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原位酶促生成的光开关氧化酶模拟物及其在比色检测葡萄糖氧化酶中的应用。

In Situ Enzymatically Generated Photoswitchable Oxidase Mimetics and Their Application for Colorimetric Detection of Glucose Oxidase.

作者信息

Cao Gen-Xia, Wu Xiu-Ming, Dong Yu-Ming, Li Zai-Jun, Wang Guang-Li

机构信息

The Key Laboratory of Food Colloids and Biotechnology, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, China.

出版信息

Molecules. 2016 Jul 9;21(7):902. doi: 10.3390/molecules21070902.

Abstract

In this study, a simple and amplified colorimetric assay is developed for the detection of the enzymatic activity of glucose oxidase (GOx) based on in situ formation of a photoswitchable oxidase mimetic of PO₄(3-)-capped CdS quantum dots (QDs). GOx catalyzes the oxidation of 1-thio-β-d-glucose to give 1-thio-β-d-gluconic acid which spontaneously hydrolyzes to β-d-gluconic acid and H₂S; the generated H₂S instantly reacts with Cd(2+) in the presence of Na₃PO₄ to give PO₄(3-)-stabilized CdS QDs in situ. Under visible-light (λ ≥ 400 nm) stimulation, the PO₄(3-)-capped CdS QDs are a new style of oxidase mimic derived by producing some active species, such as h⁺, (•)OH, O₂(•-) and a little H₂O₂, which can oxidize the typical substrate (3,3,5,5-tetramethylbenzydine (TMB)) with a color change. Based on the GOx-triggered growth of the oxidase mimetics of PO₄(3-)-capped CdS QDs in situ, we developed a simple and amplified colorimetric assay to probe the enzymatic activity of GOx. The proposed method allowed the detection of the enzymatic activity of GOx over the range from 25 μg/L to 50 mg/L with a low detection limit of 6.6 μg/L. We believe the PO₄(3-)-capped CdS QDs generated in situ with photo-stimulated enzyme-mimicking activity may find wide potential applications in biosensors.

摘要

在本研究中,基于原位形成的磷酸根封端的硫化镉量子点(QDs)的光开关氧化酶模拟物,开发了一种简单且放大的比色测定法,用于检测葡萄糖氧化酶(GOx)的酶活性。GOx催化1-硫代-β-D-葡萄糖氧化生成1-硫代-β-D-葡萄糖酸,后者自发水解为β-D-葡萄糖酸和H₂S;生成的H₂S在Na₃PO₄存在下立即与Cd(2+)反应,原位生成磷酸根稳定的硫化镉量子点。在可见光(λ≥400nm)刺激下,磷酸根封端的硫化镉量子点是一种新型的氧化酶模拟物,通过产生一些活性物种,如h⁺、(•)OH、O₂(•-)和少量H₂O₂,来氧化典型底物(3,3,5,5-四甲基联苯胺(TMB))并导致颜色变化。基于GOx触发原位形成磷酸根封端的硫化镉量子点的氧化酶模拟物的生长,我们开发了一种简单且放大的比色测定法来探测GOx的酶活性。该方法能够检测25μg/L至50mg/L范围内的GOx酶活性,检测限低至6.6μg/L。我们相信,原位生成的具有光刺激酶模拟活性的磷酸根封端的硫化镉量子点可能在生物传感器中具有广泛的潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/499b/6273347/1d4db071bbd5/molecules-21-00902-g001.jpg

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