Project description:The bacterial cell envelope is a complex, multilayered structure that is not only essential to maintain cellular integrity, but also facilitates vital bacterial processes such as adaption, colonization and adhesion. Cell envelopes comprise a wide range of molecules, such as proteins and capsular polysaccharide (CPS), which collectively decorate the bacterial cell in a species- and strain specific manner. Here we characterized the 4 CPS gene cluster of L. plantarum and assessed the impact on CPS (combinatorial) gene deletions on surface polysaccharide composition. individual KOs of the cps clusters and the combination of all four together were compared with the WT expression in a loop design. The KO of 1-3 was directly compared to the WT (dye swapped)
Project description:The bacterial cell envelope is a complex, multilayered structure that is not only essential to maintain cellular integrity, but also facilitates vital bacterial processes such as adaption, colonization and adhesion. Cell envelopes comprise a wide range of molecules, such as proteins and capsular polysaccharide (CPS), which collectively decorate the bacterial cell in a species- and strain specific manner. Here we characterized the 4 CPS gene cluster of L. plantarum and assessed the impact on CPS (combinatorial) gene deletions on surface polysaccharide composition.
Project description:Whole genome transcriptional profiling was used to characterize the response of Lactobacillus plantarum WCFS1 human isolate during challenge with oleuropein. Twelve independent experiments were performed and mixed at random in groups of four for total of three RNA samples. The transcriptional profile shows that Lactobacillus plantarum WCFS1 adapts its metabolic capacity to acquire certain carbohydrates and repress the expression of genes involved in fatty acid biosyntheis. The transcriptomic datasets also revealed the downregulation of genes related to the biosynthesis of capsular polysaccharides and genes coding for ABC-type transporters. In addition, induction of oligopeptide permeases is also part of the response of Lactobacillus plantarum WCFS1 to oleuropein.
Project description:Identification of proteins contained in extracellular vesicles of Lactiplantibacillus plantarum PCM 2675. Dataset is related to publication http://dx.doi.org/10.20517/evcna.2024.49. This work was financially supported by the National Science Centre, Poland (no. 2021/43/D/NZ6/01464).
Project description:Probiotics have been suggested to ameliorate the function of the intestinal epithelial barrier and so have several mediators and receptors of the expanded endocannabinoid system, the endocannabinoidome (eCBome). Here we cocultured three live strains of Lactiplantibacillus plantarum with intestinal epithelial organoids to study their effects on the gut barrier function and the possible involvement of the eCBome in this effect. All three L.plantarum strains variously reduced the trans-epithelial permeability of intestinal organoids and promoted increased mRNA expression of several tight junction proteins and intestinal barrier proteins. Concomitantly, the three strains upregulated the expression of genes encoding biosynthetic enzymes (i.e., NapePLD, Abdh4, Gde1, Daglb) and receptors (i.e., Cnr1, Cnr2, Gpr55, and Ppara), while concurrently downregulating the expression of two essential catabolic enzymes (i.e. Faah and Naaa), involved in the signaling of several eCBome mediators known for their role in regulating the intestinal epithelial barrier. Selective inhibitors of eCBome mediator degrading enzymes FAAH and MAGL, i.e., URB597 and JZL184, increased N-acyl-ethanolamine (NAE) and 2-monoacylglycerol (2-MAG) levels, respectively, enhanced the expression of intestinal epithelial barrier genes and reduced the trans-epithelial permeability of organoids, as for L. plantarum strains. Interestingly, inflammation-induced trans-epithelial permeability in organoids was also reversed by both FAAH and MAGL inhibitors. We surmise that elevated endogenous levels of either NAEs or 2-MAGs promote improvement in small intestine trans-epithelial permeability and that L. plantarum strains may exploit this mechanism to promote these beneficial effects.
Project description:To characterize the effect of lactic acid on the L. plantarum growth and adaptation, we investigated the transcriptome under hydrochloride (HCl) or lactic acid at the early stage of the growth.
Project description:Cyclic di-adenosine monophosphate (c-di-AMP) is a crucial second messenger used by bacteria to mediate bacterial-host interactions. However, its roles in Lactiplantibacillus plantarum remain poorly understood. Here, we identify RwpL, which consists solely of an HTH domain, as a c-di-AMP receptor in L. plantarum WCFS1. RwpL mediates epithelial adhesion of L. plantarum WCFS1 and functions as both a transcription inhibitor and activator, with transcriptional output determined by the position of its binding site relative to the transcription start site (TSS) of target genes. Specifically, RwpL inhibits wxlA transcription by binding downstream of its TSS, thus reducing WxlA interactions with fibronectin, a matrix protein on the surface of intestinal epithelia. In contrast, RwpL activates transcription of the matrix-degrading metalloprotease gene mmpL by recruiting the sigma factor RpoA upstream of its TSS. Notably, elevated c-di-AMP levels diminish RwpL binding affinity for both DNA and RpoA, thereby reversing its regulatory functions and enhancing L. plantarum adhesion to host epithelial cells. Together, our findings uncover a dual and reversible regulatory mechanism through which c-di-AMP modulates bacterial-host interactions in L. plantarum.