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Studies on proteinaceous toxins newly found in bivalves belonging to the genus Corbicula

蚬属双壳类新发现的蛋白质毒素研究

基本信息

批准号：
07660267
负责人：
SHIOMI Kazuo
金额：
$ 1.34万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660267/
关键词：
Corbicula japonica Corbicula leana Corbicula sandai Toxicity Proteinaceous toxins タンパク毒

项目摘要

1.All specimens of the brackishwater clam Corbicula japonica obtained from various places of Japan were toxic when their aqueous extracts were intravenously injected into mice. As to toxicity, there were no sexual and seasonal variations but a local variation was obviously recognized. Two other Corbicula species (C.leana and C.sandai) were also toxic, though at much lower levels than C.japonica. Irrespective of the species, viscera were non-toxic and several muscular tissues were toxic ; the highest toxicity was observed in foot muscle for C.japonica, siphon for C.leana and mantle muscle for C.sandai. No toxicity was detected in 12 species of bivalves other than Corbicula species.2.The C.japonica toxin was purified by hydrophobic chromatography, gel filtration and ion-exchange FPLC and shown to be a weakly basic protein with a molecular weight of 25,000 and a pI of 7.7. Its LD_<50> (intravenous injection into mice) was estimated to be 11 mug/kg. The toxin showed hemolytic activity but lacked anti-microbial activity. Although the toxins of C.leana and C.sandai could not be obtained in a pure form, they were comparable to that of C.japonica with respect to stability and molecular weight.3.An attempt was made to improve the time-consuming purification method for the C.japonica toxin described above. As a result, the toxin was purified by only two steps comprising the binding to sheep erythrocyte membranes and ion-exchange FPLC.4.The Corbicula toxins evoke no serious problem to the human health, since they are very unstable to heating and non-toxic upon oral administration. They are of scientific importance in that they are the first examples of proteinaceous toxins from bivalves.

1.对日本各地产的苦咸水蛤（Corbicula japonica）的水提物进行小鼠静脉注射后均有毒性。毒力无性别和季节差异，但有明显的地方性差异。另外两种Corbicula物种（C.leana和C.sandai）也有毒性，尽管水平远低于c.a japonica。无论何种种类，内脏都是无毒的，而一些肌肉组织是有毒的；其中，日本金针菇对足部肌肉、虹吸部位、三台金针菇对套部肌肉的毒性最高。除Corbicula外，12种双壳类动物均未检测出毒性。经疏水层析、凝胶过滤和离子交换FPLC纯化得到的毒素为弱碱性蛋白，分子量为25000，pI为7.7。其LD_<50>（小鼠静脉注射）估计为11马克杯/公斤。毒素具有溶血活性，但缺乏抗微生物活性。3.虽然无法获得赤霉素和三代赤霉素的纯毒素，但其稳定性和分子量与日本赤霉素相当。本文试图改进上述耗时的粳稻毒素纯化方法。结果表明，该毒素仅通过与绵羊红细胞膜结合和离子交换高效液相色谱两步纯化。Corbicula毒素对人体健康没有严重问题，因为它们对加热非常不稳定，口服无毒。它们具有重要的科学意义，因为它们是来自双壳类动物的蛋白质毒素的第一个例子。