Massimiliano Magro1, Luca Fasolato2, Emanuela Bonaiuto3, Nadia Andrea Andreani4, Davide Baratella5, Vittorino Corraducci6, Giovanni Miotto7, Barbara Cardazzo8, Fabio Vianello9. 1. Department of Comparative Biomedicine and Food Science, University of Padova, Italy; Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Palacky University in Olomouc, Czech Republic. Electronic address: massimiliano.magro@unipd.it. 2. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: luca.fasolato@unipd.it. 3. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: emanuela.bonaiuto@unidp.it. 4. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: nadiaandrea.andreani@studenti.unipd.it. 5. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: davide.baratella@studenti.unipd.it. 6. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: vittorino.corraducci@unipd.it. 7. Department of Molecular Medicine, University of Padova, Italy. Electronic address: gianni.miotto@unipd.it. 8. Department of Comparative Biomedicine and Food Science, University of Padova, Italy. Electronic address: barbara.cardazzo@unipd.it. 9. Department of Comparative Biomedicine and Food Science, University of Padova, Italy; Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Palacky University in Olomouc, Czech Republic. Electronic address: fabio.vianello@unipd.it.
Abstract
BACKGROUND: Mineral iron(III) recognition by bacteria is considered a matter of debate. The peculiar surface chemistry of novel naked magnetic nanoparticles, called SAMNs (surface active maghemite nanoparticles) characterized by solvent exposed Fe(3+) sites on their surface, was exploited for studying mineral iron sensing in Pseudomonas fluorescens. METHODS: SAMNs were applied for mimicking Fe(3+) ions in solution, acting as magnetically drivable probes to evaluate putative Fe(3+) recognition sites on the microorganism surface. Culture broths and nano-bio-conjugates were characterized by UV-Vis spectroscopy and mass spectrometry. RESULTS: The whole heritage of a membrane porin (OprF) of P. fluorescens Ps_22 cells was recognized and firmly bound by SAMNs. The binding of nanoparticles to OprF porin was correlated to a drastic inhibition of a siderophore (pyoverdine) biosynthesis and to the stimulation of the production and rate of formation of a secondary siderophore. The analysis of metabolic pathways, based on P. fluorescens Ps_22 genomic information, evidenced that this putative secondary siderophore does not belong to a selection of the most common siderophores. CONCLUSIONS: In the scenario of an adhesion mechanism, it is plausible to consider OprF as the biological component deputed to the mineral iron sensing in P. fluorescens Ps_22, as well as one key of siderophore regulation. GENERAL SIGNIFICANCE: The present work sheds light on mineral iron sensing in microorganisms. Peculiar colloidal naked iron oxide nanoparticles offer a useful approach for probing the adhesion of bacterial surface on mineral iron for the identification of the specific recognition site for this iron uptake regulation in microorganisms.
BACKGROUND:Mineraliron(III) recognition by bacteria is considered a matter of debate. The peculiar surface chemistry of novel naked magnetic nanoparticles, called SAMNs (surface active maghemite nanoparticles) characterized by solvent exposed Fe(3+) sites on their surface, was exploited for studying mineraliron sensing in Pseudomonas fluorescens. METHODS:SAMNs were applied for mimicking Fe(3+) ions in solution, acting as magnetically drivable probes to evaluate putative Fe(3+) recognition sites on the microorganism surface. Culture broths and nano-bio-conjugates were characterized by UV-Vis spectroscopy and mass spectrometry. RESULTS: The whole heritage of a membrane porin (OprF) of P. fluorescens Ps_22 cells was recognized and firmly bound by SAMNs. The binding of nanoparticles to OprF porin was correlated to a drastic inhibition of a siderophore (pyoverdine) biosynthesis and to the stimulation of the production and rate of formation of a secondary siderophore. The analysis of metabolic pathways, based on P. fluorescens Ps_22 genomic information, evidenced that this putative secondary siderophore does not belong to a selection of the most common siderophores. CONCLUSIONS: In the scenario of an adhesion mechanism, it is plausible to consider OprF as the biological component deputed to the mineraliron sensing in P. fluorescens Ps_22, as well as one key of siderophore regulation. GENERAL SIGNIFICANCE: The present work sheds light on mineraliron sensing in microorganisms. Peculiar colloidal naked iron oxide nanoparticles offer a useful approach for probing the adhesion of bacterial surface on mineraliron for the identification of the specific recognition site for this iron uptake regulation in microorganisms.
Authors: Luca Fasolato; Nadia Andrea Andreani; Roberta De Nardi; Giulia Nalotto; Lorenzo Serva; Barbara Cardazzo; Stefania Balzan; Lisa Carraro; Federico Fontana; Enrico Novelli Journal: Ital J Food Saf Date: 2018-04-09