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用于快速灵敏检测饮用水中汞的 MerR-荧光蛋白嵌合体生物传感器。

MerR-fluorescent protein chimera biosensor for fast and sensitive detection of Hg in drinking water.

机构信息

Department of Biochemistry, Faculty of Science, Ege University, Bornova-Izmir, 35100, Turkey.

Department of Chemistry and Chemical Processing Technologies, Lapseki Vocational School, Canakkale Onsekiz Mart University, Canakkale, Lapseki, Turkey.

出版信息

Biotechnol Appl Biochem. 2019 Sep;66(5):731-737. doi: 10.1002/bab.1805. Epub 2019 Aug 30.

Abstract

Mercury ion (Hg ) is a universal pollutant and its detection is crucial for public healthcare. In this study, we developed a novel fluorescent biosensor by construction of a protein fusion between the mercury-sensing transcription factor MerR and enhanced yellow fluorescent protein (EYFP). Hg -induced conformational change of MerR was transduced into fluorescence signal. Fluorescence intensity of the biosensor protein decreased with increasing concentrations of Hg and a linear response was obtained in the range of 0.5-40 nM. The limit of detection was 0.5 nM, which was much lower than the maximum allowed level in water. The biosensor specificity was highly dependent on type and concentration of metal ion. The biosensor exhibited high specificity in a mixture of metal ions at 0.5 nM concentration. However, the interference effect was more pronounced at 40 nM concentration of metal ions. The measurement was completed in less than 1 Min with no need for sample preparation or preincubation steps. The biosensor achieved accurate and reliable detection in the spiked drinking water sample, as validated by the inductively coupled plasma optical emission spectrometry.

摘要

汞离子(Hg )是一种普遍存在的污染物,其检测对公共卫生健康至关重要。在本研究中,我们通过构建汞感应转录因子 MerR 和增强型黄色荧光蛋白(EYFP)的蛋白融合,开发了一种新型荧光生物传感器。Hg 诱导的 MerR 构象变化被转化为荧光信号。生物传感器蛋白的荧光强度随 Hg 浓度的增加而降低,在 0.5-40 nM 范围内得到线性响应。检测限为 0.5 nM,远低于水中允许的最高水平。生物传感器的特异性高度依赖于金属离子的类型和浓度。在 0.5 nM 浓度的混合金属离子中,该传感器表现出高度的特异性。然而,在 40 nM 浓度的金属离子下,干扰效应更为明显。测量在不到 1 分钟内完成,无需样品制备或预孵育步骤。该生物传感器在经过电感耦合等离子体光学发射光谱法验证的加标饮用水样品中实现了准确可靠的检测。

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