Project description:We present a study combining gene expression analyses and bioinformatic assessment. 1120 sequences annotated as sulfatase encoding from eight Rhodopirellula strains were clustered into 173 groups of orthologous and paralogous genes. A selection of 709 sulfatase gene strains were aligned to 66 sulfatase genes of known function to check for their respective substrate specificity. Thereby, 22 major phylogenetic clusters were observed with just five being mixed clusters between Rhodopirellula and reference sequences. This indicates a huge diversity on the substrate recognition level, unexpected from conventional annotations in public databases. Exemplarily, Rhodopirellula baltica SH1T was grown on different sulfated polysaccharides. Subsequent gene expression analyses using whole genome microarrays revealed distinct sulfatase expression profiles based on substrates tested. Expression profiles strongly point towards a functional link between sulfated polysaccharides and sulfatases. Moreover, further potential functions can be deduced from the expression profiles. In silico assessment backed up in vivo findings and raised the question, whether related strains and species are even more adapted to utilizing sulfated polysaccharides present in marine environments. Dual channel microarrays have been used for comparing the gene expression of R. baltica SH1T under reference condition (grown on glucose) and grown on sulfated polysaccharides as substrates of interest. Substrates tested have been: Chondroitin sulfate, lambda carrageenan and fucoidan. Control: 3 Biological replicates, Substrates tested: 2-3 Biological replicates, 2-3 replicates per array
Project description:We present a study combining gene expression analyses and bioinformatic assessment. 1120 sequences annotated as sulfatase encoding from eight Rhodopirellula strains were clustered into 173 groups of orthologous and paralogous genes. A selection of 709 sulfatase gene strains were aligned to 66 sulfatase genes of known function to check for their respective substrate specificity. Thereby, 22 major phylogenetic clusters were observed with just five being mixed clusters between Rhodopirellula and reference sequences. This indicates a huge diversity on the substrate recognition level, unexpected from conventional annotations in public databases. Exemplarily, Rhodopirellula baltica SH1T was grown on different sulfated polysaccharides. Subsequent gene expression analyses using whole genome microarrays revealed distinct sulfatase expression profiles based on substrates tested. Expression profiles strongly point towards a functional link between sulfated polysaccharides and sulfatases. Moreover, further potential functions can be deduced from the expression profiles. In silico assessment backed up in vivo findings and raised the question, whether related strains and species are even more adapted to utilizing sulfated polysaccharides present in marine environments.
Project description:Brown algae synthesize various polysaccharides that are ultimately catabolized by marine heterotrophic bacteria. Complex cell wall polysaccharides such as sulfated fucans are considered recalcitrant to microbial degradation and their pathways remain elusive. The branched structure of fucans sterically constraints enzyme-substrate interaction and also, fucan structure varies depending on algae and season challenging adaptation of microbial pathways. Here we show how Lentimonas specialized to overcome the complexity and diversity of sulfated fucans. The strain acquired a 0.9 mbp plasmid with over 200 glycoside hydrolases and sulfatases for the degradation of at least six different sulfated fucans. Per fucan, 100 enzymes are induced and we identified three structural types of fucans with similar pathways depending on their galactose, acetate and sulfate content. The highly decorated structure sulfated fucans expands the copy number and diversity of few key enzyme families, namely GH29, GH95, GH141 and sulfatases S1_15, S1_16 and S1_17. Those enzymes are co-regulated in large operons to step-wise degrade sulfated fucans. Fucose metabolism places additional burden as the conversion of toxic intermediates into lactate and propanediol occurs in a proteome-costly bacterial microcompartment. Through analyzing available genomes and metagenomes, we emphasize that Verrucomicrobia are abundant, yet specialized degraders of complex polysaccharides.
Project description:This pilot research trial studies the effects of chemotherapy on intestinal bacteria/organisms (microbiota) in patients newly diagnosed with breast cancer. Change in intestinal microbiota may be associated with weight gain in patients treated with chemotherapy. Weight gain has been also associated with cancer recurrence. Examining the types and quantity of bacterial composition in the stool of breast cancer patients treated with chemotherapy may help determine whether body weight and composition are associated with changes in the intestinal microbiota and allow doctors to plan better treatment to prevent weight gain and possibly disease recurrence.
