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Cited by 128Probiotics and Fermented FoodsReproductive tract infections researchBacterial Genetics and Biotechnology

The DOI matches its Crossref record; 75 of the 75 references it cites were found in a registry or an index.

Lactobacillaceae and Cell Adhesion: Genomic and Functional Screening

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Turpin, Williams, Humblot, Christèle, Noordine, Marie-Louise, Thomas, Muriel, Guyot, Jean-Pierre·2012·PLoS ONE
DOI 10.1371/journal.pone.0038034PMC PMC3364998
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Abstract

The analysis of collections of lactic acid bacteria (LAB) from traditional fermented plant foods in tropical countries may enable the detection of LAB with interesting properties. Binding capacity is often the main criterion used to investigate the probiotic characteristics of bacteria. In this study, we focused on a collection of 163 Lactobacillaceace comprising 156 bacteria isolated from traditional amylaceous fermented foods and seven strains taken from a collection and used as controls. The collection had a series of analyses to assess binding potential for the selection of new probiotic candidates. The presence/absence of 14 genes involved in binding to the gastrointestinal tract was assessed. This enabled the detection of all the housekeeping genes (ef-Tu, eno, gap, groEl and srtA) in the entire collection, of some of the other genes (apf, cnb, fpbA, mapA, mub) in 86% to 100% of LAB, and of the other genes (cbsA, gtf, msa, slpA) in 0% to 8% of LAB. Most of the bacteria isolated from traditional fermented foods exhibited a genetic profile favorable for their binding to the gastrointestinal tract. We selected 30 strains with different genetic profiles to test their binding ability to non-mucus (HT29) and mucus secreting (HT29-MTX) cell lines as well as their ability to degrade mucus. Assays on both lines revealed high variability in binding properties among the LAB, depending on the cell model used. Finally, we investigated if their binding ability was linked to tighter cross-talk between bacteria and eukaryotic cells by measuring the expression of bacterial genes and of the eukaryotic MUC2 gene. Results showed that wild LAB from tropical amylaceous fermented food had a much higher binding capacity than the two LAB currently known to be probiotics. However their adhesion was not linked to any particular genetic equipment.

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Crossref · 10.1371/journal.pone.0038034

