Protonation-induced DNA conformational-change dominated electrochemical platform for glucose oxidase and urease analysis

Dandan Hu; Di Wu; Yanmei Lu; Jiayue Liu; Zhiyong Guo; Sui Wang; Chunyang Zhai; Zhihe Qing; Yufang Hu

doi:10.1016/j.aca.2022.340164

Protonation-induced DNA conformational-change dominated electrochemical platform for glucose oxidase and urease analysis

Anal Chim Acta. 2022 Sep 15:1226:340164. doi: 10.1016/j.aca.2022.340164. Epub 2022 Jul 21.

Authors

Dandan Hu¹, Di Wu², Yanmei Lu³, Jiayue Liu³, Zhiyong Guo³, Sui Wang³, Chunyang Zhai³, Zhihe Qing⁴, Yufang Hu⁵

Affiliations

¹ State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, State Key Laboratory Base of Novel Functional Materials and Preparation Science, School of Materials Science & Chemical Engineering, Ningbo University, Ningbo, Zhejiang, 315211, China; Hunan Provincial Key Laboratory of Cytochemistry, School of Chemistry and Chemical Engineering, Changsha University of Science and Technology, Changsha, 410114, China.
² College of Medical Technology, Ningbo College of Health Sciences, Ningbo, Zhejiang, 315100, China. Electronic address: wudi8412@163.com.
³ State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, State Key Laboratory Base of Novel Functional Materials and Preparation Science, School of Materials Science & Chemical Engineering, Ningbo University, Ningbo, Zhejiang, 315211, China.
⁴ Hunan Provincial Key Laboratory of Cytochemistry, School of Chemistry and Chemical Engineering, Changsha University of Science and Technology, Changsha, 410114, China.
⁵ State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, State Key Laboratory Base of Novel Functional Materials and Preparation Science, School of Materials Science & Chemical Engineering, Ningbo University, Ningbo, Zhejiang, 315211, China; Hunan Provincial Key Laboratory of Cytochemistry, School of Chemistry and Chemical Engineering, Changsha University of Science and Technology, Changsha, 410114, China. Electronic address: huyufang@nbu.edu.cn.

PMID: 36068049
DOI: 10.1016/j.aca.2022.340164

Abstract

Cytosine and protonated cytosine base pairs (C·CH⁺)-supported i-motif conformation has been widely employed in some interdisciplinary fields such as biology, medicine and chemistry. In this work, we report a new electrochemical biosensing method for the detection of glucose oxidase (GOx) and urease based on pH-induced DNA conformational-change. The constructed platform mainly includes TdT-mediated catalytic synthesis, GOx- or urease-catalyzed biological reaction and pH-induced DNA conformational-change. In the beginning, a kind of C-rich DNA is produced by TdT catalysis, and multiple C·CH⁺-supported i-motif structures appear under acidic condition. Then, the oxidation of glucose catalyzed by GOx or the hydrolyzation of urea aroused by urease can result in a generation of acidic or alkaline environment owing to the generated gluconic acid or ammonia. Herein, protonation and deprotonation interaction in TdT-yielded C-rich DNA can lead to different electrochemical impedance spectroscopy (EIS) toward Fe(CN)₆^3-/4-. Based on it, the EIS response changes proportionally toward GOx concentrations from 0.01 to 20 U/L or urease concentrations from 0.01 to 50 U/L, and the detection limit of GOx or urease is 0.0061 U/L or 0.0028 U/L (S/N = 3), respectively. Beyond this, we also construct a series of molecular logic gates (YES, AND, NOT, and NAND) with good performance by altering inputs under long C-rich DNA substrate. These excellent properties indicate that the unique sensing platform is potential to monitor GOx or urease in practical biosystems and clinical medical examinations.

Keywords: DNA conformational-Change; EIS platform; Glucose oxidase; I-motifs; TdT catalysis; Urease.

MeSH terms

Biosensing Techniques* / methods
Cytosine
Electrochemical Techniques / methods
Glucose / analysis
Glucose Oxidase* / chemistry
Urease

Substances

Cytosine
Glucose Oxidase
Urease
Glucose