Matsuzaki Chiaki, Takagaki Chikahiro, Tomabechi Yusuke, Forsberg Lennart S, Heiss Christian, Azadi Parastoo, Matsumoto Kenji, Katoh Toshihiko, Hosomi Koji, Kunisawa Jun, Yamamoto Kenji, Hisa Keiko
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Suematsu, Nonoichi, Ishikawa 921-8836, Japan.
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Suematsu, Nonoichi, Ishikawa 921-8836, Japan; Laboratory of Vaccine Materials, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
Carbohydr Res. 2017 Aug 7;448:95-102. doi: 10.1016/j.carres.2017.06.004. Epub 2017 Jun 15.
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.
据报道，益生菌嗜温明串珠菌肠膜明串珠菌亚种NTM048产生的胞外多糖（EPS）是一种免疫刺激剂，可增强粘膜IgA的产生。在本研究中，我们发现给小鼠鼻内注射EPS和抗原（卵清蛋白）会导致气道粘膜和血清中分泌抗原特异性IgA和IgG，这表明EPS具有用于粘膜疫苗接种的佐剂活性。结合GC-MS的甲基化分析以及一维和二维NMR光谱显示，94%的EPS由含有4% 1→3连接的α-葡萄糖分支的α-(1→6)葡聚糖组成。为了确定次要成分的结构，我们用葡聚糖酶对葡聚糖进行酶解，并使用二维NMR光谱将剩余的聚合物鉴定为果聚糖，其含有β-(2→6)-和β-(2→1)-连接的呋喃果糖残基。这些残基可能进入每种连接类型的单独聚合物中，或者形成包含两种连接类型的混合果聚糖。
