The Biological Role of the S-Layer Produced by Lactobacillus helveticus 34.9 in Cell Protection and Its Probiotic Properties

Iulia-Roxana Angelescu, M. Zamfir, Emanuela-Cătălina Ionetic, S. Grosu-Tudor
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Abstract

Lactobacillus helveticus 34.9 was isolated from a sample of Romanian home-made fermented milk, producing both surface layer proteins and a class III bacteriocin. The present study aimed to investigate the biological and functional role of the S-layer in correlation with its probiotic properties. The presence of S-layer proteins resulted in various degrees of co-aggregation of L. helveticus 34.9 with pathogens and with other lactic acid bacteria, but the removal of these proteins reduced the co-aggregation with all the tested strains. Moreover, the S-layer proved to be involved in cell wall hydrophobicity and cellular protection during freeze-drying. In the simulated passage through the gastrointestinal tract, S-layer depleted cells exhibited increased vulnerability, with greater viability loss in low pH and pepsin treatment compared to control cells. Subsequently, in the small intestine simulation, these cells lost all viability, underscoring the vital role of extracellular proteins for cell protection. The morphological effects of these treatments were observed by scanning electron microscopy. Severe structural damage was noticed when the S-layer was absent, including loss of cell shape and integrity as well as many ghost cells emptied of their content. Finally, the elimination of surface proteins reduced the interaction between L. helveticus 34.9 and mammalian cells.
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螺旋乳杆菌 34.9 产生的 S 层在细胞保护中的生物学作用及其益生菌特性
从罗马尼亚自制的发酵牛奶样本中分离出了螺旋乳杆菌 34.9,它能产生表层蛋白和一种 III 级细菌素。本研究旨在调查 S 层的生物和功能作用与其益生特性的相关性。S 层蛋白的存在导致氦溶乳杆菌 34.9 与病原体和其他乳酸菌发生不同程度的共聚集,但去除这些蛋白后,与所有受试菌株的共聚集程度都有所降低。此外,在冷冻干燥过程中,S 层被证明参与了细胞壁的疏水性和细胞保护。在模拟通过胃肠道的过程中,缺失S层的细胞表现出更大的脆弱性,与对照细胞相比,在低pH值和胃蛋白酶处理中活力损失更大。随后,在小肠模拟中,这些细胞丧失了所有活力,这突出表明了细胞外蛋白质对细胞保护的重要作用。扫描电子显微镜观察了这些处理的形态学影响。当 S 层缺失时,细胞结构受到严重破坏,包括细胞形状和完整性的丧失,以及许多被掏空的幽灵细胞。最后,表面蛋白的消除降低了螺旋藻毒素 34.9 与哺乳动物细胞之间的相互作用。
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