Effects of Nano Zero-Valent Iron on Klebsiella oxytoca and Stress Response

Nano zero-valent iron (NZVI) is a new option for contaminated soil and groundwater treatment, despite little is known on their impact on environmental microorganisms. Klebsiella oxytoca K5 strain, isolated from the NZVI-treated soil, was used to investigate the bacterial, phenotypical and molecular response to commercial NZVI exposure. Cytotoxicity assays at three NZVI concentrations (1, 5 and 10 mg mL(-1)) suggested a negligible bacteriostatic effect and the lack of bactericidal effect. Structural changes were analysed by electronic microscopy. Scanning electron microscopy revealed the presence of NZVI around some bacterial cells, but no apparent morphological changes were seen. NZVI attachment to the cell surface was confirmed by transmission electron microscopy, although most of them were not affected. A proteomic approach (two-dimensional electrophoresis, matrix-assisted laser desorption ionization time-of-flight mass spectrometry) was used to investigate NZVI impact. For the first time to our knowledge, results revealed that exposure of a soil bacterium to NZVI resulted in the overproduction of tryptophanase, associated with oxidative stress response. K5 may set up an adaptative stress response involving indole as a signal molecule to inform the bacterial population about environmental changes. These findings would improve knowledge on the molecular mechanisms underlying bacterial response to NZVI exposure.