A copper-specific microbial fuel cell biosensor based on riboflavin biosynthesis of engineered Escherichia coli.

Copper pollution poses a serious threat to the aquatic environment; however, in situ analytical methods for copper monitoring are still scarce. In the current study, Escherichia coli Rosetta was genetically modified to express OprF and ribB with promoter Pt7 and PcusC , respectively, which could synthesize porin and senses Cu2+ to produce riboflavin. The cell membrane permeability of this engineered strain was increased and its riboflavin production (1.45-3.56 µM) was positively correlated to Cu2+ (0-0.5 mM). The biosynthetic strain was then employed in microbial fuel cell (MFC) based biosensor. Under optimal operating parameters of pH 7.1 and 37°C, the maximum voltage (248, 295, 333, 352, and 407 mV) of the constructed MFC biosensor showed a linear correlation with Cu2+ concentration (0.1, 0.2, 0.3, 0.4, 0.5 mM, respectively; R2 = 0.977). The continuous mode testing demonstrated that the MFC biosensor specifically senses Cu2+ with calculated detection limit of 28 µM, which conforms to the common Cu2+ safety standard (32 µM). The results obtained with the developed biosensor system were consistent with the existing analytical methods such as colorimetry, flame atomic absorption spectrometry, and inductively coupled plasma optical emission spectrometry. In conclusion, this MFC-based biosensor overcomes the signal conversion and transmission problems of conventional approaches, providing a fast and economic analytical alternative for in situ monitoring of Cu2+ in water.

A Novel Early Warning System Based on a Sediment Microbial Fuel Cell for In Situ and Real Time Hexavalent Chromium Detection in Industrial Wastewater.

Hexavalent chromium (Cr(VI)) is a well-known toxic heavy metal in industrial wastewater, but in situ and real time monitoring cannot be achieved by current methods used during industrial wastewater treatment processes. In this study, a Sediment Microbial Fuel Cell (SMFC) was used as a biosensor for in situ real-time monitoring of Cr(VI), which was the organic substrate is oxidized in the anode and Cr(VI) is reduced at the cathode simultaneously. The pH 6.4 and temperature 25 °C were optimal conditions for the operation. Under the optimal conditions, linearity (R² = 0.9935) of the generated voltage was observed in the Cr(VI) concentration range from 0.2 to 0.7 mg/L. The system showed high specificity for Cr(VI), as other co-existing ions such as Cu2+, Zn2+, and Pb2+ did not interfere with Cr(VI) detection. In addition, when the sediment MFC-based biosensor was applied for measuring Cr(VI) in actual wastewater samples, a low deviation (<8%) was obtained, which indicated its potential as a reliable biosensor device. MiSeq sequencing results showed that electrochemically active bacteria (Geobacter and Pseudomonas) were enriched at least two-fold on the biofilm of the anode in the biosensor as compared to the SMFC without Cr(VI). Cyclic voltammetry curves indicated that a pair of oxidation/reduction peaks appeared at -111 mV and 581 mV, respectively. These results demonstrated that the proposed sediment microbial fuel cell-based biosensor can be applied as an early warning device for real time in situ detection of Cr(VI) in industrial wastewaters.

A novel biosensor for p-nitrophenol based on an aerobic anode microbial fuel cell.

P-nitrophenol is one of the most common contaminants in chemical industrial wastewater, and in situ real-time monitoring of PNP cannot be achieved by conventional analytical techniques. Here, a two-chamber microbial fuel cell with an aerobic anode chamber was tested as a biosensor for in situ real-time monitoring of PNP. Pseudomonas monteilii LZU-3, which was used as the biological recognition element, can form a biofilm on the anode electrode using PNP as a sole substrate. The optimal operation parameters of the biosensor were as follows: external resistance 1000Ω, pH 7.8, temperature 30°C, and maximum PNP concentration 50mgL(-1). Under these conditions, the maximum voltages showed a linear relationship with PNP concentrations ranging from 15±5 to 44±4.5mgL(-1). Furthermore, we developed a novel portable device for in situ real-time monitoring of PNP. When the device was applied to measure PNP in wastewater containing various additional aromatic compounds and metal ions, the performance of the biosensor was not affected and the correlation between the maximum voltages and the PNP concentrations ranging from 9±4mgL(-1) to 36 ± 5mgL(-1) was conserved. The results demonstrated that the MFC biosensor provides a rapid and cost-efficient analytical method for real-time monitoring of toxic and recalcitrant pollutants in environmental samples.