碩士 / 國立海洋大學 / 水產養殖學系 / 87 / The present study is aimed to (1) investigate the interactions between extracellular products (ECP) of pathogenic vibrios and serum components of grouper ; (2) partially purify the protease inhibitor in the grouper serum and (3) discuss its possible role on vibriosis.
Grouper serum contains one protease inhibitor which can inhibit ECP protease activity of Vibrio alginolyticus. It could inhibit activity of serine protease but not that of cysteine protease. This protease inhibitor was thermal resistant . After incubation at 65℃ for 20 minutes, it still contained 75﹪inhibitory capacity. It was not sensitive to methylamine.
The protease inhibitor was partially purified by ammonium sulfate precipitation、isoelectric focusing and anion exchange chromatography. After Mono-Q anion exchange chromatography separation , fractions which had inhibitory capacity towards V. alginolyticus ECP protease contained two dominant protein bands in a native-PAGE gel , one band’s molecular weight was about 67 KDa , and the other’s was lower than 67 KDa. By the way of zymogram in native-PAGE , the band with molecular weight about 67 KDa was demonstrated to have inhibitory capacity towards V. alginolyticus protease. The partially purified protease inhibitor contained two distinct bands with molecular weight between 30∼43 KDa and 43∼67 KDa in a SDS-PAGE gel , respectively , and the later was assumed to have inhibitory capacity towards V. alginolyticus protease. Two major precipitation arcs were visualized in partially purified grouper serum in a crossed immunoelectrophoresis (CIE) gel using rabbit antiserum to the normal whole grouper serum.
No matter if grouper serum could inhibit ECP’s protease activity, there was one specific grouper serum component markedly affected and had fast-migration in a CIE gel following in vitro ECP treatment. After incubation with V. alginolyticus ECP , the grouper’s partially purified protease inhibitor could inhibit ECP protease activity , but this inhibitor did not perform fast-migration in a CIE gel. It was therefore believed that the specific grouper serum component which had fast-migration in a CIE gel after incubation with V. alginolyticus ECP in vitro was not the protease inhibitor in grouper serum.
| Identifer | oai:union.ndltd.org:TW/087NTOU0086030 |
| Date | January 1999 |
| Creators | WeiMing Su, 蘇偉銘 |
| Contributors | Lee Kuo-Kau, 李國誥 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 0 |
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