Tyrosine-containing peptides are precursors of tyramine produced by Lactobacillus plantarum strain IR BL0076 isolated from wine

Abstract Background Biogenic amines are molecules with allergenic properties. They are found in fermented products and are synthesized by lactic acid bacteria through the decarboxylation of amino acids present in the food matrix. The concentration of biogenic amines in fermented foodstuffs is influenced by many environmental factors, and in particular, biogenic amine accumulation depends on the quantity of available precursors. Enological practices which lead to an enrichment in nitrogen compounds therefore favor biogenic amine production in wine. Free amino acids are the only known precursors for the synthesis of biogenic amines, and no direct link has previously been demonstrated between …

Expression communes de gènes de M. truncatula lors de la mycorhization, la nodulation et l'interaction avec une rhizobactérie bénéfique

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Inactivation of PadR, the repressor of the phenolic acid stress response, by molecular interaction with Usp1, a universal stress protein from Lactobacillus plantarum, in Escherichia coli

ABSTRACT The phenolic acid decarboxylase gene padA is involved in the phenolic acid stress response (PASR) in gram-positive bacteria. In Lactobacillus plantarum , the padR gene encodes the negative transcriptional regulator of padA and is cotranscribed with a downstream gene, usp1 , which encodes a putative universal stress protein (USP), Usp1, of unknown function. The usp1 gene is overexpressed during the PASR. However, the role and the mechanism of action of the USPs are unknown in gram-positive bacteria. Therefore, to gain insights into the role of USPs in the PASR; (i) a usp1 deletion mutant was constructed; (ii) the two genes padR and usp1 were coexpressed with padA under its own promo…

Exopolysaccharide produced by Weissella confusa: Chemical characterisation, rheology and bioactivity

Abstract The purpose of this study was to characterise the production of exopolysaccharide (EPS) by indigenous lactic acid bacteria (LAB) isolated from traditional Algerian dairy products and to evaluate their possible use in agri-foods. Among the collection of isolated strains, the strain Weissella confusa (W4) was selected for its ability to produce EPS once exposed to a sucrose culture medium. EPS produced were first isolated with a standardised method and further characterised in terms of molecular size, antioxidant activity, and rheological properties. Its direct implication in the texture and syneresis of acid milk gel was evaluated offering interesting industrial applications for its…

Quantification of HSP27 and HSP70 Molecular Chaperone Activities

Stress-inducible heat-shock proteins (HSPs, like HSP70 and HSP27) are molecular chaperones that -protect cells from stress damage by keeping cellular proteins in a folding competent state and preventing them from irreversible aggregation. HSP27 and HSP70 chaperone activities are useful indicators to test chemical products and physical stress impact on protein denaturation, to select HSP inhibitors, or to -determine the implication of the chaperone function in other HSP activities, such as apoptosis. We have developed two simple and fast chaperone activity tests for HSP27 and HSP70 that we initially set up to test the effect of potential HSP inhibitors obtained after screening of chemical an…

Proteomics: a functional genomic approach for studying mycorrhizal interactions in legumes

A technical trick for studying proteomics in parallel to transcriptomics in symbiotic root-fungus interactions

We have developed a protocol in which proteins and mRNA can be analyzed from single root samples. This experimental design was validated in arbuscular mycorrhiza by comparing the proteins profiles obtained with those from a classical protein extraction process. It is a step forward to make simultaneous proteome and transcriptiome profiling possible.

Differential expression of the agr locus of Listeria monocytogenes EGD-e

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Signal-related gene responses in beneficial root interactions with non-rhizobial microorganisms

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Agr system of Listeria monocytogenes EGD-e: role in adherence and differential expression pattern.

ABSTRACT In this study, we investigated the agrBDCA operon in the pathogenic bacterium Listeria monocytogenes EGD-e. In-frame deletion of agrA and agrD resulted in an altered adherence and biofilm formation on abiotic surfaces, suggesting the involvement of the agr system of L. monocytogenes during the early stages of biofilm formation. Real-time PCR experiments indicated that the transcript levels of agrBDCA depended on the stage of biofilm development, since the levels were lower after the initial attachment period than during biofilm growth, whereas transcription during planktonic growth was not growth phase dependent. The mRNA quantification data also suggested that the agr system was a…

Gènes activés lors de l'interaction racine / Glomus mosseae chez Medicago truncatula

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Protective role of glutathione addition against wine-related stress in Oenococcus oeni

FIliació URV: SIInclòs a la memòria: SI Oenococcus oeni is the main species responsible for the malolactic fermentation (MLF) of wine due to its ability to survive in this environment. Some wine-related stress factors, such as ethanol and low pH, may alter the cell redox balance of O. oeni. For the first time, the ability to uptake glutathione (GSH), an almost universal tripeptide with antioxidant properties, has been associated to the improvement of stress response in O. oeni. Despite the inability of O. oeni to synthesize GSH, this bacterium can capture it from the media. The ability of 30 O. oeni strains to uptake GSH was assessed in this study. Although all of the strains tested were ab…

Inactivation of a small heat shock protein affects cell morphology and membrane fluidity in Lactobacillus plantarum WCFS1.

