He Jianwei, Zhang Xiaoyan, Qian Yuanyi, Wang Qiyao, Bai Yunpeng
State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing, East China University of Science and Technology, Shanghai, 200237, PR China.
State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing, East China University of Science and Technology, Shanghai, 200237, PR China.
Biosens Bioelectron. 2022 Jun 15;206:114085. doi: 10.1016/j.bios.2022.114085. Epub 2022 Feb 17.
The environmental accumulation of organophosphates is a serious threat to public health. To detect these xenobiotics, a broad range of sensors has been developed in past decades. However, sensors with high sensitivity and a capability for degrading organophosphates are rare. In this study, "smart" whole-cell biosensors were created by combining synthetic biology approaches with the bacterial quorum sensing (QS) mechanism. The engineered whole-cell biosensor pUC57-QS-DSF-F42 L/E coli DH5α can sense a wide array of phenolic compounds including phenol and p-nitrophenol (p-NP). By optimizing the genetic circuits, the phenol and p-NP detection limits reached 0.1 and 1 μM, respectively. Importantly, by replacing the fluorescence-generated reporter sfGFP with MP-degrading enzyme PoOPH, the whole-cell biosensor pUC57-OPH-QS-DSF-F42 L/E coli DH5α actively degraded 10 and 100 μM methyl parathion (MP), a typical organophosphate pesticide, which was artificially added to the cell culture at different time points in five consecutive degrading experiments, demonstrating its MP sensing and degrading capabilities. The universal design of this new biosensor can be used to create more efficient biosensors to detect and degrade various pollutants in the environment for rapid testing and bioremediation.
有机磷酸盐在环境中的积累对公众健康构成严重威胁。为了检测这些外源性物质,在过去几十年中已经开发了各种各样的传感器。然而,具有高灵敏度且能够降解有机磷酸盐的传感器却很少见。在本研究中,通过将合成生物学方法与细菌群体感应(QS)机制相结合,创建了“智能”全细胞生物传感器。工程化的全细胞生物传感器pUC57-QS-DSF-F42 L/E coli DH5α能够检测包括苯酚和对硝基苯酚(p-NP)在内的多种酚类化合物。通过优化遗传电路,苯酚和对硝基苯酚的检测限分别达到了0.1和1 μM。重要的是,通过用MP降解酶PoOPH取代产生荧光的报告基因sfGFP,全细胞生物传感器pUC57-OPH-QS-DSF-F42 L/E coli DH5α在连续五次降解实验的不同时间点被人工添加到细胞培养物中的典型有机磷酸酯农药甲基对硫磷(MP),分别为10和100 μM时,能够主动降解,证明了其对MP的传感和降解能力。这种新型生物传感器的通用设计可用于创建更高效的生物传感器,以检测和降解环境中的各种污染物,用于快速检测和生物修复。