The hemolysin of Vibrio cholerae non-O1 CT isolated from sea water was purified and characterized. The purified hemolysin displayed an optimum at $37^{circ}C$ and exhibited more than 70% of residual hemolytic activity after incubation at $45^{circ}C$ for 120 min. However, the activity dropped dramatically at temperature above $55^{circ}C$. The purified protein showed the highest hemolytic activity at pH 7.0, while the activity was completely lost outside of the pH ranges of 5.0 and 10.0. The activity of hemolysin was inactivated by addition of divalent cations, such as $Cu^{2+},;Fe^{2+},;Hg^{2+},;Mn^{2+},;and;Zn^{2+}$, however, the activity was not completely inhibited by additions of $Ca^{2+},;Mg^{2+},;K^+,;Na^+,;and;Li^+$.