Tannic Acid-Dependent Modulation of Selected Lactobacillus plantarum Traits Linked to Gastrointestinal Survival

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Abstract

Background:Owing to its antimicrobial properties dietary tannins may alter the functional efficacy of probiotic lactobacilli in the gastrointestinal (GI)-tract influencing their growth, viability and molecular adaptation to the intestinal environment.Methods and Findings:The effects of tannic acid on Lactobacillus plantarum WCFS1 were studied by in vitro growth monitoring and visualizing the morphological alteration on the cell wall using transmission electron microscopy. Growth upon tannic acid was characterized by dose-dependent reductions of initial viable counts and extended lag phases. Lag phase-cells growing upon 0.5 mM tannic acid were abnormally shaped and experienced disturbance on the cell wall such as roughness, occasional leakage and release of cell debris, but resumed growth later at tannic acid concentrations high as 2.5 mM. To gain insight on how the response to tannic acid influenced the molecular adaptation of L. plantarum to the GI-tract conditions, gene expression of selected biomarkers for GI-survival was assessed by RT-qPCR on cDNA templates synthetized from mRNA samples obtained from cells treated with 0.5 or 2 mM tannic acid. Tannic acid-dependent gene induction was confirmed for selected genes highly expressed in the gut or with confirmed roles in GI-survival. No differential expression was observed for the pbp2A gene, a biomarker negatively related with GI-survival. However PBP2A was not labeled by Bocillin FL, a fluorescent dye-labeled penicillin V derivative, in the presence of tannic acid which suggests for enhanced GI-survival reportedly associated with the inactivation of this function.Conclusions:Probiotic L. plantarum WCFS1 is able to overcome the toxic effects of tannic acid. This dietary constituent modulates molecular traits linked to the adaptation to intestinal environment in ways previously shown to enhance GI-survival. © 2013 Reverón et al.

Figures

  • Figure 1. Effects of tannic acid on the growth of L. plantarum WCFS1. Cells were grown in RPM media supplemented with increasing tannic acid concentrations. Control, X; 0.1 mM tannic acid, %; 0.25 mM tannic acid, m; 0.5 mM tannic acid, X. CFU, colony-forming units. Representative curves are shown. Values at each time point are the average of at least 2 replica plates. Standard deviations (not shown) were less than 10%. doi:10.1371/journal.pone.0066473.g001
  • Figure 2. Transmission electron micrographs of lag phase-cells of L. plantarum WCFS1 treated with tannic acid. Cells were grown for: 4 h in RPM medium lacking tannic acid (A); 16 h (B, C, D, E, F) in RPM medium containing 0.5 mM tannic acid. Scale bar 0.2 mm (A, B, C, F); 0.5 mm (D, E). doi:10.1371/journal.pone.0066473.g002
  • Table 1. Expression of selected L. plantarum WCFS1 genes upon tannic acid challenge.
  • Figure 3. Effect of tannic acid on PBPs from L. plantarum WCFS1. PBPs were extracted from L. plantarum WCFS1 cells grown in absence (1) or in presence (2) of 0.5 mM tannic acid and labelled with Bocillin FL. These proteins were separated by SDS-PAGE and detected on the gel by fluorography (see Material and methods). Based on their molecular size PBPs were named according to the PBPs annotated in the complete genome sequence of L. plantarum WCFS1 (see text). doi:10.1371/journal.pone.0066473.g003

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Reverón, I., Rodríguez, H., Campos, G., Curiel, J. A., Ascaso, C., Carrascosa, A. V., … de Felipe, F. L. (2013). Tannic Acid-Dependent Modulation of Selected Lactobacillus plantarum Traits Linked to Gastrointestinal Survival. PLoS ONE, 8(6). https://doi.org/10.1371/journal.pone.0066473

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