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首页> 外文期刊>Science of the total environment >Evaluation and validation of a novel Arxula adeninivorans estrogen screen (nAES) assay and its application in analysis of wastewater, seawater, brackish water and urine
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Evaluation and validation of a novel Arxula adeninivorans estrogen screen (nAES) assay and its application in analysis of wastewater, seawater, brackish water and urine

机译：新型Arxula adeninivorans雌激素筛查（nAES）分析的评估和验证及其在废水，海水，微咸水和尿液分析中的应用

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摘要

A novel Arxula adeninivorans yeast estrogen screen (nAES) assay has been developed for detection of estrogenic activity in various liquid samples such as wastewater, seawater, brackish water and swine urine. Two bio-components were engineered to co-express the human estrogen receptor α (hERα) and an inducible reporter gene; either the non-conventional phytase gene (phyK, derived from Klebsiella sp. ASRI) or the non-conventional tannase gene (ATAN1, derived from Arxula). Both reporters were put under the control of an Arxula derived glucoamylase (CAA) promoter, which was modified by the insertion of two estrogen-responsive elements (EREs). The Arxula transformation/expression platform Xplor? 2, which lacks resistance markers and E coli elements, was used to select stable mitotic transformants. They were then analyzed for robustness and suitability as the bio-component for the nAES assay. Two types of the nAES assay based on the reporter proteins phytase and tannase (nAES-P, nAES-T) were used in this work. The nAES-P type is more suitable for the analysis of seawater, brackish water and urine whereas the nAES-T type exhibited higher robustness to NaCl. Both assay types have similar characteristics for the determination of estrogen in sewage and urine samples e.g. 6-25 h assay period with detection and determination limits and EC_(50) values for 17β-estradiol of 2.8 ng L~(-1) 5.9 ng L~(-1) 33.2 ng L~(-1) (nAES-P) and 3.1 ng L~(-1) 6.7 ng L~(-1) and 39.4 ng L~(-1) (nAES-T). Substrate specificity and analytical measurement range (AMR) for both assay types are also similar. These characteristics show that the nAES assay based on non-conventional salt tolerant yeast is applicable for a high throughput estrogen analysis in the environmental and regulatory control sectors.
机译：已经开发出一种新颖的Arxula adeninivorans酵母雌激素筛选（nAES）方法，用于检测各种液体样品（如废水，海水，微咸水和猪尿）中的雌激素活性。工程改造了两个生物成分来共同表达人类雌激素受体α（hERα）和诱导型报告基因。非常规植酸酶基因（phyK，源自克雷伯菌属ASRI）或非常规鞣酸酶基因（ATAN1，源自Arxula）。两个记者都置于Arxula衍生的葡糖淀粉酶（CAA）启动子的控制下，该启动子通过插入两个雌激素响应元件（ERE）进行了修饰。 Arxula转换/表达平台Xplor？缺少抗性标记和大肠杆菌元素的图2用来选择稳定的有丝分裂转化子。然后分析它们的稳健性和适用性，作为nAES分析的生物成分。在这项工作中使用了两种基于报告蛋白植酸酶和鞣酸酶的nAES分析方法（nAES-P，nAES-T）。 nAES-P型更适合于分析海水，微咸水和尿液，而nAES-T型对NaCl表现出更高的耐用性。两种测定类型对于测定污水和尿液样品中的雌激素具有相似的特性，例如：具有检测和测定极限的6-25小时检测周期以及2.8 ng L〜（-1）5.9 ng L〜（-1）33.2 ng L〜（-1）的17β-雌二醇的EC_（50）值（nAES-P ）和3.1 ng L〜（-1）6.7 ng L〜（-1）和39.4 ng L〜（-1）（nAES-T）。两种测定类型的底物特异性和分析测量范围（AMR）也相似。这些特征表明，基于非常规耐盐酵母的nAES分析适用于环境和监管部门的高通量雌激素分析。

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  • 来源
    《Science of the total environment》 |2010年第23期|p.6017-6026|共10页
  • 作者

    Christian Kaiser; Steffen Uhlig; Torsten Gerlach; Martina Koerner; Kirsten Simon; Kirstin Kunath; Kristina Florschuetz; Keith Baronian; Gotthard Kunze;

  • 作者单位

    Leibniz Institute of Plant Genetics and Crop Plant Research (1PK), Corrensstr. 3, D-06466 Catersleben, Germany;

    quo data GmbH, Kaitzer-Str. 735, D-01187 Dresden, Germany;

    Leibniz Institute of Plant Genetics and Crop Plant Research (1PK), Corrensstr. 3, D-06466 Catersleben, Germany;

    Leibniz Institute of Plant Genetics and Crop Plant Research (1PK), Corrensstr. 3, D-06466 Catersleben, Germany;

    quo data GmbH, Kaitzer-Str. 735, D-01187 Dresden, Germany;

    quo data GmbH, Kaitzer-Str. 735, D-01187 Dresden, Germany;

    Leibniz Institute of Plant Genetics and Crop Plant Research (1PK), Corrensstr. 3, D-06466 Catersleben, Germany;

    School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand;

    Leibniz Institute of Plant Genetics and Crop Plant Research (1PK), Corrensstr. 3, D-06466 Catersleben, Germany;

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  • 正文语种 eng
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  • 关键词

    arxula adeninivorans; endocrine disruption; estrogen receptor; reporter assay; urine; yeast assay;

    机译：Arxula adeninivorans;内分泌干​​扰雌激素受体报告基因检测尿;酵母测定;

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