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Genome wide transcription start sites analysis of the Xanthomonas campestris pv. campestris B100 with insights into the gum gene cluster directing the biosynthesis of the exopolysaccharide xanthan

ABSTRACT: Genome wide transcription start sites analysis of the Xanthomonas campestris pv. campestris B100 with insights into the gum gene cluster directing the biosynthesis of the exopolysaccharide xanthan

PROVIDER: PRJEB12463 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR1224293.fastq.gz	Fastqsanger.gz

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Strength of a “single-tube” protocol for protein isolation from multiple cell fractions shown for the plant pathogen Xanthomonas campestris pv. campestris B100 with a focus on proteins involved in the biosynthesis of the exopolysaccharide xanthan

Project description:Xanthomonas campestris pv. campestris (Xcc) B100 is a phytopathogenic bacterium, which uses xanthan, a secreted heterogeneous exopolysaccharide in both stages of infection of Brassicaceae. Until now, proteome analysis of Xcc lacking a detailed view on the proteins involved in xanthan biosynthesis. Proteins involved in the biosynthesis of this polysaccharide are located near, in or at the cell membrane. This study aims to establish a robust and rapid protocol for a comprehensive proteome analysis of Xcc strains, without the need to isolate different cell fractions. Therefore, a method for the analysis of the whole cell proteome was compared to the isolation of specific fractions regarding the total number of identified proteins, the overlap as well as the differences between the approaches. The whole cell proteome analysis using the “single-tube” preparation protocol resulted in more than 2,000 identified proteins with a coverage of 95.11% of the cell fraction specific identifications. To prove the usability of the protocol to study natural product formation at the envelope with proteins distributed over various cell fractions, Gum proteins of Xcc B100 were identified and quantified by a label free quantification (LFQ) approach. Thus, expression profiles of 9 out of 12 Gum proteins comparing stationary phase and growth phase were measured and showed differential expression levels within the operon structure. Five out of ten identified Gum proteins show a differential regulation indicating regulatory mechanisms beyond transcription level.

2022-02-16 | PXD027261 | Pride

Xanthomonas campestris pv. campestris str. B100

Project description:Xanthomonas campestris pv. campestris B100 Transcriptomes

| PRJNA436896 | ENA

Comparative microarrays of Xanthomonas campestris pv. camp. B100 Δcrt1 with the Xanthomonas camp. pv camp B100

Project description:To investigate the effect of the transcriptional regulator Crt1 on the transcirptome of Xanthomonas campestris pv. camp (Xcc), comparative genome-wide transcriptome analysis was conducted. For this purpose, the wild-type strain Xcc B100 and the mutant strain Xcc Δcrt1 were each cultivated in triplicates in minimal medium supplemented with glucose as sole carbon source. RNA samples from the biological replicates were obtained at an early stationary growth stage. RNA was isolated and the three replicates were combined for each strain. Furthermore, the data from two arrays (dye swap) were combined to provide statistically reliable conclusions.

2022-10-25 | E-MTAB-12383 | biostudies-arrayexpress

Widespread consumption of the common food additive xanthan gum by the human gut microbiota

Project description:The diets of industrialized countries reflect the increasing use of processed foods, often with the inclusion of novel food additives. Xanthan gum is a complex polysaccharide with unique rheological properties that have established its use as a widespread stabilizer and thickening agent. Xanthan gum’s chemical structure is distinct from the host and dietary polysaccharides that are more commonly expected to transit the gastrointestinal tract, and little is known about its direct interaction with the gut microbiota, which plays a central role in digestion of other dietary fiber polysaccharides. Here, we show that the ability to digest xanthan gum is surprisingly common in industrialized human gut microbiomes and appears contingent on a single uncultured bacterium in the family Ruminococcaceae. Our data reveal that this primary degrader cleaves the xanthan gum backbone before processing the released oligosaccharides using additional enzymes. Surprisingly, some individuals harbor a Bacteroides intestinalis that is incapable of consuming polymeric xanthan gum but grows on oligosaccharide products generated by the Ruminococcaceae. Feeding xanthan gum to germfree mice colonized with a human microbiota containing the uncultured Ruminococcaceae supports the idea that this additive can drive expansion of this primary degrader along with exogenously introduced Bacteroides intestinalis. Our work demonstrates the existence of a potential xanthan gum food chain involving at least two members of different phyla of gut bacteria and provides an initial framework to understand how widespread consumption of a recently introduced food additive influences human microbiomes.

