the ethyl acetate layer extract was best(IC50 2.61 mg/m L).The antibacterial activities of different layers extract from C. australiensis were studied. The results showed that petroleum ether layer extract had the antibacterial activity with the Bacillus subtilis. The total layer,the ethyl acetate layer and the n-butanol layer extract had a certain inhibitory activity with Shewanella.By measuring the clearance rate of DPPH?,the antioxidant activity of all layers extract were studied. The results showed that all of the total layer,the ethyl acetate layer and the n-butanol layer extract had the ability toclear the DPPH?,and the clearance rates were 90% with 2mg/m L.The inhibition on tumor cells of both compounds from C. australiensis evaluated by CCK-8 assay. The results showed the total layer extracts with 200mg/m L exhibited the potent inhibitory activities against AGS,Hep G2 and SKOV3.The inhibition on tumor cells of both compounds from C. australiensis evaluated by CCK-8 assay. The results showed both Aldisin and 2-Bromoaldisin exhibited the potent inhibitory activities against Caco-2, HCT116, 7721, U251 and A495 cells. The Aldisin showed the strongest cytotoxic activity against 7221 cell with IC50 67.56±3.06 ?g/m L.Both the compounds as natural products could be considered as potential anticancer candidate for further study.Perinereis aibuhitensis belongs to Polychaete class, Nereidida order, Nereidae family,Perinereis gunes. Studies have shown that P. aibuhitensis contain rich protease, whose biological activity was significantly. Many parts of content in this study are as follows:The crude enzyme was obtained from the P. aibuhitensis by ammonium sulfate precipitation. By ion-exchange chromatography on DEAE-52, gel filtration chromatography on sephadex G-100 when bioassay simultaneously, the crude glucosidase was obtained from the crude enzyme. The yield of ?-glucosidase, ?-galactosidase, ?-mannosidase were 39.5%, 20.3% and 1.9%.The effect of the temperature, p H, metal ions to the enzyme activity, as well as enzyme kinetics of glucosidase were studied.Studies have shown that ?-glucosidase exhibited optimum activity at p H 7 and 50?. The Km and Vmax of ?-glucosidase were 8.14×10-5 M and 3.19×10-4 M/h. ?-galactosidase exhibited optimum activity at p H 6 and 50 ?. The Km and Vmax of ?-glucosidase were 1.20×10-4 M and 1.63×10-4 M/h. ?-mannosidase exhibited optimum activity at p H 4 and 70?. The Km and Vmax of ?-glucosidase were 6.17×10-5 M and 1.82×10-5 M/h. The enzyme activity was inactivated different degrees by glycosidases, especially Ag+. |
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