Citation: LIU Tao, LI Dan, LIANG Jie, WANG Xiu-Mei. A Fluorescence Biosensor for Lead Ion Detection Based on DNAzyme and Exonuclease Ⅲ[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(2): 248-254. doi: 10.19756/j.issn.0253-3820.191491 shu

A Fluorescence Biosensor for Lead Ion Detection Based on DNAzyme and Exonuclease Ⅲ

Fujian Provincial Key Laboratory of Ecology-Toxicological Effects&Control for Emerging Contaminants College of Environmental and Biological Engineering, Putian University, Putian 351100, China

Received Date: 11 August 2019
Revised Date: 10 December 2019

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 31801462), the Scientific Research Foundation of Fujian Provincial Education Department (No. JT180466) and the Scientific Research Foundation of Putian University (No. 2017015).

A fluorescence sensor for Pb²⁺ detection was developed on the basis of a DNAzyme cleavage and exonuclease Ⅲ assistant amplification strategy. In the presence of Pb²⁺, the DNAzyme hybridized with substrate strand and catalyzed the hydrolytic cleavage of the substrate strand, and then the DNAzyme released from the substrate strand and bound another substrate strand to trigger another cycle of hydrolytic cleavage. The DNAzymes were used as catalysts for amplified sensing through multiple turnover reactions. The substrate probe was cleavaged and broken to form a Y-shaped probe which could hybridize and open the molecular beacon, resulting in the increase of the fluorescence signal. At the same time, exonuclease Ⅲ catalytically digested the molecular beacon from 3'-end and released the Y-shaped substrate strand. The released Y-shaped substrate strand could directly hybridize with another molecular beacon to generate fluorescence signal, and thus was further recognized and cleaved by exonuclease Ⅲ from the second step of cyclic signal amplification. Accompanying with each cleavage toward molecular probe by exonuclease Ⅲ, the fluorescence signal was accumulated, which resulted in a cyclic amplification format for the fluorescence response toward Pb²⁺ detection. The fluorescence response was detected in a 200 μL reaction system that was incubated at 37℃ for 60 min. The linear range for detection of Pb²⁺ was 0.05-200 nmol/L with a detection limit of 0.01 nmol/L, and the recoveries of environmental water samples were 96.3%-108.3%. This method had many advantages such as simple operation, rapid detection, high selectivity and high sensitivity, and showed great application potential in Pb²⁺ detection.
- Lead ion,
- DNAzyme,
- Exonuclease Ⅲ,
- Fluorescence biosensor
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