Frece J, Kos B, Svetec I K, Zgaga Z, Mrsa V, Susković J
Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Croatia.
J Appl Microbiol. 2005;98(2):285-92. doi: 10.1111/j.1365-2672.2004.02473.x.
To investigate the functional role of surface layer proteins (S-layer) in probiotic strain Lactobacillus acidophilus M92, especially its influence on adhesiveness to mouse ileal epithelial cells.
Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, ca at 45 kDa in L. acidophilus M92. Southern blot with pBK1 plasmid, containing slpA gene, gave a positive signal, suggesting that L. acidophilus M92 has a slpA gene coding for the S-layer proteins. S-layer proteins of this strain are present during all phases of growth. The S-layer proteins appeared when cells treated with 5 mol l(-1) LiCl were allowed to grow again. Removal of the S-layer proteins reduced adhesion of L. acidophilus M92 to mouse ileal epithelial cells. Furthermore, the viability of cells without S-layer were reduced in simulated gastric juice at low pH range (2, 2.5, 3) and simulated pancreatic juice with bile salts (1.5 and 3 g l(-1)). S-layer proteins of L. acidophilus M92 were resistant to pepsin and pancreatin, in contrast, the treatment with proteinase K led to a significant proteolysis of the S-layer proteins.
These results demonstrated functional role of S-layer; it is responsible for adhesiveness of Lactobacillus acidophilus M92 to mouse ileal epithelial cells and has a protective role for this strain.
S-layer proteins have an important role in the establishment of probiotic strain Lactobacillus acidophilus M92 in the gastrointestinal tract.
研究表层蛋白(S 层)在益生菌嗜酸乳杆菌 M92 中的功能作用,尤其是其对黏附小鼠回肠上皮细胞的影响。
对细胞表面蛋白进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳,结果显示嗜酸乳杆菌 M92 中存在约 45 kDa 的潜在表层(S 层)蛋白。用含有 slpA 基因的 pBK1 质粒进行 Southern 印迹杂交,得到阳性信号,表明嗜酸乳杆菌 M92 具有编码 S 层蛋白的 slpA 基因。该菌株的 S 层蛋白在生长的各个阶段均存在。用 5 mol l(-1) LiCl 处理细胞后再使其生长,S 层蛋白会出现。去除 S 层蛋白会降低嗜酸乳杆菌 M92 对小鼠回肠上皮细胞的黏附。此外,无 S 层的细胞在低 pH 范围(2、2.5、3)的模拟胃液和含胆盐(1.5 和 3 g l(-1))的模拟胰液中的活力会降低。嗜酸乳杆菌 M92 的 S 层蛋白对胃蛋白酶和胰蛋白酶具有抗性,相比之下,用蛋白酶 K 处理会导致 S 层蛋白发生显著的蛋白水解。
这些结果证明了 S 层的功能作用;它负责嗜酸乳杆菌 M92 对小鼠回肠上皮细胞的黏附,并对该菌株具有保护作用。
S 层蛋白在益生菌嗜酸乳杆菌 M92 在胃肠道的定殖中具有重要作用。