Basic Research on the Development of Detoxification Techniques of Endocrine Disrupting Chemicals, Short Ethoxy Chain-Alkylphenols

内分泌干​​扰物短乙氧基链烷基酚解毒技术发展的基础研究

• 批准号: 19780240
• 负责人: TASAKI Yuji
• 金额: $ 1.8万
• 依托单位: Nagaoka National College of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19780240/
• 关键词: 環境; 水資源; 遺伝子; 応用微生物; 酵素; バイオテクノロジー; 農薬補助剤; アルキルフェノールポリエトキシレート; 微生物分解; 内分泌かく乱物質; トランスポゾンタギング; Pseudomonas属細菌

Alkylphenol polyethoxylates (APEOs) belonging to a group of nonionic surfactants are biodegraded by bacteria and then short ethoxy chain-alkylphenols which are potential endocrine disrupters are produced. In this study, to elucidate the biodegradation mechanism of APEOs, we isolated genes that were involved in biodegradation of octylphenol polyethoxylate (OPEO), a kind of APEOs, from OPEO degrader Pseudomonas putida S-5 by transposon tagging and analyzed their DNA sequences. Approximately 11,000 transposon mutants were constructed by insertion of transposon Tn5 into P.putida S-5. Seventeen mutants that are not able to metabolize OPEO, namely OPEO-degradation gene disruptants, were isolated. DNA sequences of the genes disrupted by insertion of transposon were determined. A similarity search showed that 17 proteins encoded by the genes have high amino acid sequence similarity to acyl-CoA synthetase, chaperone protein, NADH quinone oxidoreductase, etc. Gene expression of 9 genes in P.putida S-5 cells grown on different carbon source (OPEO or glucose) was analyzed by RT-PCR. T424 gene was expressed only in cells grown on OPEO, while the other genes were expressed in cells grown on glucose as well as on OPEO.

烷基酚聚氧乙烯醚(APEO)是一类非离子表面活性剂，被细菌生物降解后产生短链烷基酚类物质，具有潜在的内分泌干扰作用。本研究利用转座子标签技术，从恶臭假单胞菌S-5菌株中分离了与辛基酚聚氧乙烯醚降解相关的基因，并对其进行了序列分析，以阐明APEO的生物降解机理。通过将转座子Tn5插入恶臭假单胞菌S-5，构建了约11,000个转座子突变体。筛选出17株不能代谢OPEO的突变体，即OPEO降解基因干扰物。测定了转座子插入打乱基因的DNA序列。相似性检索表明，这些基因编码的17个蛋白质与酰辅酶A合成酶、伴侣蛋白、NADH苯醌氧化还原酶等氨基酸序列具有较高的相似性。T424基因仅在OPEO上表达，其余基因在葡萄糖和OPEO上均有表达。

项目成果

Pseudomonas putida S5株のアルキルフェノールポリエトキシレート分解遺伝子の単離と解析

恶臭假单胞菌S5菌株烷基酚聚氧乙烯醚降解基因的分离与分析

• 发表时间: 2010
• 作者: 高橋一也;山谷真也;大山雄輝;長井隆;田崎裕二
• 通讯作者: 田崎裕二

Pseudomonas putida S-5のオクチルフェノールポリエトキシレート分解遺伝子の塩基配列と発現の解析

恶臭假单胞菌S-5辛基酚聚氧乙烯醚降解基因核苷酸序列及表达分析

• 发表时间: 2008
• 作者: 高橋一也;山谷真也;大山雄輝;長井隆;田崎裕二;田崎 裕二
• 通讯作者: 田崎 裕二

非イオン系界面活性剤分解菌モニタリングのためのアルコールデヒドロゲナーゼ遺伝子を指標とした分子デバイスの設計

以乙醇脱氢酶基因为指标监测非离子表面活性剂降解菌的分子装置设计

• 发表时间: 2008
• 作者: 小栗まゆみ;松尾賀津代;細田晃文;田崎裕二;吉川博道;田村廣人
• 通讯作者: 田村廣人