Abstract

Vibrio cholerae non-O1 FM-3 was isolated from sea water, and it showed the same bacteriological characteristics as Vibrio cholerae non-O1 ATCC 25872. V. cholerae non-O1 FM-3 presented the highest hemolytic activity at stationary phase of its growth. The hemolytic activity was decreased in accordance with increasing of pretense activity of its cultural supernatant. The characteristics of the hemolysin produced by V. cholerae non-O1 FM-3 were investigated after partial purification with a Sephadex G-100 chromatography. The hemolytic activity of purified protein was stable at $4^{\circ}C$ while it was completely lost by heating at $60^{\circ}C$ for 30 min. The activity of hemolysin was increased by addition of divalent cations such as $Ca^{2+},\;Mg^{2+},\;and\;Mn^{2+}$ while it was inhibited by additions of $Zn^{2+}$. When the hemolysin was incubated with suspensions of erythrocytes at $4^{\circ}C$ and $37^{\circ}C$, respectively, hemolysis was not observed at $4^{\circ}C$ but at $37^{\circ}C$. It means that hemolysis by purified hemolysin was temperature-dependent while its binding step of hemolysin to cell membrane was temperature-independent.

해수로부터 분리한 V. cholerae non-O1 FM-3 이 생산하는 hemolysin의 특성에 관하여 연구한 결과를 요약하면 다음과 같다. 1. V. cholerae non-O1 FM-3이 생산하는 hemolysin은 대수증식기일 때 가장 높은 활성을 나타내었으며, Sephadex G-100 gel column chromatography로 부분정제한 후 hemolysin의 특성을 실험한 결과, pH 7.0, $4^{\circ}C$에서 가장 안정하였으며, $60^{\circ}C$, 30분 가열에 의해 완전히 실활되었다. 2. 적혈구와 hemolysin이 반응할 때, $Na^+,\;K^+$과 같은 1가 양이온을 첨가하면 약간의 활성증가를 보였으며, $Ca^{2+},\;Mg^{2+},\;Mn^{2+}$과 같은 2가 양이온을 첨가하였을 때 급격한 활성증가를 보였고, 반면 $Zn^{2+}$ 첨가시에는 급격히 활성이 저하되었다. 3. 적혈구 현탁액과 hemolysin을 각각 다른 온도대에서 반응시킨 결과, $4^{\circ}C$에서도 온도와 관계없이 적혈구막과 hemolysin의 결합이 가능한 온도 비의존적인 제1단계와, 용혈황성의 최적온도대인 $37^{\circ}C$에서만 용혈작용이 일어나는 온도 의존적인 제2단계로 나누어졌다.

Keywords

• hemolysin;
• hemolytic activity;
• erythrocytes