Development of a stable luciferase@nanoflower-based method for the sensitive detection of pesticide residues in milk

IF 9.8 1区农林科学 Q1 CHEMISTRY, APPLIED

Food Chemistry Pub Date : 2025-04-14 DOI:10.1016/j.foodchem.2025.144323

Xiaoyu Li , Fengchun Huang , Juan Zhang , Jiaci Chen , Xiangyi Pang , Longrui Yang , Xiaoyun Sun , Yuhang Fan , Ailiang Chen , Qingli Yang

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Abstract

Acetylcholinesterase inhibition–luciferase bioluminescence system enables rapid detection of organophosphate and carbamate pesticide residues. However, firefly luciferase (FLuc) suffers from poor stability and short luminescence duration, thereby introducing uncertainties in detection. To address this problem, nanoflower-immobilized FLuc (FLuc@NFs) was prepared using a coprecipitation method, which markedly enhanced stability and activity. FLuc@NFs exhibited intact nanoflower structures and allowed storage at 25 °C and at 4 °C for 2 weeks, overcoming the limitation of free enzyme storage at −20 °C. In the bioluminescence reaction, FLuc@NFs at 1 mg/mL achieved 62.14 % higher luminescence than free FLuc, with stable signals extending from 10 s to 30 min, enabling high-throughput detection. The system achieved detection limits of 5 and 10 ng/mL for chlorpyrifos and carbaryl, respectively, demonstrating improved reliability and sensitivity in pesticide detection.

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基于稳定荧光素酶@纳米花的牛奶中农药残留灵敏检测方法的开发

乙酰胆碱酯酶抑制-荧光素酶生物发光系统实现了有机磷和氨基甲酸酯类农药残留的快速检测。然而，萤火虫荧光素酶（FLuc）稳定性差，发光时间短，因此在检测中存在不确定性。为了解决这一问题，采用共沉淀法制备了纳米花固定化FLuc (FLuc@NFs)，其稳定性和活性显著提高。FLuc@NFs表现出完整的纳米花结构，并允许在25 °C和4 °C下储存2 周，克服了在- 20 °C下自由酶储存的限制。在生物发光反应中，FLuc@NFs在1 mg/mL浓度下的发光比游离FLuc高62.14 %，稳定信号从10 s延伸到30 min，实现了高通量检测。该系统对毒死蜱和威威的检出限分别为5 ng/mL和10 ng/mL，提高了农药检测的可靠性和灵敏度。

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来源期刊

Food Chemistry 工程技术-食品科技

CiteScore

16.30

自引率

10.20%

发文量

3130

审稿时长

122 days

期刊介绍： Food Chemistry publishes original research papers dealing with the advancement of the chemistry and biochemistry of foods or the analytical methods/ approach used. All papers should focus on the novelty of the research carried out.