Project description:To determine whether osmotic pressure affects the translation efficiency of Lactobacillus rhamnosus, the ribosome profiling assay was performed to analyze the changes in translation efficiency in L. rhamnosus ATCC 53103. Under osmotic stress, differentially expressed genes (DEGs) involved in fatty acid biosynthesis and metabolism, ribosome, and purine metabolism pathways were co-regulated with consistent expression direction at translation and transcription levels. DEGs involved in the biosynthesis of phenylalanine, tyrosine, and tryptophan, and the phosphotransferase system pathways also were co-regulated at translation and transcription levels, while they showed opposite expression direction at two levels. Moreover, DEGs involved in the two-component system, amino acid metabolism, and pyruvate metabolism pathways were only regulated at the transcription level. And DEGs involved in fructose and mannose metabolism were only regulated at the translation level. The translation efficiency of DEGs involved in the biosynthesis of amino acids was downregulated while in quorum sensing and PTS pathways was upregulated. In addition, the ribosome footprints accumulated in open reading frame regions resulted in impaired translation initiation and elongation under osmotic stress. In summary, L. rhamnosus ATCC 53103 could respond to osmotic stress by translation regulation and control the balance between survival and growth of cells by transcription and translation.
Project description:Lactobacillus rhamnosus GG has become one of the most widely marketed and studied probiotic strains. Several genes important for probiotic function have been identified, including the spaCBA-srtC1 gene cluster encoding pili, which have been shown to be important for certain of its probiotic properties. The spaCBA-srtC1 gene cluster has been reported to be unstable in L. rhamnosus GG isolated from liquid dairy products and therefore the present study examined the L. rhamnosus GG genome stability throughout an industrial production process from the original deposit to the freeze-dried products including intermediate fermentations and single colony isolates prepared from these samples. The results showed that the original deposit was identical to the reference ATCC and that the genome sequence stayed fully intact throughout the production process. No SNPs or larger genomic changes occurred in any of the samples throughout the production process and the spaCBA-srtC1 gene locus was fully conserved and intact in all 31 samples examined. In addition, phenotypic expression of pili was demonstrated using immune-gold labelling EM. The images showed that pili production was preserved throughout the production process and that the number of pili were consistent in all batches. The present study extends the scope of previous findings to an industrial setting and shows that the region around the spaCBA-srtC1 cluster exhibits high stability in L. rhamnosus GG in an industrial production process.
Project description:Lacticaseibacillus rhamnosus GG (LGG) is a widely consumed probiotic whose potential beneficial effects in humans have been examined in over 250 clinical trials. However, the mechanisms by which LGG modulates host gut physiology remains unknown. R. gnavus is a pathobiont that is strongly associated with inflammatory bowel diseases. Germ free mice were mono-associated with L. rhamonosus (LGG) or R. gnavus (RG) for 21 days, ileum bulk RNA-seq were performed to compare the transcriptomic profiles of LGG or RG associated mice with the transcriptome of germ-free mouse ileum.