Fast preparing bioelectrode with conductive bioink for nitrite detection in high sensitivity and stability

Electrochemically active biofilms (EABs) for nitrite detection have high specificity, rapid response, operational simplicity, and extended lifespan advantages. However, their scale production remains challenging due to time-consuming and uniform preparation. In this study, a novel approach was proposed to fast fabricate an EAB biosensor with a synthetic biofilm electrode for nitrite detection. The biofilm electrode was prepared by coating bioinks with varying conductive materials onto the surface of the graphite sheets, showing short incubation time and good reproducibility. Incorporating conductive materials into the bioinks remarkably enhanced the maximum voltage of the first cycle of bioelectrode incubation, with an increase of up to 633% for carbon nanofibers. The nitrite reduction current was amplified by a factor of 2.97, due to the enhancement of extracellular electron transfer (EET). The developed nitrite biosensor exhibited a detection range of 0.1-15 mg NO₂^--N L^-1, with a high sensitivity of 610.8 μA mM^-1 cm^-2, and a stabilization operation time of at least 280 cycles. This study not only provided valuable insights into conductive materials for synthetic biofilms but also presented a practical approach for the rapid preparation, scale production, and optimization of highly sensitive and stable EAB sensors.