We isolated 100 Vibrio sp from marine products and sea water 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 Viv\brio sp. showed identical, in...
We isolated 100 Vibrio sp from marine products and sea water 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 Viv\brio sp. showed identical, in eneral V. vulnificus ys-I produced hemolysin as the hightest titer The optimal culture conditions for the hemolysin production by the V. vulnificus ys-I are followings ;
1. Hemolysin production was optimal during 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 mitial pH of 8.0, 30˚C, 3% NaCl.
Hemolysin ws purified from culture filtrate of the strain by ammonium sulfate recipitation, on exchange and hydrophobic interaction chromatography. The results were as follows ;
1. Homogeneity of the purified hemolysin was demonstrated by revealing single band on SDS-PAGE. The molecular weight of purified hemolysin was 45KDa
2. The absorbance pattern in ultraviolet was typical of those seen with most protein with 280nm.
3. Punified hemolysin was stable at 50˚C but 70% of the activity was lost by heating for 30min t 60˚C Optimal temperature of purified hemolysin was 35˚C.
4. Purified hemolysin was stable at the pH range of 6~9, but in the less the pH 5.0, above the pH 9.0, the he-molysin acivity was lost completely.
Key words : Viulence factor, Vibrio vulnificus ys-I, hemolysin purification