Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Molecular Science, Shanxi University, Taiyuan 030006, China.
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Molecular Science, Shanxi University, Taiyuan 030006, China.
Spectrochim Acta A Mol Biomol Spectrosc. 2019 Jun 15;217:101-106. doi: 10.1016/j.saa.2019.03.082. Epub 2019 Mar 24.
The development of a simple, rapid and sensitive sensor to detect copper ion is very important for environmental detection. Here, we constructed a fluorescence biosensor based on fluorescence resonance energy transfer (FRET) between copper resistance operon coded protein C (CopC), a copper binding protein, and dansyl chloride (DNS-Cl) to selectively detect copper ion. At alkaline conditions, DNS-Cl was contently attached to CopC forming biosensor of DNS-Cl/CopC, in which fluorescence emission of CopC at 320nm was absorbed by DNS-Cl. After binding with copper ion, the fluorescence of DNS-Cl was quenched significantly to 47%. Within the range of 0.04-11μM (R=0.989), the good linearity was obtained and the detection limit reached to 7nM. More importantly, the biosensor of DNS-Cl/CopC has been successfully used to detecting of copper ion level in purified water, tap water and waste water drained from steel mill. And the results were consistent well with those obtained from the inductively coupled plasma mass spectrometry. Therefore, the established biosensor DNS-Cl/CopC is a creditable method to detect copper ion with high sensitivity and selectivity, which can be utilized as a powerful tool to monitor copper pollution in the environments.
开发一种简单、快速、灵敏的传感器来检测铜离子对于环境检测非常重要。在这里，我们构建了一种基于铜抗性操纵子编码蛋白 C（CopC）与丹磺酰氯（DNS-Cl）之间荧光共振能量转移（FRET）的荧光生物传感器，用于选择性检测铜离子。在碱性条件下，DNS-Cl 与 CopC 充分结合形成 DNS-Cl/CopC 生物传感器，其中 CopC 在 320nm 处的荧光发射被 DNS-Cl 吸收。与铜离子结合后，DNS-Cl 的荧光显著猝灭至 47%。在 0.04-11μM 的范围内（R=0.989），获得了良好的线性关系，检测限达到 7nM。更重要的是，DNS-Cl/CopC 生物传感器已成功用于检测纯化水、自来水和钢厂废水的铜离子水平，与电感耦合等离子体质谱法得到的结果一致。因此，所建立的 DNS-Cl/CopC 生物传感器是一种具有高灵敏度和选择性的检测铜离子的可靠方法，可作为监测环境中铜污染的有力工具。
