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Published on: April 11, 2016
Structural characterization of the immunostimulatory exopolysaccharide produced by Leuconostoc mesenteroides strain
Chiaki Matsuzaki1, Chikahiro Takagaki2, Yusuke Tomabechi1
1Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Suematsu, Nonoichi, Ishikawa 921-8836, Japan.
Abstract:
The exopolysaccharide (EPS) produced by probiotic Leuconostoc mesenteroides subsp. mesenteroides strain NTM048 has been reported to be an immunostimulant that enhances mucosal IgA production. In this study, we found that intranasal administration of mice with the EPS and an antigen (ovalbumin) resulted in secretion of antigen-specific IgA and IgG in the airway mucosa and the serum, suggesting that the EPS has the adjuvant activity for use with mucosal vaccination. Methylation analysis coupled to GC-MS, and 1D and 2D NMR spectroscopy revealed that 94% of the EPS consists of an α-(1 → 6) glucan containing 4% of 1→3-linked α-glucose branches. To determine structures of minor components, we enzymatically digested the glucan with dextranase and used 2D NMR spectroscopy to identify the remaining polymer as a fructan (or fructans), containing both β-(2 → 6)- and β-(2 → 1)-linked fructofuranose residues. These residues may either enter into separate polymers of each linkage type or form a mixed fructan containing both linkage types.
Insights
The exopolysaccharide from Leuconostoc mesenteroides acts as an adjuvant, enhancing immune responses for mucosal vaccines. This probiotic compound boosts IgA and IgG production when administered with antigens.
Area of Science:
- Microbiology
- Immunology
- Carbohydrate Chemistry
Background:
- Probiotic exopolysaccharides (EPS) can stimulate the immune system.
- Leuconostoc mesenteroides subsp. mesenteroides strain NTM048 produces an EPS with known immunostimulant properties.
- Mucosal IgA production is crucial for mucosal immunity.
Purpose of the Study:
- To investigate the adjuvant activity of EPS from NTM048 for mucosal vaccination.
- To elucidate the structural characteristics of the NTM048 EPS.
Main Methods:
- Intranasal administration of EPS and ovalbumin antigen to mice.
- Measurement of antigen-specific IgA and IgG in serum and airway mucosa.
- Methylation analysis coupled with GC-MS.
- 1D and 2D NMR spectroscopy.
- Enzymatic digestion with dextranase.
Main Results:
- Intranasal EPS administration with antigen induced antigen-specific IgA and IgG.
- The EPS is primarily an α-(1→6) glucan (94%) with minor 1→3-linked α-glucose branches (4%).
- Minor components were identified as fructans with β-(2→6) and/or β-(2→1)-linked fructofuranose residues.
Conclusions:
- The EPS from NTM048 exhibits adjuvant activity, enhancing mucosal and systemic immune responses.
- The EPS is a complex carbohydrate composed mainly of glucan with associated fructan structures.
- This EPS holds potential for use in mucosal vaccine formulations.
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