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Extracellular Vesicles Produced by Bifidobacterium longum Export Mucin-Binding Proteins

机译：通过双歧杆菌产出粘膜结合蛋白产生的细胞外囊泡

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Extracellular proteins are important factors in host-microbe interactions; however, the specific factors that enable bifidobacterial adhesion and survival in the gastrointestinal (GI) tract are not fully characterized. Here, we discovered that Bifidobacterium longum NCC2705 cultured in bacterium-free supernatants of human fecal fermentation broth released a myriad of particles into the extracellular environment. The aim of this study was to characterize the physiological properties of these extracellular particles. The particles, approximately 50 to 80?nm in diameter, had high protein and double-stranded DNA contents, suggesting that they were extracellular vesicles (EVs). A proteomic analysis showed that the EVs primarily consisted of cytoplasmic proteins with crucial functions in essential cellular processes. We identified several mucin-binding proteins by performing a biomolecular interaction analysis of phosphoketolase, GroEL, elongation factor Tu (EF-Tu), phosphoglycerate kinase, transaldolase (Tal), and heat shock protein 20 (Hsp20). The recombinant GroEL and Tal proteins showed high binding affinities to mucin. Furthermore, the immobilization of these proteins on microbeads affected the permanence of the microbeads in the murine GI tract. These results suggest that bifidobacterial exposure conditions that mimic the intestine stimulate B. longum EV production. The resulting EVs exported several cytoplasmic proteins that may have promoted B. longum adhesion. This study improved our understanding of the Bifidobacterium colonization strategy in the intestinal microbiome.IMPORTANCE Bifidobacterium is a natural inhabitant of the human gastrointestinal (GI) tract. Morphological observations revealed that extracellular appendages of bifidobacteria in complex microbial communities are important for understanding its adaptations to the GI tract environment. We identified dynamic extracellular vesicle (EV) production by Bifidobacterium longum in bacterium-free fecal fermentation broth that was strongly suggestive of differing bifidobacterial extracellular appendages in the GI tract. In addition, export of the adhesive moonlighting proteins mediated by EVs may promote bifidobacterial colonization. This study provides new insight into the roles of EVs in bifidobacterial colonization processes as these bacteria adapt to the GI environment.
机译：细胞外蛋白是宿主微生物相互作用的重要因素;然而，在胃肠道（GI）道中能够使双歧杆菌粘附和存活的特定因素没有完全表征。在这里，我们发现在人粪便发酵液的无细菌上清液中培养的双歧杆菌NCC2705释放了颗粒的无数颗粒进入细胞外环境。本研究的目的是表征这些细胞外颗粒的生理性质。直径大约50至80μm的颗粒具有高蛋白质和双链DNA含量，表明它们是细胞外囊（EVS）。蛋白质组学分析表明，EVS主要由细胞质蛋白质组成，具有主要细胞过程中具有至关重要的功能。我们通过进行磷酸溶胶酶，腹股沟，伸长因子Tu（EF-Tu），磷酸甘油糖激酶，甲蛋白（TAL）和热休克蛋白20（HSP20）的生物分子相互作用分析来鉴定几种粘蛋白结合蛋白。重组腹股沟和术蛋白对粘蛋白显示出高结合亲和力。此外，对微珠上的这些蛋白质的固定影响影响了小鼠Gi道中微珠的持续时间。这些结果表明，模拟肠刺激B. Longum EV生产的双歧杆菌暴露条件。得到的EVS导出了几种可促进B. Longum粘附的细胞质蛋白质。本研究改善了我们对肠道微生物组成的双歧杆菌殖民化策略的理解。分别双歧杆菌是人胃肠道（GI）道的自然居民。形态学观察表明，复杂的微生物群体中双歧杆菌的细胞外阑尾都很重要，对理解其对GI道环境的适应性。我们鉴定了在无菌粪便发酵肉芽中的双歧杆菌延长的动态细胞外囊泡（EV）产生，这强烈暗示了GI沟槽中的不同双歧杆菌细胞外阑尾。此外，EVS介导的粘合态度的蛋白质的出口可能促进双歧杆菌定植。本研究为这些细菌适应GI环境，对EVS在双歧杆菌定植过程中的作用提供了新的洞察力。

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