| Russula vinosa fermentation recipe was optimized by Surface Response Methodology to improve production of lectin from mycelium of Russula vinosa. The optimum culture condition for Russula vinosa was composed of 3.04% starch,0.3% yeast extract,0.2% KH2PO4,0.1% MgSO4, liquid volume 73ml,250 r/min, pH 5, fermentation time 5d. As a result, the total agglutinating activity of intracellular lectin was increased to 72127.3, which was improved 1.88-fold compared to the original conditions.Russula vinosa Lectin (RVL) was isolated and purified from Russula vinosa mycelium by extraction with PB followed by (NH4)2SO4 precipitation, ion-exchange chromatography on DEAE-Sepharose FF and gel filtration on Sephadex G-100. The agglutinic activity of the purified lectin (RVL) increased 73.3 fold and the activity recovery is 54.4%. RVL showed a single protein band on SDS-PAGE and molecular weight of about 55kD was measured when Sephadex G-100 was used.The physico-chemical property and some biologic activity of RVL were studied.lt showed that RVL was a glycoprotein contained of 3.87% neutal sacchaide. Agglutinating activity was detected by rabbit red blood cells, it stabilized at 20-60℃and pH 5-9, and its Agglutinating activity depended on Mn2+,Zn2+ and Ca2+ in some degree. The study on specificity to saccharide showed that D-mannose is the specific inhibition saccharide, sucrose, D-galactose, N-acetyl-D-glucosamine also showed some inhibition to RVL. Amino acid analysis showed that it contained 15 kinds of amino acids,in which the contents of histidine was the highest, but none of arginine and proline.The study on biological activity of RVL showed that it had no effects on tested bacteria,had different effects on some species of tested fungi such as neurospora crassa, Trichoderma viride, Piricularia oryzae and Aspergillus niger, while no inhibitions to spore germination of Aspergillus niger, Trichoderma viride, Aspergillus flavus and Piricularia oryzae. |
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