Mechanistic evaluation of enhanced graphene toxicity to Bacillus induced by humic acid adsorption

The extensive application of graphene nanosheets (GNSs) has raised concerns over risks to sensitive species in the aquatic environment. The humic acid (HA) corona is traditionally considered to reduce GNSs toxicity. Here, we evaluate the effect of sorbed HA (GNSs-HA) on the toxicity of GNSs to Gram positive Bacillus tropicus. Contrary to previous data, GNSs-HA exhibits greater toxicity compared to GNSs. Multi-omics combined with sensitive bioassays and electrochemical methods reveals GNSs disrupt oxidative phosphorylation by causing physical membrane damage. This leads to the accumulation of intracellular reactive oxygen species and inhibition of ATP production, subsequently suppressing synthetic and metabolic processes and ultimately causing bacterial death. Conversely, GNSs-HA directly extracts electrons from bacteria and oxidized biomolecules due to HA-improved electron transfer. This finding suggests that the HA corona does not always mitigate the toxicity of nanoparticles, thereby introducing uncertainty over the interaction between environmental corona and nanoparticles during ecological risk evaluation.