Project description:Gut microbiota participates in diverse metabolic and homeostatic functions related to health and well-being. Individual variation in its composition depends on many factors including dietary factors. We profiled enzymatic activity of fecal microbiota in 63 healthy adult individuals using metaproteomics, and identified Bacteroides and Prevotella –derived microbial CAZy (carbohydrate-active) enzymes involved in glycan foraging. One particular profile with many Bacteroides-derived CAZy was identified in one-third of subjects (n=20), and it associated with high abundancy of Bacteroides in most subjects. In other subjects (n=8) with dietary parameters similar to former, microbiota showed intense expression of Prevotella-derived CAZy including exo−beta−(1,4)−xylanase, xylan-1,4−beta−xylosidase, alpha−L−arabinofuranosidase and several other CAZy belonging to glycosyl hydrolase families involved in digestion of complex plant-derived polysaccharides. This associated invariably with robust representation of Prevotella in gut microbiota, while subjects with intermediate representation of Prevotella showed no CAZy profile. Identification of Bacteroides- and Prevotella-derived CAZy in microbiota proteome and their association with robust differences in microbiota composition, the latter with exceptionally high Prevotella abundancy in the gut, are in evidence of individual variation in metabolic adaptation of gut microbiota with an impact on colonizing competence.
Project description:Heparan sulfate proteoglycans (HSPGs) are expressed on virtually all animal cells and are involved in many important biological processes. Each HSPG consists of a core protein with one or more covalently attached linear heparan sulfate (HS) chains composed of alternating glucosamine and uronic acids that are heterogeneously N- and O-sulfated. The arrangement and orientation of the sulfated sugar residues of HS specify the location of distinct ligand binding sites on the cell surface, and these modifications can vary temporally during development and spatially across tissues. While most of the enzymes involved in HS biosynthesis have been studied extensively, much less information exists regarding the specific mechanisms that give rise to the variable composition and binding properties of HS.
Project description:Phytosulfokine (PSK) is a plant peptide hormone that contributes to plant signaling and induced various of effects, including growth, senescence, stress tolerant and defense responses. PSK is a pentapeptide with two tyrosine sulfated by tyrosylprotein sulfotransferase (TPST) enzyme. The sulfated PSK can bind to its cell surface receptors, but the transcriptional readouts remain largely unknown. Here, we treated tpst mutant that are unable to produce native sulfated PSK with synthetic active PSK to capture concentration-sensitive readouts of PSK signaling.
Project description:Phytosulfokine (PSK) is a plant peptide hormone that contributes to plant signaling and induced various of effects, including growth, senescence, stress tolerant and defense responses. PSK is a pentapeptide with two tyrosine sulfated by tyrosylprotein sulfotransferase (TPST) enzyme. The sulfated PSK can bind to its cell surface receptors, but the transcriptional readouts remain largely unknown. Here, we treated tpst mutant that are unable to produce native sulfated PSK with synthetic active PSK to capture time-series and tissue-specific readouts of PSK signaling.
Project description:Background & Aims: Non-alcoholic fatty liver disease (NALFLD)-associated changes in gut microbiota are important drivers of disease progression toward fibrosis. Therefore, reversing microbiota alterations could ameliorate NAFLD progression. Oat beta-glucan, a non-digestible polysaccharides, has shown promising therapeutic effects on hyperlipidemia associated with NAFLD, but its impact on gut microbiota and most importantly NAFLD fibrosis remains unknown. Methods: We performed detailed metabolic phenotyping including body composition, glucose tolerance, and lipid metabolism as well as comprehensive characterization of the gut-liver axis in a western-style diet (WSD)-induced model of NAFLD and assessed the effect of a beta-glucan intervention on early and advanced liver disease. Gut microbiota was modulated using broad-spectrum antibiotic (Abx) treatment. Results: Oat beta-glucan supplementation did not affect WSD-induced body weight gain, glucose intolerance, and the metabolic phenotype remained largely unaffected. Interestingly, oat beta-glucan dampened NAFLD inflammation, associated with significantly reduced monocyte-derived macrophages (MoMFs) infiltration, fibroinflammatory gene expression, and strongly reduced fibrosis development. Mechanistically, this protective effect was not mediated by changes in bile acid composition or signaling, but was dependent on gut microbiota and was lost upon Abx treatment. Specifically, oat beta-glucan partially reversed unfavorable changes in gut microbiota, resulting in an expansion of protective taxa, including Ruminococcus, and Lactobacillus followed by reduced translocation of TLR ligands. Conclusions: Our findings identify oat beta-glucan as a highly efficacious food supplement that dampens inflammation and fibrosis development in diet-induced NAFLD. These results, along with its favorable dietary profile, suggest that it may be a cost-effective and well-tolerated approach to preventing NAFLD progression and should be assessed in clinical studies.