Department of Applied Biology and Chemistry, Faculty of Applied Bio-Science, Tokyo University of Agriculture, Japan.
Division of Agriculture and Agricultural Life Science, The University of Tokyo, Japan.
Microbiology (Reading). 2012 Nov;158(Pt 11):2712-2723. doi: 10.1099/mic.0.060913-0. Epub 2012 Aug 23.
The structural diversity of wall teichoic acid (WTA) was investigated using biochemical and NMR analyses among 19 strains of Lactobacillus plantarum, of which seven were previously established to contain a glycerol-type backbone, whereas the remaining 12 strains possess ribitol-containing WTA. Despite the fact that the WTAs consisted of identical components, namely phosphoric acid, alditol (glycerol or ribitol) and glucose, comparative analysis of the (1)H and (13)C NMR spectra indicated the presence of six different structures, based on the observed differences in the anomeric signals of glucose residues. To determine the six WTA structures, their repeating units were prepared by alkaline hydrolysis, followed by fractionation on HPLC, and analysis by NMR spectroscopy using synthetic molecules as a reference. The structures of the six isolates were established as 1-α-D-glucosyl-sn-glycerol 3-phosphate, 1-α-D-kojibiosyl-sn-glycerol 3-phosphate, 1-α-D-nigerosyl-sn-glycerol 3-phosphate, 4-α-D-kojibiosylribitol 1-phosphate and 1,5-linked di-(2,4-di-α-D-glucosylribitol) phosphate. The backbone structures appeared to be 3,6'-linked poly(1-α-D-glucosyl-sn-glycerol phosphate) for the glycerol-type WTA and 1,5-linked poly(ribitol phosphate) for the ribitol-containing WTA. Moreover, in the analysis of the alkaline hydrolysates on HPLC, only single structures of repeating units were released from each WTA, indicating the high structural uniformity of the WTA in each strain. Notably, analyses of lipoteichoic acid isolated from representative strains harbouring the six different WTAs revealed the universal presence of a 1,3-linked poly(glycerol phosphate) chain, substituted at C-2 of the glycerol residues with glucose residues. These findings provide fundamental information on WTA structural variability in Lb. plantarum, which seems likely to play a pivotal role in the physiology of this bacterial species.
使用生化和 NMR 分析研究了 19 株植物乳杆菌的壁磷壁酸（WTA）的结构多样性，其中 7 株先前被确定为含有甘油型骨架，而其余 12 株含有阿拉伯糖醇型 WTA。尽管 WTA 由相同的成分组成，即磷酸、糖醇（甘油或阿拉伯糖醇）和葡萄糖，但对（1）H 和（13）C NMR 谱的比较分析表明，基于葡萄糖残基的端基信号的差异，存在六种不同的结构。为了确定这六种 WTA 结构，通过碱性水解制备它们的重复单元，然后通过 HPLC 进行分离，并使用合成分子作为参考通过 NMR 光谱进行分析。六种分离株的结构被确定为 1-α-D-葡糖基-sn-甘油 3-磷酸、1-α-D-壳二糖基-sn-甘油 3-磷酸、1-α-D-黑葡糖苷基-sn-甘油 3-磷酸、4-α-D-壳二糖基阿拉伯糖醇 1-磷酸和 1,5-连接的二-（2,4-二-α-D-葡萄糖基阿拉伯糖醇）磷酸。甘油型 WTA 的骨架结构似乎为 3,6'-连接的聚（1-α-D-葡糖基-sn-甘油磷酸），而阿拉伯糖醇型 WTA 的骨架结构为 1,5-连接的聚（阿拉伯糖醇磷酸）。此外，在 HPLC 上对碱性水解产物的分析中，每种 WTA 仅从每个 WTA 释放出单一结构的重复单元，表明每个菌株的 WTA 具有高度的结构均一性。值得注意的是，对代表六种不同 WTA 的菌株分离的脂磷壁酸的分析表明，普遍存在 1,3-连接的聚（甘油磷酸）链，该链在甘油残基的 C-2 上被葡萄糖残基取代。这些发现为植物乳杆菌 WTA 结构变异性提供了基础信息，这似乎在该细菌物种的生理学中起着关键作用。
