Intra/extracellular electron transfer for aerobic denitrification mediated by in-situ biosynthesis palladium nanoparticles.

The slow electron transfer rate is the bottleneck to the biological wastewater treatment process, and the nanoparticles (NPs) has been verified as a feasible strategy to improve the biological degradation efficiency by accelerating the electron transfer. Here, we employed the Gram-positive Bacillus megaterium Y-4, capable of synthetizing Pd(0), to investigate the intra/extracellular electron transfer (IET/EET) mechanisms mediated by NPs in aerobic denitrification for the first time. Kinetic and thermodynamic results showed that the bio-Pd(0) could significantly promote the removal of both nitrate and nitrite by improving affinity and decreasing activation energy. The enzymic activity and the respiration chain inhibition experiment indicated that the bio-Pd(0) could facilitate the nitrate biotic reduction by improving the Fe-S center activity and serving as parallel H carriers to replace coenzyme Q to selectively increase the electron flux toward nitrate in IET, while promoting the nitrite reduction by abiotic catalysis. Most importantly, the detection of DPV peak at -226~-287 mV proved that the one-electron EET via multiheme cytochrome-bound flavins also occurred in Gram-positive bacteria and enhanced in Pd-loaded cells. In addition, the remarkable increase of the formal charge in EPS indicated that the bio-Pd(0) could act as an electron shuttle to increase the redox site in EPS, eventually accelerating the electron hopping in long-distance electron transfer. Overall, this study expanded our understanding of the roles of bio-Pd(0) on the aerobic denitrification process and provided an insight into the IET/EET of Gram-positive strains.