Project description:The psychrotolerant Trichococcus patagoniensis was previously reported to produce exopolysaccharides (EPS) that covers the cells, when grown at a low temperature. Although, T. patagoniensis has an optimal growth at 30 ºC, it still can grow at -5 ºC. This capability of T. patagoniensis to grow at subzero temperatures is expected to be reflected in the proteome, membrane lipid composition and/or EPS production. This study aimed to get insight into the cold adaptation mechanisms of T. patagoniensis. We compared the proteomes of cells grown at 0 and 30 ºC and the EPS that is formed.
Project description:Burkholderia cepacia complex bacteria comprise opportunistic pathogens causing chronic respiratory infections in cystic fibrosis (CF) patients. These microorganisms produce exopolysaccharides which are considered virulence determinants. Here, we report the isolation of an evolved nonmucoid variant (17616nmv) after the ancestor, Burkholderia multivorans ATCC 17616, was subjected to prolonged stationary phase. Lack of exopolysaccharide cepacian biosynthesis in the variant was correlated with downregulation of bce genes expression. Furthermore, genome sequence of the variant identified the transposition of a mobile element of the IS406 family into the promoter region of an hns-like gene (Bmul_0158) encoding a histone-like nucleoid structuring protein, a known global transcriptional repressor. This IS element upregulated the expression of Bmul_0158 gene. Transcriptome analysis identified the global effects of this mutation on gene expression, with major changes in motility, pili synthesis, type VI secretion, and chromosome associated functions. Concomitant with these differences, the nonmucoid variant displays reduced adherence to a CF lung bronchial cell line, reduced surface hydrophobicity, forms smaller cellular aggregates, but has an increase in swimming and swarming motilities. Finally, bioinformatic analysis led to the identification of various genomic regions, possibly acquired by horizontal gene transfer, which were transcriptionally repressed by the increased expression of hns gene in the nonmucoid variant. Taken together, our results revealed a significant role for this H-NS protein in the regulation of B. multivorans persistence- and virulence-associated genes.
Project description:The purpose of this course is to study the relationship between the primary structure of exopolysaccharides and the gene expression level of Bifidobacterium longum W13 and W13R, and to better explain the differences between different strains from the level of gene transcription.
Project description:To study their metabolic potential in natural ecosystems, we developed a species-independent LAB microarray, containing 2,269 30-mer oligonucleotides, and targeting 406 genes that play a key role in the production of sugar catabolites, bacteriocins, exopolysaccharides, and aromas, in probiotic and biosafety characteristics, and in stress response. Also, genes linked to negative traits such as antibiotic resistance and virulence are represented. This experiment is a validation experiment, where we hybridized labelled DNA from 20 LAB strains, covering 86% of all oligos. Keywords: Platform validation experiment
Project description:The advances of next-generation sequencing technologies enabled the continuous growth in the number of gene sequences deposited into the CAZy database, meanwhile, the number of biochemically characterized CAZymes is growing at a much slower rate. The biochemical characterization of discovered enzymes is important because of the industrially relevant properties such as high thermal stability or cold-adaptivity that these enzymes may possess. The aim of our project is to screen for up-to-date biochemically uncharacterized CAZymes. We screen for microorganisms that are able to produce extracellular CAZymes. We use LC-MSMS proteomics to identify extracellular enzymes produced by the selected microorganisms.
Project description:we report that succinate, a metabolite abundantly produced by the dysbiotic gut microbiota, induces in vitro biofilm formation of C. difficile strains. We characterized the morphology and spatial composition of succinate- induced biofilms, and compared to non-induced or deoxycholate-induced biofilms, biofilms induced by succinate are significantly thicker, structurally more complex, and poorer in proteins and exopolysaccharides (EPS).
Project description:3-hydroxypropionic acid (3-HP) is a promising platform chemical with various industrial applications. Several metabolic routes to produce 3-HP from organic substrates such as sugars or glycerol have been implemented in yeast, enterobacterial species and other microorganisms. In this work, we investigated 3-HP metabolism of the well-studied ‘Knallgas bacterium’ Cupriavidus necator, a potential C1-chassis for the production of 3-HP and other fatty acid derivatives from CO2 and H2. When testing C. necator for its tolerance towards 3-HP, it was noted that it could utilise the compound as the sole source of carbon and energy.
Project description:To study their metabolic potential in natural ecosystems, we developed a species-independent LAB microarray, containing 2,269 30-mer oligonucleotides, and targeting 406 genes that play a key role in the production of sugar catabolites, bacteriocins, exopolysaccharides, and aromas, in probiotic and biosafety characteristics, and in stress response. Also, genes linked to negative traits such as antibiotic resistance and virulence are represented. This experiment is a validation experiment, where we hybridized labelled DNA from 20 LAB strains, covering 86% of all oligos. Keywords: Platform validation experiment All sampels were hybridized twice in the Cy3 channel. In the Cy5 channel no sample was hybridized.