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通过光诱导电子转移(PET)过程对漆酶活性进行荧光检测。

Fluorescence detection of laccases activity by the photoinduced electron transfer (PET) process.

机构信息

Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgān, Iran.

Nano Drug Delivery Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.

出版信息

J Biol Inorg Chem. 2020 Feb;25(1):151-159. doi: 10.1007/s00775-019-01748-0. Epub 2019 Dec 12.

DOI:10.1007/s00775-019-01748-0
PMID:31832782
Abstract

Laccases play a vital role in some physiological processes, for example in morphogenesis, carbon cycle, and defense against parasitism. So, designing a high-sensitivity accurate method is essential for researchers. In this study, a simple fluorescence method based on the function of carbon nitride (g-CN) by dopamine is synthesized. For the design of this sensor, carbon nitride (g-CN) is initially synthesis by using a simple method, which is carried out by heating melamine at 550 °C for 3 h and modifying it with dopamine by a linker such as glutaraldehyde. However, the g-CN-Dopa produced by this method, with an excitation wavelength of 330 nm, has a fluorescence emission at 466 nm. When laccase and g-CN-Dopa were mixed, dopamine with redox property was oxidized to dopaquinone; this causes the phenomenon of photoinduced electron transfer (PET) process between g-CN and the dopaquinone. Hence, fluorescence quenching occurs due to this phenomenon. As a result of these discussions, a sensor for the laccase activity was designed based on the fluorescence quenching degree, supporting a linear range of 0.0-400.0 U L with the detection limit of 2.0 U L. Using this sensor, the activity of the laccase enzyme in the human serum samples is measured. Dopamine-functionalized carbon nitride was prepared and utilized for the highly sensitive detection of laccases activity.

摘要

漆酶在一些生理过程中起着至关重要的作用,例如形态发生、碳循环和抵御寄生虫。因此,设计一种高灵敏度、准确的方法对研究人员来说至关重要。在这项研究中,设计了一种基于多巴胺功能的氮化碳(g-CN)的简单荧光方法。为了设计这个传感器,首先通过在 550°C 下加热三聚氰胺 3 小时的简单方法合成氮化碳(g-CN),并用戊二醛等连接子对其进行多巴胺修饰。然而,用这种方法制备的 g-CN-Dopa 在 330nm 的激发波长下,在 466nm 处有荧光发射。当漆酶和 g-CN-Dopa 混合时,具有氧化还原性质的多巴胺被氧化成多巴醌;这导致 g-CN 和多巴醌之间发生光诱导电子转移(PET)过程。因此,由于这种现象发生了荧光猝灭。基于这些讨论,设计了一种基于荧光猝灭程度的漆酶活性传感器,其线性范围为 0.0-400.0 U L,检测限为 2.0 U L。使用这个传感器,测量了人血清样品中漆酶酶的活性。制备了多巴胺功能化的氮化碳,并将其用于高灵敏度检测漆酶的活性。

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