Purification,Characteristics And Bioactivity Of Antiviral Protein From The Pleurotus Eryngii And Agrocybe Sp.

A protein, named yy4a with anti-TMV (Tobacco mosaic virus) activity, was isolated and purified to homogeneity from the fruiting bodies of edible fungus maitake (Yunyougu sp.) by (NH4)2SO4 precipitation followed by DEAE -Sepharose FF and Sephacryl S-200 High Resolution column chromatography. The molecular weight of the protein is about 17 kDa by SDS-PAGE. The lowest inhibitory concentration of yy48 was 3.13 ug/mL when Nicotiana gluiinosa was infected by TMV with the density of 20 ug/mL. It displayed high haemagglutinating activity, which was inhibited by lactose and inulin. When the initial concentration was 1.5 mg/mL, it's liter of agglutinating rabbit erythrocytes was 2-9. yy4a also exhibited high agglutinating activity towards HL-60 and a potent inhibitory activity on HL-60 with an IC50 value of 0.2 mg/mL.An antiviral protein, named XB, was isolated and purified from the fresh edible mushroom, Pleurotus eryngii, through two seriate chromatographic steps: DEAE-Sepharose FF and then Superdex75 Prep Grade .The protein had a molecular weight of 22.3 kDa and showed homogeneity on the native polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing electrophoresis (IEF). XB was an acidic monomer containing 0.4% sugar with an isoelectric point of 4.1. It appeared in pileus more than in stipe. Antiserum, with 2-13 titer and high speciality detected by enzyme-linked immunosorbent assay (ELISA) and western immunoblot, was prepared by immunizing rabbit with XB, which was further purified by SDS-PAGE. It also exerted a haemagglutinating activity on rabbit erythrocytes. The titer was 2-3 when the start concentration was 0.075 mg/mLThe XB protein had a lowest inhibition rate against TMV when its concentration was 4.69 ug/mL. XB did not affect multiplication of TMV in infected leaf discs and did not show distinctly reducing infectivity when applied either before or after TMV inoculation. By observation of TMV treated with XB throughout the electron microscope. XB adhered to TMV viron uniformly. The PAGE showed that XB binded TMV coat protein. It was proposed that XB inhibit TMV CP by binding with TMV to reduce TMV activity or to interfere withrecognition site for viral coat protein.We studied the inhibitory action of XB on HL-60 primarily. The result showed that the inhibitory activity exerted in a dose-depend manner and XB also exhibited an increasingly activity with the time passing. It was proposed that the antitumor mechanism of XB induce apoptosis of HL-60. Its IC50 was 15.261 ug/mL by MTT. At the concentration of 15.261ug/mL, the inhibitory rate was 43.7% at the fifth day by cell count.