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纳米酶和天然酶的生物偶联以实现一步级联反应,用于代谢物的检测。

Bioconjugation of nanozyme and natural enzyme to enable a one-step cascade reaction for the detection of metabolites.

机构信息

Tianjin Key Laboratory On Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Building B for School of Pharmacy, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin, 300072, China.

出版信息

Anal Bioanal Chem. 2023 Jul;415(17):3385-3398. doi: 10.1007/s00216-023-04720-9. Epub 2023 May 4.


DOI:10.1007/s00216-023-04720-9
PMID:37140675
Abstract

Nanozyme, with enzyme-mimicking activity and excellent stability, has attracted extensive attention. However, some inherent disadvantages, including poor dispersion, low selectivity, and insufficient peroxidase-like activity, still limit its further development. Therefore, an innovative bioconjugation of a nanozyme and natural enzyme was conducted. In the presence of graphene oxide (GO), histidine magnetic nanoparticles (H-FeO) were first synthesized by a solvothermal method. The GO-supported H-FeO (GO@H-FeO) exhibited superior dispersity and biocompatibility because GO was the carrier and possessed outstanding peroxidase-like activity because of the introduction of histidine. Furthermore, the mechanism of the peroxidase-like activity of GO@H-FeO was the generation of •OH. Uric acid oxidase (UAO) was selected as the model natural enzyme and covalently linked to GO@H-FeO with hydrophilic poly(ethylene glycol) as a linker. UAO could specifically catalyze the oxidation of uric acid (UA) to generate HO, and subsequently, the newly produced HO oxidized the colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue ox-TMB under the catalysis of GO@H-FeO. Based on the above cascade reaction, the GO@H-FeO-linked UAO (GHFU) and GO@H-FeO-linked ChOx (GHFC) were used for the detection of UA in serum samples and cholesterol (CS) in milk, respectively. The method based on GHFU exhibited a wide detection range (5-800 μM) and a low detection limit (1.5 μM) for UA, and the method based on GHFC exhibited a wide detection range (4-400 μM) and a low detection limit (1.13 μM) for CS. These results demonstrated that the proposed strategy had great potential in the field of clinical detection and food safety.

摘要

纳米酶具有模拟酶的活性和优异的稳定性,引起了广泛关注。然而,一些固有缺点,包括较差的分散性、低选择性和不足的过氧化物酶样活性,仍然限制了其进一步发展。因此,进行了纳米酶与天然酶的创新生物缀合。在氧化石墨烯(GO)存在的情况下,通过溶剂热法首先合成了组氨酸磁性纳米粒子(H-FeO)。GO 负载的 H-FeO(GO@H-FeO)表现出优异的分散性和生物相容性,因为 GO 是载体,并由于引入组氨酸而具有出色的过氧化物酶样活性。此外,GO@H-FeO 的过氧化物酶样活性的机制是生成•OH。尿酸氧化酶(UAO)被选为模型天然酶,并通过亲水性聚乙二醇(PEG)作为连接子共价连接到 GO@H-FeO。UAO 可以特异性催化尿酸(UA)的氧化生成 HO,随后,新生成的 HO 在 GO@H-FeO 的催化下氧化无色 3,3',5,5'-四甲基联苯胺(TMB)为蓝色 ox-TMB。基于上述级联反应,用于检测血清样品中 UA 和牛奶中胆固醇(CS)的 GO@H-FeO 连接的 UAO(GHFU)和 GO@H-FeO 连接的 ChOx(GHFC)被分别用于检测 UA 和 CS。基于 GHFU 的方法对 UA 的检测范围很宽(5-800 μM),检测限很低(1.5 μM),基于 GHFC 的方法对 CS 的检测范围很宽(4-400 μM),检测限很低(1.13 μM)。这些结果表明,所提出的策略在临床检测和食品安全领域具有巨大的潜力。

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本文引用的文献

[1]
Depletable peroxidase-like activity of FeO nanozymes accompanied with separate migration of electrons and iron ions.

Nat Commun. 2022-9-12

[2]
Development of a Nonenzymatic Colorimetric Sensor for the Detection of Uric Acid Based on Ionic Liquid-Mediated Nickel Nanostructures.

ACS Omega. 2022-7-20

[3]
A colorimetric nanoprobe based on enzyme-immobilized silver nanoparticles for the efficient detection of cholesterol.

RSC Adv. 2019-12-18

[4]
Multienzyme Cascades Based on Highly Efficient Metal-Nitrogen-Carbon Nanozymes for Construction of Versatile Bioassays.

Anal Chem. 2022-3-1

[5]
Magnetic ZIF-8-Based Mimic Multi-enzyme System as a Colorimetric Biosensor for Detection of Aryloxyphenoxypropionate Herbicides.

ACS Appl Mater Interfaces. 2021-9-22

[6]
Porous Au@Pt nanoparticles with superior peroxidase-like activity for colorimetric detection of spike protein of SARS-CoV-2.

J Colloid Interface Sci. 2021-12-15

[7]
Highly permselective uric acid detection using kidney cell membrane-functionalized enzymatic biosensors.

Biosens Bioelectron. 2021-10-15

[8]
Flexible electrochemical uric acid and glucose biosensor.

Bioelectrochemistry. 2021-10

[9]
A Multi-Enzyme Cascade Reaction for the Production of 2'3'-cGAMP.

Biomolecules. 2021-4-16

[10]
Pentacyanoammineferrate-Based Non-Enzymatic Electrochemical Biosensing Platform for Selective Uric Acid Measurement.

Sensors (Basel). 2021-2-24

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