Resuscitation-promoting factor enhances PFOA biodegradation and modulates cellular stress responses in two PFOA-degrading bacterial strains.

Perfluorooctanoic acid (PFOA), a highly persistent and toxic per- and polyfluoroalkyl substance (PFAS), poses significant environmental and health risks. Microbial degradation offers a promising remediation strategy but is limited by the scarcity and low efficiency of known degraders. Here, two PFOA-degrading strains, Alcaligenes sp. MT1 and Pseudomonas sp. MT3, were isolated and characterized. Whole-genome analysis identified multiple genes potentially involved in fluorinated compound transformation. Supplementation with resuscitation-promoting factor (Rpf) significantly enhanced PFOA degradation, with an optimal dosage of 1.5% (v/v), increasing removal efficiencies by 16.8% (MT1) and 18.7% (MT3) relative to controls. Rpf improved cellular performance by elevating antioxidant enzyme activities (peroxidase, superoxide dismutase, catalase) and Na⁺/K⁺-ATPase activity while reducing malondialdehyde levels, indicating enhanced oxidative stress tolerance and energy metabolism. Consistently, key genes associated with decarboxylation and dehalogenation pathways were markedly upregulated. LC-MS/MS analysis revealed that PFOA degradation involved sequential decarboxylation and defluorination, with Rpf affecting metabolite abundance but not the core degradation pathways. These findings demonstrate that Rpf-mediated activation effectively enhances microbial PFOA degradation and provides a promising strategy for PFAS bioremediation.