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We isolated 100 Vobrio sp. from marine products and sea from July to September, 1997. We attemped on purification of hemolysin produced by Vibrio sp. The growth, hemolysin production patterns by the 100 strains of Vibrio sp. showed identical, in general. V. unlnificus ys-1 produced hemolysis as the higtest titer. The optimal culture conditions for the hemolysin production by the V. vunificus ys-1 are followings; 1. Hemolysin production was optimal dering the late exponetial phage. 2. Maximal growth, hemolysin production were in heart infusion broth. 3. Maximal yields of hemolysin was obtained when the heart infusion broth had an intial pH of 8.0, 30℃, 3% NaCL. Hemolysin was purified from culture filtrate of the strain by ammonium sulfate recipitation, ion exchange and hydrophobic interaction chromatography. The results were as follows; 1. Hemogeneity of the purified hemolysin was demonstrated by revealing single band on SDS-PAGE. The molecular weight of purified hemolysin was 45KDa. 2. The absorbance rattern in ultraviolet wsa typical of those seen with most proteinb with 280nm. 3. Purified hemolysin was atable at 50℃ but 70℃ of the acivity was lost by heating for 30 min at 60℃/ Optimal temperature of purified hemolysin was 35℃. 4. Purified hemolysin was stable at the pH range of 6∼9, but in the less the pH5.0. above the pH 9.0, the hemolysin activity was lost completely.
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