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将酪氨酸酶固定在 Feo@Au 核壳纳米粒子上作为生物探针,用于检测多巴胺、苯酚和儿茶酚。

Immobilization of tyrosinase on Feo@Au core-shell nanoparticles as bio-probe for detection of dopamine, phenol and catechol.

机构信息

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

Department of Chemistry, Faculty of Sciences, Azad University of Kermanshah, Kermanshah, Iran.

出版信息

J Biol Inorg Chem. 2019 Oct;24(7):961-969. doi: 10.1007/s00775-019-01691-0. Epub 2019 Jul 29.

Abstract

An optical bio-probe based on the immobilized tyrosinase on the surface of FeO@Au was described for the detection of dopamine, phenol and catechol. The prepared bio-probe (FeO@Au@tyrosinase) was characterized by means such as TEM, SEM, VSM, DLS and TGA. In the presence of the bio-probe, the phenol, catechol and dopamine were converted to benzoquinone, o-quinone and dopaquinone, and the fluorescence spectra appeared at 308 nm, 329 nm and 336 nm with ex = 270 nm, respectively. However, by increasing the concentration of phenolic compounds in the bio-probe, the amount of products (benzoquinone, o-quinone and dopaquinone) was increased which was the reason for the increase in fluorescence intensity. Using this mechanism, a bio-probe was designed such that the intensity of the fluorescence spectra increased proportionally with the increase of the substrate concentrations after different time periods. The 0.003 mg/mL of tyrosinase was loaded on 1.65 mg/mL of the FeO@Au. The highest performance for a bio-probe was demonstrated at room temperature and pH 6.8. By investigating the characteristics of the response of the bio-probe to different phenolic compounds, it was found that the bio-probe had a linear response in the concentration range 5.0-75.0 µM, 10.0-100.0 µM for phenol and dopamine and 50.0-500.0 M for catechol. The Michaelis-Menten constant (K) of the bio-probe was calculated as 0.6 µM. Finally, the bio-probe seems to be stable and efficient even after about 2 months. A novel and easy method for the detection of dopamine, phenol and catechol by florescence that uses oxide capability to identify the phenolic compounds was introduced.

摘要

一种基于固定在 FeO@Au 表面的酪氨酸酶的光学生物探针被描述用于检测多巴胺、苯酚和儿茶酚。通过 TEM、SEM、VSM、DLS 和 TGA 等手段对制备的生物探针(FeO@Au@tyrosinase)进行了表征。在生物探针的存在下,苯酚、儿茶酚和多巴胺被转化为苯醌、邻苯醌和多巴醌,荧光光谱分别在 308nm、329nm 和 336nm 处出现,激发波长为 270nm。然而,随着生物探针中酚类化合物浓度的增加,产物(苯醌、邻苯醌和多巴醌)的量增加,这是荧光强度增加的原因。利用这一机制,设计了一种生物探针,使得在不同时间段后,荧光光谱的强度随着底物浓度的增加而呈比例增加。在 1.65mg/mL 的 FeO@Au 上负载了 0.003mg/mL 的酪氨酸酶。该生物探针在室温下和 pH6.8 时表现出最佳性能。通过研究生物探针对不同酚类化合物响应的特性,发现生物探针在 5.0-75.0µM、10.0-100.0µM 范围内对苯酚和多巴胺以及 50.0-500.0µM 范围内对儿茶酚有线性响应。生物探针的米氏常数(K)计算为 0.6µM。最后,即使经过大约 2 个月,生物探针似乎仍然稳定且高效。介绍了一种通过氧化物能力识别酚类化合物的新型、简单的荧光检测多巴胺、苯酚和儿茶酚的方法。

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