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Figures

Figure 1 Distribution of genes involved in binding to the gastrointestinal tract in a collection of LAB sampled from starchy fermented foods and in strains used as positive controls. The role of the gene is indicated at the top of the column corresponding to the different strains. The absence of a gene is indicated in white and its presence in black. Sequenced strains are in gray. Strains selected for the adhesion assays and the mucus degrading assays are in black. Genes in L. sakei 23K, L. johnsonii NCC533, and L. acidophilus NCFM were predicted by in silico analysis, except for the cbsA and slpA genes, which were detected on L. acidophilus NCFM by PCR. The dendrogram shows estimated relationships among the strains and was constructed by average-linkage hierarchical analysis using Mev 4.4 software [69].
Figure 1 Distribution of genes involved in binding to the gastrointestinal tract in a collection of LAB sampled from starchy fermented foods and in strains used as positive controls. The role of the gene is indicated at the top of the column corresponding to the different strains. The absence of a gene is indicated in white and its presence in black. Sequenced strains are in gray. Strains selected for the adhesion assays and the mucus degrading assays are in black. Genes in L. sakei 23K, L. johnsonii NCC533, and L. acidophilus NCFM were predicted by in silico analysis, except for the cbsA and slpA genes, which were detected on L. acidophilus NCFM by PCR. The dendrogram shows estimated relationships among the strains and was constructed by average-linkage hierarchical analysis using Mev 4.4 software [69].CC BY · PubMed Central
Figure 2 Ratio of adhered cells to the sum of adhered and non-adhered cells after 2 h incubation at 37°C and the distribution of genes involved in binding to the gastrointestinal tract in the 30 selected LAB. Results are the means ± SD of three independent assays. The ratio of bacteria bound to non-mucus secreting cells (HT29) is in white. The ratio of bacteria bound to mucus secreting cells (HT29-MTX) is in gray. The general role of the gene is indicated at the top left of the line. The absence of a gene is indicated by a “−” and its presence by a “+”. Asterisks indicate sequenced strains of LAB, circles indicate commercial probiotic strains, and squares the strains selected for transcript analysis. Letters indicate a statistical difference in the ratio between the two cell lines (p<0.05, Student-Newman-Keuls test).
Figure 2 Ratio of adhered cells to the sum of adhered and non-adhered cells after 2 h incubation at 37°C and the distribution of genes involved in binding to the gastrointestinal tract in the 30 selected LAB. Results are the means ± SD of three independent assays. The ratio of bacteria bound to non-mucus secreting cells (HT29) is in white. The ratio of bacteria bound to mucus secreting cells (HT29-MTX) is in gray. The general role of the gene is indicated at the top left of the line. The absence of a gene is indicated by a “−” and its presence by a “+”. Asterisks indicate sequenced strains of LAB, circles indicate commercial probiotic strains, and squares the strains selected for transcript analysis. Letters indicate a statistical difference in the ratio between the two cell lines (p<0.05, Student-Newman-Keuls test).CC BY · PubMed Central
Figure 3 Delta A600 24 h after inoculation of reconstituted MRS media containing 2.0% glucose (white) or 0.3% HGM (gray) and residual growth of LAB in MRS with no fermentable carbohydrate. Asterisks indicate sequenced strains of LAB, circles commercial probiotic strains, and squares the strains selected for transcript analysis.
Figure 3 Delta A600 24 h after inoculation of reconstituted MRS media containing 2.0% glucose (white) or 0.3% HGM (gray) and residual growth of LAB in MRS with no fermentable carbohydrate. Asterisks indicate sequenced strains of LAB, circles commercial probiotic strains, and squares the strains selected for transcript analysis.CC BY · PubMed Central
Figure 4 Copy number of mRNA/bacteria of binding related genes in L. paraplantarum 4.4 incubated with HT29 (diagonal hatched bar) or HT29-MTX (vertical hatched bar) and in L. plantarum WCFS1 incubated with HT29 (white) or HT29 MTX (black).
Figure 4 Copy number of mRNA/bacteria of binding related genes in L. paraplantarum 4.4 incubated with HT29 (diagonal hatched bar) or HT29-MTX (vertical hatched bar) and in L. plantarum WCFS1 incubated with HT29 (white) or HT29 MTX (black).CC BY · PubMed Central
Figure 5 Expression of MUC2 in HT29 and HT29-MTX in response to cell binding bacteria. The delta Ct values of MUC2 normalized to the GAPDH gene obtained on HT29 is in white and on the HT29-MTX cell line is in gray. Different letters indicate a statistical difference between the samples (p<0.05, Student-Newman-Keuls test).
Figure 5 Expression of MUC2 in HT29 and HT29-MTX in response to cell binding bacteria. The delta Ct values of MUC2 normalized to the GAPDH gene obtained on HT29 is in white and on the HT29-MTX cell line is in gray. Different letters indicate a statistical difference between the samples (p<0.05, Student-Newman-Keuls test).CC BY · PubMed Central

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Research works printed but not linked: 1

  1. Å Ljungh, 2009, Lactobacillus Molecular Biology: From Genomics to Probiotics Caister Academic Press

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Metadata

Authors
Turpin, W., Humblot, C., Noordine, M.-L., Thomas, M., & Guyot, J.-P.
Affiliations
  • Institut de Recherche pour le Développement (FR)
  • Microbiologie de l’alimentation au service de la santé (FR)
Corresponding author
Humblot, Christèle
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2012-05-31
Volume
7
Issue
5
Pages
e38034
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0038034
PMC
PMC3364998
OpenAlex
W2166637116
Cited by
118
References
75
Citations by year
2026: 3 · 2025: 6 · 2024: 4 · 2023: 7 · 2022: 14 · 2021: 9 · 2020: 6 · 2019: 11 · 2018: 10 · 2017: 6 · 2016: 5 · 2015: 14 · 2014: 15 · 2013: 6
FWCI
17.21
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
  • Life Sciences › Immunology and Microbiology › Microbiology › Reproductive tract infections research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
Lactobacillaceae, lactic acid bacteria, cell adhesion, probiotics, mucus degradation, traditional fermented foods, host-bacteria interactions
MeSH terms
Bacterial Adhesion, Humans, Intestinal Mucosa, Lactobacillaceae, Mucins, Gene Expression Regulation, Bacterial, Genome, Bacterial, HT29 Cells, Mucin-2

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