Characterization of lipoteichoic acid structures from three probiotic Bacillus strains

involvement of d-alanine in their biological activity

Romain Villéger, Naima Saad, Karine Grenier, Xavier Falourd, Loïc Foucat, Maria C. Urdaci, Philippe Bressollier, Tan Sothea Ouk

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Probiotics represent a potential strategy to influence the host’s immune system thereby modulating immune response. Lipoteichoic Acid (LTA) is a major immune-stimulating component of Gram-positive cell envelopes. This amphiphilic polymer, anchored in the cytoplasmic membrane by means of its glycolipid component, typically consists of a poly (glycerol-phosphate) chain with d-alanine and/or glycosyl substitutions. LTA is known to stimulate macrophages in vitro, leading to secretion of inflammatory mediators such as Nitric Oxide (NO). This study investigates the structure–activity relationship of purified LTA from three probiotic Bacillus strains (Bacillus cereus CH, Bacillus subtilis CU1 and Bacillus clausii O/C). LTAs were extracted from bacterial cultures and purified. Chemical modification by means of hydrolysis at pH 8.5 was performed to remove d-alanine. The molecular structure of native and modified LTAs was determined by 1H NMR and GC–MS, and their inflammatory potential investigated by measuring NO production by RAW 264.7 macrophages. Structural analysis revealed several differences between the newly characterized LTAs, mainly relating to their d-alanylation rates and poly (glycerol-phosphate) chain length. We observed induction of NO production by LTAs from B. subtilis and B. clausii, whereas weaker NO production was observed with B. cereus. LTA dealanylation abrogated NO production independently of the glycolipid component, suggesting that immunomodulatory potential depends on d-alanine substitutions. d-alanine may control the spatial configuration of LTAs and their recognition by cell receptors. Knowledge of molecular mechanisms behind the immunomodulatory abilities of probiotics is essential to optimize their use.

Original languageEnglish (US)
Pages (from-to)693-706
Number of pages14
JournalAntonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Volume106
Issue number4
DOIs
StatePublished - Oct 1 2014
Externally publishedYes

Fingerprint

Probiotics
Alanine
Bacillus
Nitric Oxide
Bacillus cereus
Glycolipids
Bacillus subtilis
Macrophages
Molecular Structure
Gas Chromatography-Mass Spectrometry
Immune System
Polymers
Hydrolysis
Cell Membrane
lipoteichoic acid
Bacillus clausii
polyglycerolphosphate

Keywords

  • Immunomodulation
  • Lipoteichoic acid
  • Probiotics
  • Structure–activity relationship

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Characterization of lipoteichoic acid structures from three probiotic Bacillus strains : involvement of d-alanine in their biological activity. / Villéger, Romain; Saad, Naima; Grenier, Karine; Falourd, Xavier; Foucat, Loïc; Urdaci, Maria C.; Bressollier, Philippe; Ouk, Tan Sothea.

In: Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology, Vol. 106, No. 4, 01.10.2014, p. 693-706.

Research output: Contribution to journalArticle

Villéger, Romain ; Saad, Naima ; Grenier, Karine ; Falourd, Xavier ; Foucat, Loïc ; Urdaci, Maria C. ; Bressollier, Philippe ; Ouk, Tan Sothea. / Characterization of lipoteichoic acid structures from three probiotic Bacillus strains : involvement of d-alanine in their biological activity. In: Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology. 2014 ; Vol. 106, No. 4. pp. 693-706.
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