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This study presents a novel bioelectrochemical sensor using self-assembled hydrogel bioelectrodes with reduced graphene oxide and genetically engineered Shewanella species for simultaneous detection of nitrate and nitrite. The sensor achieves sensitive and quantitative responses to both nitrite and nitrate, with a high sensitivity and low limit of detection. The whole-cell biohydrogel based sensor also exhibits excellent anti-interference and long-term storability, making it suitable for plug-and-play applications.
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Bioelectrochemical sensors (BES) are promising to specifically detect nitrate or nitrite but never realize simultaneous detection in a single system. In this study, a novel biosensor using self-assembled hydrogel bioelectrodes with reduced graphene oxide (rGO) and genetically engineered electroactive Shewanella species was designed for the simultaneous detection of nitrate and nitrite. The highly conductive rGO rendered a drastically improved reverse electron transfer from the electrode to Shewanella , which achieved a sensitive and quantitative response to nitrite/nitrate. Meanwhile, the genetically engineered Shewanella enabled efficient differentiation of nitrate and nitrite detection in a single system (a sensitivity of 883.48 μA mM -1 cm -2 to nitrate with a limit of detection of 0.92 μM and a sensitivity of 888.48 μA mM -1 cm -2 to nitrite with a limit of detection of 0.72 μM). The whole-cell biohydrogel based BES also showed excellent anti-interference and long-term storability for plug-and-play application. Based on these properties, this work demonstrated the power of genetically engineered electroactive bacteria in rGO biohydrogel for dual-analyte detection and also provided a new strategy for developing high-performance whole-cell BES for practical environmental monitoring.