Humic acids facilitated microbial reduction of polymeric Pu(IV) under anaerobic conditions

Flavins and humic substances have been extensively studied with emphasis on their ability to transfer extracellular electrons to insoluble metal oxides. Nevertheless, whether the low-solubility Pu(IV) polymers are microbially reduced to aqueous Pu(III) remains uncertain. Experiments were conducted under anaerobic and slightly alkaline conditions to study the difference between humic acids and flavins to transport extracellular electrons to Pu(IV) polymers. Our study demonstrates that Shewanella putrefaciens was unable to directly reduce polymeric Pu(IV) with a notably low reduction rate (3.4×10^-12mol/L Pu(III)_aq within 144h). The relatively high redox potential of flavins reveals the thermodynamically unfavorable reduction: E_h(PuO₂(am)/Pu³⁺)h^o'(FMN/FMNH₂)≈E_h^o'(RBF/RBFH₂)≈-220mV at pH7.2. The microbially reduced humic acids facilitated the extracellular electron transfer to the polymers and reduced polymeric Pu(IV) (2.1×10^-10mol/L Pu(III)_aq) 62 times more rapidly than the flavins. The driving force for electron transfer explains the observed reduction: E_h(HA_ox/HA_red)h(PuO₂(am)/Pu³⁺) when S. putrefaciens oxidized lactate and respired on the humic acids. In contrast, flavins were able to substantially reduce aqueous Pu(IV)-EDTA (1.9×10^-9mol/L Pu(III)_aq) because of the available driving force for electron transfer: Δ_rG_m=-F[E_h(PuL₂^4-/PuL₂^5-)-E_h^o'(FMN/FMNH₂)]=-33.5kJ/mol is a result of E_h(PuL₂^4-/PuL₂^5-)≫E_h(PuO₂(am)/Pu³⁺), where L is the EDTA ligand. In the presence of humic acids, the reduction of Pu(IV)-EDTA exhibited the most rapid rate (2.2×10^-9mol/L Pu(III)_aq). This result further demonstrates that humic acids facilitated the extracellular electron transfer to polymeric and aqueous Pu(IV). Reductive solubilization of the polymers may enhance Pu mobility in the geosphere and hence increases risks to human health.