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Studies on the Toxicification Mechanism of Tetrodotoxin-containing Organisms, Especially on the Dynamic State of TTX-related Substances

河豚毒素生物中毒机制特别是河豚毒素相关物质动态研究

基本信息

批准号：
02304025
负责人：
MIYAZAWA Keisuke
金额：
$ 8.06万
依托单位：
Hiroshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

In order to clarify the mechanisms of toxicification of tetrodotoxin (TTX) -containing organisms, distribution of TTX and its related substances in benthic animals and their inhabiting environments (substratums), the optimal culturing condition of TTX-producing bacteria for its production, classification of TTXproducing bacteria by the determination of base sequence of r-RNA were examined. The isolation and characterization of tetrodonic acid (TDA) -like substance, the possible precursor of TTX in living organisms, were also performed. To conduct these experiments, microanalysis methods of TTX-related substances were also developed.1) A presumed precursor of TTX, TDA-like substance, was isolated from a ribbon worm Cephalothrix linearism It is very unstable and easily converted to TTX, having a specific toxicity of 700 MU/mg, and molecular weight of 320, larger than TTX 1 by mass unit. Another assumed precursor was isolated from a toxic xanthid crab Atergatis floridus, pressumed to be a dimer of TTX. 2) To isolate and analyze these unstable precursors, a LC/FAB MS and GC/MS were developed and used. 3) From the sediments of the Pacific Ocean sounding 4000 m and fresh water lakes, some TTX producing bacteria and Ascomycetes fungi were isolated. Some of them were classified by the sequencing the bases of r-RNA. 4) Some optimal conditions of TTX-producing ability of Vibrio alginolyticus, isolated from a toxic starfish Astropecten poplyacanthus, were estimated to be 25 C for temperature and 3 % for salt concentration of culturing medium. 5) TTX and its related substances were newly detected from some benthic invertebrates inhabiting the East China Sea and the coast of South Kyushu area.

为了阐明河豚毒素(TTX)生物的中毒机制、TTX及其相关物质在底栖动物及其栖息环境(基质)中的分布、产TTX细菌的最佳培养条件、通过测定r-RNA碱基序列对产TTX细菌进行分类等进行了研究。还对河豚酸 (TDA) 样物质（活生物体中 TTX 的可能前体）进行了分离和表征。为了进行这些实验，还开发了 TTX 相关物质的微量分析方法。 1) 从丝带蠕虫 Cephalothrix Linearism 中分离出 TTX 的假定前体，即 TDA 样物质。它非常不稳定，很容易转化为 TTX，比毒性为 700 MU/mg，分子量为 320，按质量单位计比 TTX 1 大。另一种假设的前体是从有毒的黄蟹 Atergatis floridus 中分离出来的，推测是 TTX 的二聚体。 2) 为了分离和分析这些不稳定的前体，开发并使用了 LC/FAB MS 和 GC/MS。 3)从太平洋4000 m深的沉积物和淡水湖泊中分离到一些产TTX细菌和子囊菌类真菌。其中一些是通过r-RNA碱基测序来分类的。 4)从有毒海星Astropecten poplyacanthus中分离出来的溶藻弧菌产生TTX能力的一些最佳条件估计为温度25℃和培养基盐浓度3％。 5)东海及南九州沿岸部分底栖无脊椎动物中新检出TTX及其相关物质。