Skip to main content
Research article - Peer-reviewed, 2020

Correlation between the Antimicrobial Activity and Metabolic Profiles of Cell Free Supernatants and Membrane Vesicles Produced by Lactobacillus reuteri DSM 17938

Maccelli, Alessandro; Carradori, Simone; Puca, Valentina; Sisto, Francesca; Lanuti, Paola; Crestoni, Maria Elisa; Lasalvia, Alba; Muraro, Raffaella; Bysell, Helena; Di Sotto, Antonella; Roos, Stefan; Grande, Rossella

Abstract

The aim of the work is to assess the antimicrobial activities of Cell Free Supernatants (CFS) and Membrane Vesicles (MVs), produced by Lactobacillus reuteri DSM 17938, versus Gram-positive and Gram-negative bacteria and investigate their metabolic profiles. The Minimum Inhibitory Concentration was determined through the broth microdilution method and cell proliferation assay while the Minimum Bactericidal Concentration was determined by Colony Forming Units counts. The characteristics of the antimicrobial compounds were evaluated by pH adjustments, proteinase treatment, and size fractionation of the CFS. The cytotoxicity of CFS was tested on two human cell lines. A detailed snapshot of the L. reuteri metabolism was attained through an untargeted metabolic profiling by means of high resolution Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS) coupled with Electrospray Ionization Source (ESI). The results showed (i) a greater efficacy of CFS and its fractions towards Gram-negative compared to Gram-positive bacteria; (ii) an antimicrobial effect related to pH-dependent compounds but not to MVs; (iii) a molecular weight < 3 KDa as well as an a non-proteinaceous nature of the antimicrobial compounds; and (iv) more than 200 and 500 putative metabolites annotated in MVs and supernatants, covering several classes of metabolites, including amino acids, lipids, fatty and organic acids, polyalcohols, nucleotides, and vitamins. Some putative compounds were proposed not only as characteristic of specific fractions, but also possibly involved in antimicrobial activity.

Keywords

Lactobacillus reuteri; Limosilactobacillus reuteri; cell free supernatant; metabolomics; extracellular and membrane vesicles; antimicrobial activity; biofilm; probiotics

Published in

Microorganisms
2020, volume: 8, number: 11, article number: 1653

Authors' information

Maccelli, Alessandro
Sapienza University Rome
Carradori, Simone
G d'Annunzio University of Chieti-Pescara
Puca, Valentina
G d'Annunzio University of Chieti-Pescara
Sisto, Francesca
University of Milan
Lanuti, Paola
G d'Annunzio University of Chieti-Pescara
Crestoni, Maria Elisa
Sapienza University Rome
Lasalvia, Alba
Sapienza University Rome
Muraro, Raffaella
G d'Annunzio University of Chieti-Pescara
Bysell, Helena
BioGaia AB
Di Sotto, Antonella
Sapienza University Rome
Swedish University of Agricultural Sciences, Department of Molecular Sciences
Grande, Rossella
G d'Annunzio University of Chieti-Pescara

UKÄ Subject classification

Microbiology

Publication Identifiers

DOI: https://doi.org/10.3390/microorganisms8111653

URI (permanent link to this page)

https://res.slu.se/id/publ/109411