2022-02-28 | PXD031522 | Pride

Comprehensive proteome profiling of a Xanthomonas campestris pv campestris B100 culture grown in a minimal medium with a specific focus on nutrient consumption and xanthan biosynthesis

Project description:Xanthan is the most used polysaccharide of bacterial origin. There are many industrial applications for xanthan, especially in food products. The organism Xanthomonas campestris is commonly used for xanthan fermentation, but little is known about the xanthan synthesis mechanism on the proteome level. The study used a label-free LC-MS/MS method to investigate the protein shifts during a typical xanthan fermentation of 72 hours. Two thousand four hundred sixteen proteins were identified, representing 54.75 % of the 4413 entries in the reference database. One thousand seven hundred forty-nine proteins were measured in a quantifiable quality. The quality of the proteome data was examined by the abundance changes of the ribosomal proteins according to the growth phases. The phenotypic changes during the cultivation, especially processes known to be influencing or close related to xanthan production, were investigated by proteome analysis. These processes were nitrogen metabolism, sugar nucleotide metabolism, and xanthan synthesis. This study shows the abundance of changes in central metabolites during a xanthan fermentation of Xcc. The presented research provides crucial information for further biotechnological studies to improve xanthan production.

2024-05-24 | PXD041470 | Pride

Promoter analysis of the Xanthomonas campestris pv. campestris gum operon directing biosynthesis of the xanthan polysaccharide.

Project description:The Xanthomonas campestris gum gene cluster is composed of 12 genes designated gumB, -C, -D, -E, -F, -G, -H, -I, -J, -K, -L, and -M. The transcriptional organization of this gene cluster was analyzed by the construction of gum-lacZ transcriptional fusions in association with plasmid integration mutagenesis. This analysis, coupled with primer extension assays, indicated that the gum region was mainly expressed as an operon from a promoter located upstream of the first gene, gumB.

| S-EPMC178194 | biostudies-other

RNAseq-based reannotation of the complete genome of Xcc B100

Project description:Refined annotation of the complete genome of the phytopathogenic and xanthan producing Xanthomonas campestris pv. campestris strain B100 based on RNA sequence data

| PRJEB19915 | ENA

Transcriptomics of Xanthomonas campestris pv campestris in cabbage xylem sap

Project description:Xylem sap of young cabbage plantlets was recovered from root pressure exudation and used as a growth medium for the vascular pathogen Xanthomonas campestris pv campestris, the causative agent of the black rot of Brassicaceae.

2014-01-04 | E-MTAB-2290 | biostudies-arrayexpress

Transcriptomic response of Xanthomonas campestris pv. campestris after exposure to allyl-isothiocyanate and indol-3 carbinol

Project description:We performed a transcriptomic analysis of the necrotrophic bacteria Xanthomonas campestris pv. campestris exposed to two different isothiocyanates (allyl-isothiocyanate and indol-3-carbinol), searching for mechanisms of adaptation and detoxification of these chemicals.

2021-09-08 | GSE167555 | GEO

Early transcriptional response to phytopathogen Xanthomonas campestris pv. campestris by susceptible cabbage host

Project description:Black rot, caused by Xanthomonas campestris pv. campestris (Xcc) is one of the most devastating diseases of cruciferous crops worldwide. The pathogen infects and multiplies in plant vascular tissues and, as the disease progresses, the veins of infected tissues turn black and characteristic V-shaped lesions appear along the margins of leaves.The aim of this work is to identify differentially expressed genes from Brassica oleracea during early infection by Xcc, in an attempt to identify proteins related to resistance. Cabbge seedlings were inoculated with Xanthomonas campestris pv campestris (Xcc) suspension and cabbage gene expression at 6h., 24h. And 48h. After inoculation was assessed with help of Brassica 95k EST microarray chip.

2015-05-09 | E-GEOD-68670 | biostudies-arrayexpress

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