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通过物理生物控制和两级信号放大构建稳定且灵敏的工程细菌传感器。

A Stable and Sensitive Engineering Bacterial Sensor via Physical Biocontainment and Two-Stage Signal Amplification.

机构信息

Faculty of Materials Science and Chemistry, China University of Geosciences, Wuhan 430074, China.

Civil and Environmental Engineering, University of Hawai'i, Honolulu Hawai'i 96822, United States.

出版信息

Anal Chem. 2024 May 28;96(21):8807-8813. doi: 10.1021/acs.analchem.4c01341. Epub 2024 May 7.

Abstract

Although engineering bacterial sensors have outstanding advantages in reflecting the actual bioavailability and continuous monitoring of pollutants, the potential escape risk of engineering microorganisms and lower detection sensitivity have always been one of the biggest challenges limiting their wider application. In this study, a core-shell hydrogel bead with functionalized silica as the core and alginate-polyacrylamide as the shell have been developed not only to realize zero escape of engineered bacteria but also to maintain cell activity in harsh environments, such as extremely acidic/alkaline pH, high salt concentration, and strong pressure. Particularly, after combining the selective preconcentration toward pollutants by functionalized core and the positive feedback signal amplification of engineering bacteria, biosensors have realized two-stage signal amplification, significantly improving the detection sensitivity and reducing the detection limit. In addition, this strategy was actually applied to the detection of As(III) and As(V) coexisting in environmental samples, and the detection sensitivity was increased by 3.23 and 4.39 times compared to sensors without signal amplification strategy, respectively, and the detection limits were as low as 0.39 and 0.86 ppb, respectively.

摘要

虽然工程细菌传感器在反映污染物的实际生物可利用性和连续监测方面具有突出的优势,但工程微生物的潜在逃逸风险和较低的检测灵敏度一直是限制其更广泛应用的最大挑战之一。在这项研究中,开发了一种具有功能化二氧化硅核和海藻酸钠-聚丙烯酰胺壳的核壳水凝胶珠,不仅实现了工程细菌的零逃逸,而且还保持了细胞在极端酸性/碱性 pH 值、高盐浓度和强压力等恶劣环境中的活性。特别是,通过功能化核心对污染物的选择性预浓缩和工程细菌的正反馈信号放大相结合,生物传感器实现了两级信号放大,显著提高了检测灵敏度,降低了检测限。此外,该策略实际上已应用于环境样品中 As(III)和 As(V)的同时检测,与没有信号放大策略的传感器相比,检测灵敏度分别提高了 3.23 倍和 4.39 倍,检测限分别低至 0.39 和 0.86 ppb。

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