Project description:DNA methylation is an important epigenetic modification. DNA methylation phenomenon exists widely in bacteria, plants and animals and is involved in a variety of biological processes.The wild Lactobacillus casei Zhang and its mutant Lactobacillus casei Zhang Δpglx were used as the research subjects.Proteomics was used to explore the effects of DNA methylation on various aspects of Lactobacillus casei.
Project description:Lactobacillus casei Zhang is a probiotic bacterium isolated from koumiss in Inner Mongolia of China that has been commercially used as a starter in the manufacture of dairy products. To study the gene expression profiles of L. casei Zhang during growth in milk, a whole-genome microarray was used. Compared to L. casei Zhang grown to late logarithmic phase in milk, 61 genes were significantly up-regulated (>5 fold) in stationary phase, whereas 26 genes were down-regulated. Collectively, these data showed that the majority of the identified genes was involved in carbohydrate metabolism and energy production, followed by genes involved in nucleotide metabolism, inorganic ion transport, amino acid transport and metabolism, chaperone, etc. This study demonstrates the fundamental effects of cultural conditions on the transcriptome of L. casei Zhang. Moreover, it improves the understanding of the growth and survival mechanism of the bacterium during the late stage of milk fermentation.
Project description:Lactobacillus casei Zhang is a probiotic bacterium isolated from koumiss in Inner Mongolia of China that has been commercially used as a starter in the manufacture of dairy products. To study the gene expression profiles of L. casei Zhang during growth in milk, a whole-genome microarray was used. Compared to L. casei Zhang grown to late logarithmic phase in milk, 61 genes were significantly up-regulated (>5 fold) in stationary phase, whereas 26 genes were down-regulated. Collectively, these data showed that the majority of the identified genes was involved in carbohydrate metabolism and energy production, followed by genes involved in nucleotide metabolism, inorganic ion transport, amino acid transport and metabolism, chaperone, etc. This study demonstrates the fundamental effects of cultural conditions on the transcriptome of L. casei Zhang. Moreover, it improves the understanding of the growth and survival mechanism of the bacterium during the late stage of milk fermentation. L. casei Zhang was grown in milk for 14h (late logarithmic phase) or 18h (stationary phase). 2 biological replicates each.
Project description:Although it is clear that probiotics improve intestinal barrier function, little is known about the effects of probiotics on the aging intestine. We investigated effects of 10-wk bacterial supplementation of Lactobacillus plantarum WCFS1, Lactobacillus casei BL23, or Bifidobacterium breve DSM20213 on gut barrier and immunity in 16-week-old accelerated aging Ercc1-/Δ7 mice, which have a median lifespan of ~20wk, and their wild-type littermates. The colonic barrier in Ercc1-/Δ7 mice was characterized by a thin (<10µm) mucus layer. L. plantarum prevented this decline in mucus integrity in Ercc1-/Δ7 mice, whereas B. breve exacerbated it. Bacterial supplementations affected the expression of immune-related genes, including Toll-like receptor 4. Regulatory T cell frequencies were increased in the mesenteric lymph nodes of L. plantarum- and L. casei-treated Ercc1-/Δ7 mice. L. plantarum- and L. casei-treated Ercc1-/Δ7 mice showed increased specific antibody production in a T cell-dependent immune response in vivo. By contrast, the effects of bacterial supplementation on wild-type control mice were negligible. Thus, supplementation with L. plantarum – but not with L. casei and B. breve – prevented the decline in the mucus barrier in Ercc1-/Δ7 mice. Our data indicate that age is an important factor influencing beneficial or detrimental effects of candidate probiotics. These findings also highlight the need for caution in translating beneficial effects of probiotics observed in young animals or humans to the elderly.