A small heat shock gene of Lactobacillus plantarum strain WCFS1 was deleted using a Cre-lox based system. Compared to the wild type, the ∆hsp 18.55 mutant strain displayed a similar growth rate when cultivated either under optimal temperature or under different stress conditions such as heat, low pH and salt stress. However, a longer lag phase was observed when the ∆hsp 18.55 mutant strain was cultivated under short intense heat stress (50 °C). This suggests that the hsp 18.55 gene of L. plantarum may be involved in recovery of L. plantarum stressed cells in the early stage of high temperature stress. In addition, morphology of the mutant cells, investigated by scanning electron microscopy,…

Chemical Transfers Occurring Through Oenococcus oeni Biofilm in Different Enological Conditions

International audience; Chardonnay wine malolactic fermentations were carried out to evaluate the chemical transfers occurring at the wood/wine interface in the presence of two different bacterial lifestyles. To do this, Oenococcus oeni was inoculated into must and wine in its planktonic and biofilm lifestyles, whether adhering or not to oak chips, leading to three distinct enological conditions: (i) post-alcoholic fermentation inoculation in wine in the absence of oak chips, (ii) post-alcoholic fermentation inoculation in wine in the presence of oak chips, and (iii) co-inoculation of both Saccharomyces cerevisiae and O. oeni directly in Chardonnay musts in the presence of oak chips. Classi…

Distinct amino acids of  the Oenococcus oeni small heat shock protein Lo18 are essential for damaged protein protection and membrane stabilization

The small heat shock protein (smHsp) Lo18 from lactic acid bacteria Oenococcus oeni reduces in vitro thermal aggregation of proteins and modulates the membrane fluidity of native liposomes. An absence of information relating to the way in which the smHsp demonstrates a stabilizing effect for both proteins and membranes prompted this study. We expressed three Lo18 proteins with amino acid substitutions in Escherichia coli to investigate their ability to prevent E. coli protein aggregation and their capacity to stabilize E. coli whole-cell membranes. Our results showed that the alanine 123 to serine substitution induces a decrease in chaperone activity in denaturated proteins, and that the ty…

Gene responses to non rhizobial beneficial microbe interactions in legumes

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Technological properties of Oenococcus oeni strains isolated from typical southern Italian wines.

Aims:  To isolate indigenous Oenococcus oeni strains suitable as starters for malolactic fermentation (MLF), using a reliable polyphasic approach. Methods and Results: Oenococcus oeni strains were isolated from Nero di Troia wines undergoing spontaneous MLF. Samples were taken at the end of alcoholic fermentation and during MLF. Wine samples were diluted in a sterile physiological solution and plated on MRS and on modified FT80. Identification of O. oeni strains was performed by a polymerase chain reaction (PCR) experiment using strain-specific primers. Strains were further grouped using a multiplex RAPD-PCR analysis. Then, six strains were inoculated in two wine-like media with two differe…

Development of  a qPCR assay for specific quantification of Botrytis cinerea on grapes

The aim of this study was to develop a system for rapid and accurate real-time quantitative PCR (qPCR) identification and quantification of Botrytis cinerea , one of the major pathogens present on grapes. The intergenic spacer (IGS) region of the nuclear ribosomal DNA was used to specifically detect and quantify B. cinerea . A standard curve was established to quantify this fungus. The qPCR reaction was based on the simultaneous detection of a specific IGS sequence and also contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes that inhibit PCR. In these conditions, the assay had high efficiency (97%), and the limit o…

Characterization of the CtsR stress response regulon in Lactobacillus plantarum.

ABSTRACT Lactobacillus plantarum ctsR was characterized. ctsR was found to be cotranscribed with clpC and induced in response to various abiotic stresses. ctsR deletion conferred a heat-sensitive phenotype with peculiar cell morphological features. The transcriptional pattern of putative CtsR regulon genes was examined in the Δ ctsR mutant. Direct CtsR-dependent regulation was demonstrated by DNA-binding assays using recombinant CtsR and the promoters of the ctsR - clpC operon and hsp1 .