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基于有机磷水解酶偶联 UiO-66-NH MOF 的甲基对硫磷高灵敏光学检测。

Organophosphate hydrolase conjugated UiO-66-NH MOF based highly sensitive optical detection of methyl parathion.

机构信息

Nanoscience and Nanotechnology Lab (Division: H-1), Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30C, Chandigarh, 160030, India; Academy of Scientific and Innovative Research (AcSIR-CSIO), Sector 30C, Chandigarh, 160030, India.

Nanoscience and Nanotechnology Lab (Division: H-1), Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30C, Chandigarh, 160030, India.

出版信息

Environ Res. 2019 Jul;174:46-53. doi: 10.1016/j.envres.2019.04.018. Epub 2019 Apr 19.

Abstract

The hexahistidine-tagged organophosphorus hydrolase (OPH) has been immobilized on a Zr-MOF, namely UiO-66-NH. The resulting enzyme-MOF composite was used as a carrier to facilitate the hydrolysis of an organophosphate pesticide, i.e., methyl parathion in to p-nitrophenol (PNP). The formation of PNP took place in direct proportion to the added pesticide concentration. Coumarin1 (7-diethylamino-4-methylcoumarin) was then introduced in the reaction mixture as a reporter fluorescent molecule. As PNP acted to quench the fluorescence of coumarin1, it became possible to detect methyl parathion over a wide concentration range of 10-10 ng/mL with an achievable limit of quantification as 10 ng/mL. The immobilization of OPH on the surface of UiO-66-NH was found to endow an improvement in the enzymatic activity by about 37%. The OPH/UiO-66-NH conjugate was reusable for at least up to eight times and also found stable toward long-term storage (minimum 60 days). The potential practical utility of the above proposed sensing method has been demonstrated by employing it for an accurate analysis of pesticide-spiked food samples.

摘要

已将六组氨酸标记的有机磷水解酶(OPH)固定在 Zr-MOF 上,即 UiO-66-NH。所得酶-MOF 复合物用作载体,以促进有机磷农药(例如甲基对硫磷)水解为对硝基苯酚(PNP)。PNP 的形成与添加的农药浓度成正比。然后,在反应混合物中引入香豆素 1(7-二乙氨基-4-甲基香豆素)作为报告荧光分子。由于 PNP 会猝灭香豆素 1 的荧光,因此可以在 10-10ng/mL 的宽浓度范围内检测到甲基对硫磷,实现的定量限为 10ng/mL。发现 OPH 在 UiO-66-NH 表面的固定化可使酶活性提高约 37%。OPH/UiO-66-NH 缀合物至少可重复使用八次,并且在长期储存(至少 60 天)中也稳定。通过将上述提出的传感方法用于准确分析农药污染的食物样品,已经证明了该方法的实际应用潜力。

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