酶级联反应放大策略用于灵敏检测酸性磷酸酶

引用本文: 李苹, 董玉明, 王光丽. 酶级联反应放大策略用于灵敏检测酸性磷酸酶[J]. 分析化学, 2018, 46(7): 1129-1136. doi: 10.11895/j.issn.0253-3820.181038 shu

Citation: LI Ping, DONG Yu-Ming, WANG Guang-Li. An Enzyme Cascade Strategy for Sensitive Detection of Acid Phosphatase[J]. Chinese Journal of Analytical Chemistry, 2018, 46(7): 1129-1136. doi: 10.11895/j.issn.0253-3820.181038 shu

酶级联反应放大策略用于灵敏检测酸性磷酸酶

江南大学合成与生物胶体教育部重点实验室化学与材料工程学院, 无锡 214112
基金项目:
本文系国家自然科学基金项目（Nos.21275065，21676123，21575052）资助

摘要: 构建了一个新型酶级联反应体系用于灵敏检测酸性磷酸酶（ACP）。焦磷酸根离子（PPi）与Fe³⁺有很强的结合作用，阻碍了具有模拟过氧化物酶活性的普鲁士蓝纳米粒子（PBNPs）的生成。ACP可将底物PPi水解为PO₄^3-，从而释放出Fe³⁺。释放的Fe³⁺与亚铁氰化钾（K₄[Fe（CN）₆]）形成PBNPs，PBNPs可以催化过氧化氢（H₂O₂）产生羟基自由基（·OH），进而催化氧化特征底物3，3'，5，5'-四甲基联苯胺（TMB），使体系产生明显的颜色变化，同时伴随吸光度的变化。基于上述原理，构建了一种灵敏检测ACP的方法。结果表明，在3~20 U/L的浓度范围内，氧化TMB（oxTMB）的吸光度与ACP浓度呈较好的线性关系，检出限为0.8 U/L。本研究中，通过天然酶的催化反应产生纳米材料模拟酶构建的酶级联催化反应体系具有高效的信号放大作用，有望在生物传感领域发挥重要作用。

关键词:

English

An Enzyme Cascade Strategy for Sensitive Detection of Acid Phosphatase

Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, China
Received Date: 17 January 2018
Revised Date: 20 April 2018
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 21275065, 21676123, 21575052).

Abstract: An enzyme cascade strategy was introduced for sensitive detection of acid phosphatase (ACP). Pyrophosphate ions (PPi) can strongly bound Fe³⁺ and thus hinders the production of Prussian blue nanoparticles (PBNPs). ACP can hydrolyze PPi to form phosphate ions, and the released Fe³⁺ reacts with potassium ferrocyanide (K₄[Fe(CN)₆]) to form PBNPs. The formed PBNPs have high peroxidase-like activity, which can decompose hydrogen peroxide (H₂O₂) to produce hydroxyl radical (·OH) for oxidizing the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB). Therefore, a novel sensing strategy for detecting ACP based on the high signal amplification of enzyme cascade was constructed. The results showed that there was a good linear relationship between the absorbance of oxidized TMB (oxTMB) and the concentration of ACP in the range of 3-20 U/L, with a detection limit of 0.8 U/L. Different from the conventional enzyme cascades in which the product of one enzyme is the substrate of the other, this study opens up a new way to construct novel enzyme cascade system.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

酶级联反应放大策略用于灵敏检测酸性磷酸酶

English

An Enzyme Cascade Strategy for Sensitive Detection of Acid Phosphatase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

酶级联反应放大策略用于灵敏检测酸性磷酸酶

English

An Enzyme Cascade Strategy for Sensitive Detection of Acid Phosphatase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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