首页> 外文期刊>Environmental toxicology and chemistry >DEVELOPMENT OF A HIGHLY SENSITIVE ASSAY FOR ENZYME-MEDIATED REDUCTIVE DEGRADATION OF POLYCHLORINATED DIBENZO-P-DIOXIN
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DEVELOPMENT OF A HIGHLY SENSITIVE ASSAY FOR ENZYME-MEDIATED REDUCTIVE DEGRADATION OF POLYCHLORINATED DIBENZO-P-DIOXIN

机译:酶介导的二苯并-P-二恶英还原降解的高灵敏分析方法的建立

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摘要

The degradation of 2-chloro-4,5-O-(4'-methyl-7', 8'-diphenyl)ether (CMDPE), an analog of 2,7-dichlorodibenzo-p-dioxin (2,7-DCDD), mediated by Geobacillus sp. UZO 3 cell-free extract was monitored. Ethyl acetate extracts of a complete reaction mixture incubated at 65°C for 18 h were analyzed either by thin layer chromatography (TLC) fractionation coupled with spectrometric detection or by gas chromatography-mass spectrometry (GC-MS). The reaction product 4-methylumbelliferone (4MU) was successfully isolated by TLC and visualized by a transilluminator at 450 nm. The 4MU, 4-chlorophenol, and reaction intermediate 6-chlorophenoxy-4-methylumbelliferone were all successfully detected by GC-MS. The presence of these compounds suggest that Geobacillus sp. UZO 3 cell-free extract also catalyzes the reductive cleavage of the diaryl ether bonds of CMDPE in a similar mechanism previously reported in 2,7-DCDD. In the present study, the authors describe a simple and highly sensitive fluorescent assay for a new dioxin degrading enzyme(s).
机译:2-氯-4,5-O-(4'-甲基-7',8'-二苯基)醚(CMDPE)的降解,2,7-二氯二苯并-对二恶英(2,7-DCDD的类似物),由Geobacillus sp。介导。监测UZO 3无细胞提取物。通过薄层色谱(TLC)分离和光谱检测或气相色谱-质谱(GC-MS)分析在65°C下孵育18小时的完整反应混合物的乙酸乙酯萃取液。通过TLC成功分离出反应产物4-甲基伞形酮(4MU),并在450nm下通过透照仪观察。通过GC-MS成功检测到4MU,4-氯苯酚和反应中间体6-氯苯氧基-4-甲基伞形酮。这些化合物的存在暗示了Geobacillus sp.。 UZO 3无细胞提取物还催化CMDPE的二芳基醚键的还原裂解,其机理与先前在2,7-DCDD中报道的类似。在本研究中,作者描述了一种用于新的二恶英降解酶的简单且高度灵敏的荧光测定法。

著录项

  • 来源
    《Environmental toxicology and chemistry》 |2012年第5期|p.1072-1075|共4页
  • 作者单位

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan;

    Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan;

    Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan;

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan;

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan;

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan Takasago Thermal Engineering Co. Ltd., Atsugi, Kanagawa, Japan;

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan;

    Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan;

    Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan;

    Takasago Thermal Engineering Co. Ltd., Atsugi, Kanagawa, Japan;

    College of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa, Japan;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    dioxins; microbial degradation; fluorescent assay; reductive cleavage; diaryl ether bonds;

    机译:二恶英微生物降解;荧光测定还原性裂解二芳基醚键;

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