An unexpected discovery of 1,4-benzoquinone as a lipophilic mediator for toxicity detection in water

Abstract

Since most toxicological risk assessments are based on individual single-species tests, there is uncertainty in extrapolating these results to ecosystem assessments. Herein, we successfully developed a mediated microbial electrochemical biosensor with mixed microorganisms for toxicity detection by microelectrode array (MEA). In order to fully mobilize all the mixed microorganisms to participate in electron transfer to amplify the current signal, 1,4-benzoquinone (BQ) was used as the lipophilic mediator to mediate the intracellular metabolic activities. Hydrophilic K₃[Fe(CN)₆] was employed as an extracellular electron acceptor to transport electrons from hydroquinone (HQ) to the working electrode. Under the optimal conditions of 50 mM phosphate buffer solution (PBS), 0.4 mM BQ, 10 mM K₃[Fe(CN)₆] and OD₆₀₀ = 0.5 bacteria concentration, the half-maximal inhibitory concentration (IC₅₀) values measured with the composite-mediated respiration (CM-RES) of BQ–K₃[Fe(CN)₆] for Cu²⁺, Cd²⁺ and Zn²⁺ were 5.95, 7.12 and 8.86 mg L⁻¹, respectively. IC₅₀ values obtained with the single mediator K₃[Fe(CN)₆] were 2.34, 5.88 and 2.42 mg L⁻¹ for the same samples. The results indicate that the biosensor with the single mediator K₃[Fe(CN)₆] had higher sensitivity to heavy metal ions than the biosensor with composite mediators. After verification, we found that the addition of BQ cannot amplify the current. The IC₅₀ value of 0.89 mg L⁻¹ for BQ was obtained using K₃[Fe(CN)₆] as the single mediator. This suggests that BQ is highly toxic, which explained why the sensitivity of the biosensor with the combined mediator BQ–K₃[Fe(CN)₆] was lower than that of the biosensor with the single mediator K₃[Fe(CN)₆]. At the same time, this also implies that toxicity itself cannot be ignored when it is used as an electronic mediator in a mediated microbial electrochemical biosensor.