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Development of bioremediation and of measurement methods of remaining endocrine disrupters in environment.

环境中残留内分泌干扰物的生物修复和测量方法的开发。

基本信息

批准号：
15350091
负责人：
KURASAKI Masaaki
金额：
$ 9.73万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15350091/
关键词：
endocrine disrupters PC12 cell apoptosis bisphenol A nonylphenol tributyltin bax genetic engineering Caspase

项目摘要

Recently, environmental pollutant including the endocrine disrupters has been considered as dysfunctional factor of mutagenesis, and to affect on development and differentiation of fetal. In this study, to confirm the system of bioremediation and measurement methods of endocrine disrupters in environmental by using of genetic engineering, we applied the methods of genetic engineering and analytical chemistry.In this study, on the basis of PC12 cell cultured system which has been known to be induced apoptosis by serum deprivation, we found that tributyl tin inhibited apoptosis and nonylphenol enhanced apoptosis using the cells. These phenomena were considered to be changed the contents of bax and cytochrome C. In addition, we have successful developed convenience measurement system of endocrine disrupters such as dioxins in environment. The detection range of dioxins using the developed assay system showed 1 fmol to 100 pmol.To construct the system of bioremediation of endocrine disrupters, the genes and cDNAs of enzymes that were expected to degrade the endocrine disrupters, has been cloned. The enzymes expressed in Escherichia coli, showed degraded activity of the endocrine. These results obtained in this study are expected to contribute the establishment of bioremediation of remaining endocrine disrupters in environment.

近年来，包括内分泌干扰物在内的环境污染物被认为是诱变的功能失调因素，影响胎儿的发育和分化。本研究运用基因工程和分析化学相结合的方法，确定了利用基因工程技术对环境中内分泌干扰物进行生物修复的体系和测量方法。本研究在已知经血清剥夺诱导凋亡的PC12细胞培养系统的基础上，发现三丁基锡抑制细胞凋亡，壬基酚促进细胞凋亡。这些现象被认为是改变了bax和细胞色素c的含量。此外，我们成功地开发了环境中二恶英等内分泌干扰物的便捷测量系统。该系统对二恶英的检测范围为1fmol ~ 100pmol。为了构建内分泌干扰物的生物修复体系，克隆了有望降解内分泌干扰物的基因和酶的cdna。在大肠杆菌中表达的酶表现出对内分泌的降解活性。本研究结果有望为环境中残留内分泌干扰物生物修复的建立做出贡献。