Mucin- and carbohydrate-stimulated adhesion and subproteome changes of the probiotic bacterium Lactobacillus acidophilus NCFM
Mucin- and carbohydrate-stimulated adhesion and subproteome changes of the probiotic bacterium Lactobacillus acidophilus NCFM
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DOI：
10.1016/j.jprot.2017.05.015
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发表时间：
2017-06-23
影响因子：
3.3
通讯作者：
Svensson, Birte
中科院分区：
文献类型：
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作者：
Celebioglu, Hasan Ufuk;Olesen, Sita Vaag;Svensson, Birte
对肠粘膜的粘附是益生菌的重要特性。粘附被认为可以增加宿主细菌的相互作用，从而可能给宿主带来健康益处。研究了益生菌嗜酸乳杆菌 NCFM 与粘附和表面蛋白质组变化相关的分子事件，该益生菌以已建立或新兴的益生元碳水化合物作为碳源，并在粘蛋白（上皮粘液层的糖蛋白）存在下培养。对 HT29 细胞和粘蛋白的粘附变化与碳源和粘蛋白诱导的亚蛋白质组丰度差异相关。具体来说，虽然低聚果糖 (FOS) 上的生长仅刺激对肠道 HT-29 细胞的粘附，但纤维二糖和聚葡萄糖还增加了对粘蛋白的粘附。对于在补充粘蛋白的葡萄糖上生长的细菌来说，对 HT-29 细胞的粘附增加了约 2 倍。比较 2DE-MS 表面蛋白质组分析显示，表面存在不同的能量代谢蛋白质，表明它们发挥兼职功能。添加粘蛋白的细菌的丙酮酸激酶和果糖二磷酸醛缩酶的相对丰度增加了约2倍，而相对丰度降低了32-2.1倍的6个点包括延伸因子G、磷酸甘油酸激酶、BipAEFTU家族GTP结合蛋白、核糖核苷三磷酸还原酶、腺苷酸琥珀酸合成酶、30S核糖体蛋白S1和锰依赖性无机焦磷酸酶。与葡萄糖生长的嗜酸乳杆菌 NCFM 相比，纤维二糖表面蛋白质组的磷酸盐饥饿诱导蛋白应激相关蛋白、热稳定支链淀粉酶和延伸因子 G 增加了 4.4-2.4 倍，而 GAPDH、延伸因子 Ts 和丙酮酸激酶的相对丰度减少了 2.0-1.5 倍。将重组嗜酸乳杆菌 NCFM 延伸因子 G 和丙酮酸激酶添加到包被的粘蛋白层中，可显着抑制细菌随后的粘附。 生物学意义：人类饮食对于肠道健康和食物成分很重要，尤其是不可消化的碳水化合物可以有益地改变微生物群。在本研究中，通过测试对粘蛋白层和肠道细胞的粘附力，并将其与被认为对宿主相互作用很重要的表面蛋白丰度的变化进行比较，研究了新兴和已建立的益生元碳水化合物对嗜酸乳杆菌 NCFM 益生菌潜力的影响。用低聚果糖、纤维二糖或聚葡萄糖以及补充粘蛋白的葡萄糖作为碳源培养细菌后，观察到粘附力增加。增强的粘附能力可以延长细菌在胃肠道中的停留时间，从而产生积极的健康影响。某些表面蛋白在各种条件下（即在纤维二糖或补充粘蛋白的葡萄糖上生长）的相对丰度较高，表明这些蛋白参与粘附，这通过两种重组产生的兼职蛋白的竞争所证实。嗜酸乳杆菌 NCFM 与不同碳水化合物的组合揭示了合生元相互作用的潜在细菌决定因素，包括刺激粘附。 2017 Elsevier B.V. 保留所有权利。
Adhesion to intestinal mucosa is a crucial property for probiotic bacteria. Adhesion is thought to increase host bacterial interactions, thus potentially enabling health benefits to the host. Molecular events connected with adhesion and surface proteome changes were investigated for the probiotic Lactobacillus acidophilus NCFM cultured with established or emerging prebiotic carbohydrates as carbon source and in the presence of mucin, the glycoprotein of the epithelial mucus layer. Variation in adhesion to HT29-cells and mucin was associated with carbon source and mucin-induced subproteome abundancy differences. Specifically, while growth on fructooligosaccharides (FOS) only stimulated adhesion to intestinal HT-29 cells, cellobiose and polydextrose in addition increased adhesion to mucin. Adhesion to HT-29 cells increased by about 2-fold for bacteria grown on mucin-supplemented glucose. Comparative 2DE-MS surface proteome analysis showed different proteins in energy metabolism appearing on the surface, suggesting they exert moonlighting functions. Mucin-supplemented bacteria had relative abundance of pyruvate kinase and fructose-bisphosphate aldolase increased by about 2-fold while six spots with 32-2.1 fold reduced relative abundance comprised elongation factor G, phosphoglycerate kinase, BipAEFTU family GTP-binding protein, ribonucleoside triphosphate reductase, adenylosuccinate synthetase, 30S ribosomal protein S1, and manganese-dependent inorganic pyrophosphatase. Surface proteome of cellobiose-compared to glucose-grown L. acidophilus NCFM had phosphate starvation inducible protein stress-related, thermostable pullulanase, and elongation factor G increasing 4.4-2.4 fold, while GAPDH, elongation factor Ts, and pyruvate kinase were reduced by 2.0-1.5 fold in relative abundance. Addition of recombinant L. acidophilus NCFM elongation factor G and pyruvate kinase to a coated mucin layer significantly suppressed subsequent adhesion of the bacterium.Biological significance: Human diet is important for intestinal health and food components, especially non-digestible carbohydrates can beneficially modify the microbiota. In the present study, effects of emerging and established prebiotic carbohydrates on the probiotic potential of Lactobacillus acidophilus NCFM were investigated by testing adhesion to a mucin layer and intestinal cells, and comparing this with changes in abundancy of surface proteins thought to be important for host interactions. Increased adhesion was observed following culturing of the bacterium with fructooligosaccharides, cellobiose or polydextrose, as well as mucin-supplemented glucose as carbon source. Enhanced adhesion ability can prolong bacterial residence in GIT yielding positive health effects. Higher relative abundance of certain surface proteins under various conditions (i.e. grown on cellobiose or mucin-supplemented glucose) suggested involvement of these proteins in adhesion, as confirmed by competition in case of two recombinantly produced moonlighting proteins. Combination of Lactobacillus acidophilus NCFM with different carbohydrates revealed potential bacterial determinants of synbiotic interactions, including stimulation of adhesion. 2017 Elsevier B.V. All rights reserved.