Project description:Lactobacillus casei Zhang is a probiotic bacterium isolated from koumiss in Inner Mongolia of China. Gene expression dynamics of L. casei Zhang during growth in soymilk was investigated in attempt to reveal the mechanisms involved in growth stimulation for growing probiotics in. Comparison of different transcripts next to each other revealed 162 and 63 significantly induced genes in late logarithmic phase and stationary phase, of which the expression was at least 3 fold up-regulated and down-regulated, respectively. Approximately, 38.4% of the up-regulated genes were associated with amino acid transport and metabolism notably for histidine and lysine biosynthesis, followed by genes/gene clusters involved in carbohydrate transport and metabolism, lipid transport and metabolism, and inorganic ion transport and metabolism. The analysis results suggest that the stimulatory effect of soymilk-based ecosystem on the L. casei Zhang growth is more complex than amino acids or oligopeptides supply.
Project description:Lactobacillus casei Zhang is a probiotic bacterium isolated from koumiss in Inner Mongolia of China. Gene expression dynamics of L. casei Zhang during growth in soymilk was investigated in attempt to reveal the mechanisms involved in growth stimulation for growing probiotics in. Comparison of different transcripts next to each other revealed 162 and 63 significantly induced genes in late logarithmic phase and stationary phase, of which the expression was at least 3 fold up-regulated and down-regulated, respectively. Approximately, 38.4% of the up-regulated genes were associated with amino acid transport and metabolism notably for histidine and lysine biosynthesis, followed by genes/gene clusters involved in carbohydrate transport and metabolism, lipid transport and metabolism, and inorganic ion transport and metabolism. The analysis results suggest that the stimulatory effect of soymilk-based ecosystem on the L. casei Zhang growth is more complex than amino acids or oligopeptides supply. To study gene expression dynamics of L. casei Zhang during growth in soymilk, a whole genome microarray was used to screen for differentially expressed genes when grown to lag phase, late logarithmic phase, and stationary phase.
Project description:Nutrient starvation is an important survival challenge for bacteria during industrial production of functional foods. Lactobacilli are increasingly being used as probiotics in functional foods. As next-generation sequencing technology has greatly advanced, we performed integrative proteomic and genomic analysis to investigate the response of Lactobacillus casei Zhang to a glucose-restricted environment. L. casei Zhang strains were permitted to evolve in glucose-limited or normal medium from a common ancestor over a 3-year period, and they were sampled after 1000, 2000, 3000, 4000, 5000, 6000, 7000, and 8000 generations and subjected to proteomic and genomic analyses. Genomic resequencing data revealed different point mutations and other mutational events in each generation of L. casei Zhang under glucose limitation stress. The proteins expressed differentially under glucose limitation were found to be significantly related to fructose and mannose metabolism, carbohydrate metabolic processes, lyase activity, and amino acid-transporting ATPase activity. The integrative proteomic and genomic analysis revealed that the mutations protected L. casei Zhang against glucose starvation by regulating other cellular carbohydrate, fatty acid, and amino acid catabolism; phosphoenolpyruvate system pathway activation; glycogen synthesis; ATP consumption; pyruvate metabolism; and general stress response protein expression. The results help reveal the mechanisms of adapting to glucose starvation and provide new strategies for enhancing the industrial utility of L. casei Zhang.
Project description:The survival of probiotics could be strongly enhanced by delivery vehicles, but the mechanism was unknown. In this study, whole genome microarray technology was used to detect the gene expression profiles of probiotic Lactobacillus casei Zhang in the absence and presence of fermented milk in simulated gastrointestinal tract. The results showed the gene expression profiles were significantly different under these two different conditions. The change of the gene expression profile may be helpful to comprehend the role of delivery vehicles in enhancing the survival of probiotics. Twelve samples of bacterial cells in the absence and presence of fermented milk were collected at 3 h in simulated gastric juice, 4 h and 8 h in intestinal juice, 2 biological replicates were obtained in each time point. The average of the normalized expression values of the 2 biological replicates for each probe was regarded as the expression value of a predicted gene. To identify the expression pattern of each gene across different treatments, n-fold change ratios were calculated for every gene in pure L. casei Zhang versus L. casei Zhang in fermented milk.