do Carmo Fillipe L R, Silva Wanderson M, Tavares Guilherme C, Ibraim Izabela C, Cordeiro Barbara F, Oliveira Emiliano R, Rabah Houem, Cauty Chantal, da Silva Sara H, Canário Viana Marcus V, Caetano Ana C B, Dos Santos Roselane G, de Oliveira Carvalho Rodrigo D, Jardin Julien, Pereira Felipe L, Folador Edson L, Le Loir Yves, Figueiredo Henrique C P, Jan Gwénaël, Azevedo Vasco
Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Institut National de la Recherche Agronomique, UMR1253 Science & Technologie du Lait & de l'Oeuf, Rennes, France.
Front Microbiol. 2018 Aug 17;9:1807. doi: 10.3389/fmicb.2018.01807. eCollection 2018.
is a beneficial Gram-positive bacterium, traditionally used as a cheese-ripening starter, and currently considered as an emerging probiotic. As an example, the CIRM-BIA 129 strain recently revealed promising immunomodulatory properties. Its consumption accordingly exerts healing effects in different animal models of colitis, suggesting a potent role in the context of inflammatory bowel diseases. This anti-inflammatory effect depends on surface layer proteins (SLPs). SLPs may be involved in key functions in probiotics, such as persistence within the gut, adhesion to host cells and mucus, or immunomodulation. Several SLPs coexist in CIRM-BIA 129 and mediate immunomodulation and adhesion. A mutant CIRM-BIA 129Δ (CB129Δ) strain was shown to exhibit decreased adhesion to intestinal epithelial cells. In the present study, we thoroughly analyzed the impact of this mutation on cellular properties. Firstly, we investigated alterations of surface properties in CB129Δ. Surface extractable proteins, surface charges (ζ-potential) and surface hydrophobicity were affected by the mutation. Whole-cell proteomics, using high definition mass spectrometry, identified 1,288 quantifiable proteins in the wild-type strain, i.e., 53% of the theoretical proteome predicted according to CIRM-BIA 129 genome sequence. In the mutant strain, we detected 1,252 proteins, including 1,227 proteins in common with the wild-type strain. Comparative quantitative analysis revealed 97 proteins with significant differences between wild-type and mutant strains. These proteins are involved in various cellular process like signaling, metabolism, and DNA repair and replication. Finally, analysis predicted that gene is not part of an operon, thus not affecting the downstream genes after gene knockout. This study, in accordance with the various roles attributed in the literature to SLPs, revealed a pleiotropic effect of a single mutation, in the probiotic . This suggests that SlpB may be at a central node of cellular processes and confirms that both nature and amount of SLPs, which are highly variable within the species, determine the probiotic abilities of strains.
是一种有益的革兰氏阳性细菌，传统上用作奶酪成熟发酵剂，目前被视为一种新兴的益生菌。例如，CIRM-BIA 129菌株最近显示出有前景的免疫调节特性。因此，食用它在不同的结肠炎动物模型中发挥治愈作用，表明在炎症性肠病的背景下具有重要作用。这种抗炎作用取决于表层蛋白（SLPs）。SLPs可能参与益生菌的关键功能，如在肠道内的持久性、与宿主细胞和黏液的黏附或免疫调节。几种SLPs共存于CIRM-BIA 129中，并介导免疫调节和黏附。一个突变的CIRM-BIA 129Δ（CB129Δ）菌株显示出对肠上皮细胞的黏附减少。在本研究中，我们全面分析了这种突变对细胞特性的影响。首先，我们研究了CB129Δ表面特性的改变。表面可提取蛋白、表面电荷（ζ电位）和表面疏水性受到突变的影响。使用高分辨率质谱的全细胞蛋白质组学在野生型菌株中鉴定出1288种可定量的蛋白质，即根据CIRM-BIA 129基因组序列预测的理论蛋白质组的53%。在突变菌株中，我们检测到1252种蛋白质，其中包括与野生型菌株共有的1227种蛋白质。比较定量分析揭示了野生型和突变菌株之间有97种蛋白质存在显著差异。这些蛋白质参与各种细胞过程，如信号传导、代谢以及DNA修复和复制。最后，分析预测基因不是操纵子的一部分，因此在基因敲除后不会影响下游基因。本研究根据文献中赋予SLPs的各种作用，揭示了益生菌中单个突变的多效性作用。这表明SlpB可能处于细胞过程的中心节点，并证实了在该物种中高度可变的SLPs的性质和数量决定了菌株的益生菌能力。
