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自组装的、血红素功能化的肽纳米管:一种具有过氧化物酶样活性检测谷胱甘肽和葡萄糖分子的创新策略。

Self-assembled, hemin-functionalized peptide nanotubes: an innovative strategy for detecting glutathione and glucose molecules with peroxidase-like activity.

作者信息

Xiang Song, Long Xincheng, Tu Qiuxia, Feng Jian, Zhang Xiaohe, Feng Guangwei, Lei Li

机构信息

Key Laboratory of Microbiology and Parasitology of Education, Department of Guizhou, School of Basic Medical Science, Guizhou Medical University, Guiyang, China.

School of Clinical Laboratory Science, Guizhou Medical University, Guiyang, 550025, China.

出版信息

Nano Converg. 2023 Feb 4;10(1):7. doi: 10.1186/s40580-023-00356-8.

Abstract

Accurately detecting dynamic changes in bioactive small molecules in real-time is very challenging. In this study, a hemin-based peptide assembly was rationally designed for the colorimetric detection of active small molecules. Hemin-functionalized peptide nanotubes were obtained through the direct incubation of hemin (hemin@PNTs) and peptide nanotubes (PNTs) or were coassembled with the heptapeptide Ac-KLVFFAL-NH via electrostatic, π-π stacking, and hydrophobic interactions (hemin-PNTs). This new substance is significant because it exhibits the benefits of both hemin and PNTs as well as some special qualities. First, hemin-PNTs exhibited higher intrinsic peroxidase-like activity, which, in the presence of HO, could catalyze the oxidation of the substrate 3,3',5,5'-tetramethylbenzidine (TMB) to yield a typical blue solution after 10 min at 25 ℃. Second, hemin-PNTs showed significantly higher activity than that of hemin, PNTs alone, or hemin@PNTs. Hemin-PNTs with a 20.0% hemin content may cooperate to improve catalytic activity. The catalytic activity was dependent on the reaction temperature, pH, reaction time, and HO concentration. The nature of the TMB-catalyzed reaction may arise from the production of hydroxyl radicals. Fluorescence analysis was used to demonstrate the catalytic mechanism. According to this investigation, a new highly selective and sensitive colorimetric technique for detecting glutathione (GSH), L-cysteine, and glucose was established. The strategy demonstrated excellent sensitivity for GSH in the range of 1 to 30 μM with a 0.51 μM detection limit. Importantly, this glucose detection technique, which employs glucose oxidase and hemin-PNTs, is simple and inexpensive, with a 0.1 μM to 1.0 mM linear range and a 15.2 μM detection limit. Because of their low cost and high catalytic activity, hemin-PNTs are an excellent choice for biocatalysts in a diverse range of potential applications, including applications in clinical diagnostics, environmental chemistry, and biotechnology.

摘要

实时准确检测生物活性小分子的动态变化极具挑战性。在本研究中,合理设计了一种基于血红素的肽组装体用于比色检测活性小分子。通过血红素(hemin@PNTs)与肽纳米管(PNTs)直接孵育获得血红素功能化肽纳米管,或通过静电、π-π堆积和疏水相互作用与七肽Ac-KLVFFAL-NH共组装(hemin-PNTs)。这种新物质具有重要意义,因为它兼具血红素和PNTs的优点以及一些特殊性质。首先,hemin-PNTs表现出更高的内在过氧化物酶样活性,在HO存在下,25℃反应10分钟后,可催化底物3,3',5,5'-四甲基联苯胺(TMB)氧化生成典型的蓝色溶液。其次,hemin-PNTs的活性显著高于单独的血红素、PNTs或hemin@PNTs。血红素含量为20.0%的hemin-PNTs可能协同提高催化活性。催化活性取决于反应温度、pH值、反应时间和HO浓度。TMB催化反应的本质可能源于羟基自由基的产生。采用荧光分析来证明催化机制。根据本研究,建立了一种用于检测谷胱甘肽(GSH)、L-半胱氨酸和葡萄糖的新型高选择性和灵敏比色技术。该策略对1至30μM范围内的GSH表现出优异的灵敏度,检测限为0.51μM。重要的是,这种采用葡萄糖氧化酶和hemin-PNTs的葡萄糖检测技术简单且成本低廉,线性范围为0.1μM至1.0mM,检测限为15.2μM。由于成本低且催化活性高,hemin-PNTs是多种潜在应用中生物催化剂的极佳选择,包括临床诊断、环境化学和生物技术应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c543/9899300/05711ee79ba9/40580_2023_356_Fig1_HTML.jpg

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