Nueva enzima para la biosíntesis de isoprenoides.

Se ha encontrado una nueva enzima con actividad 1-desoxi-D-xilulosa 5-fosfato reductoisomerasa que cataliza la reacción de producción de 2-C-metil-D-eritritol 4-fosfato a partir de 1-desoxi-D-xilulosa 5-fosfato, que tiene la secuencia de aminoácidos SEQ ID NO: 41. La enzima es útil en la síntesis de isoprenoides, particularmente en la síntesis de 2-C-metil-D-eritritol 4-fosfato.Solicitud: 201031068 (14.07.2010)Nº Pub. de Solicitud: ES2372942A1 (30.01.2012)Nº de Patente: ES2372942B1 (04.12.2012

Evaluation of the effects of erythritol on gene expression in Brucella abortus

Bacteria of the genus Brucella have the unusual capability to catabolize erythritol and this property has been associated with their virulence mainly because of the presence of erythritol in bovine foetal tissues and because the attenuated S19 vaccine strain is the only Brucella strain unable to oxydize erythritol. In this work we have analyzed the transcriptional changes produced in Brucella by erythritol by means of two high throughput approaches: RNA hybridization against a microarray containing most of Brucella ORF's constructed from the Brucella ORFeome and next generation sequencing of Brucella mRNA in an Illumina GAIIx platform. The results obtained showed the overexpression of a group of genes, many of them in a single cluster around the ery operon, able to co-ordinately mediate the transport and degradation of erythritol into three carbon atoms intermediates that will be then converted into fructose-6P (F6P) by gluconeogenesis. Other induced genes participating in the nonoxidative branch of the pentose phosphate shunt and the TCA may collaborate with the ery genes to conform an efficient degradation of sugars by this route. On the other hand, several routes of amino acid and nucleotide biosynthesis are up-regulated whilst amino acid transport and catabolism genes are down-regulated. These results corroborate previous descriptions indicating that in the presence of erythritol, this sugar was used preferentially over other compounds and provides a neat explanation of the the reported stimulation of growth induced by erythritol

Epitopes for Multivalent Vaccines Against Listeria, Mycobacterium and Streptococcus spp: A Novel Role for Glyceraldehyde-3-Phosphate Dehydrogenase

The glycolytic enzyme and bacterial virulence factor of Listeria monocytogenes, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Lmo2459), ADP-ribosylated the small GTPase, Rab5a, and blocked phagosome maturation. This inhibitory activity localized within the NAD binding domain of GAPDH at the N-terminal 1?22 peptides, also conferred listeriosis protection when used in dendritic cell-based vaccines. In this study, we explore GAPDH of Listeria, Mycobacterium, and Streptococcus spp. taxonomic groups to search for epitopes that confer broad protection against pathogenic strains of these bacteria. GAPDH multivalent epitopes are selected if they induce inhibitory actions and wide-ranging immune responses. Proteomic isolation of GAPDH from dendritic cells infected with Listeria, Mycobacterium, or Streptococcus confirmed similar enzymatic, Rab5a inhibitory and immune stimulation abilities. We identified by bioinformatics and functional analyses GAPDH N-terminal 1?22 peptides from Listeria, Mycobacterium, and Streptococcus that shared 95% sequence homology, enzymatic activity, and B and T cell immune domains. Sera obtained from patients or mice infected with hypervirulent pathogenic Listeria, Mycobacterium, or Streptococcus presented high levels of anti-GAPDH 1?22 antibodies and Th2 cytokines. Monocyte derived dendritic cells from healthy donors loaded with GAPDH 1?22 peptides from Listeria, Mycobacterium, or Streptococcus showed activation patterns that correspond to cross-immunity abilities. In summary, GAPDH 1?22 peptides appeared as putative candidates to include in multivalent dendritic based vaccine platforms for Listeria, Mycobacterium, or Streptococcus

Dispositivo de desinfección de productos mediante ozono

Dispositivo de desinfección de productos mediante ozono; comprendiendo un aparato portátil de generación de ozono, mediante la disociación de las moléculas de oxígeno contenidas en el aire, provisto de un ventilador para el suministro del ozono a través de una boca de salida. Dicho dispositivo comprende un contenedor desechable, provisto de una boca de entrada adecuada para el acoplamiento del aparato generador de ozono; delimitando dicho contenedor al menos una cámara de desinfección, de volumen variable, en comunicación dicha cámara con la mencionada boca de salida del aparato de generación de ozono, y que comprende unos medios de ajuste del volumen de la cámara al tamaño del producto o productos a desinfectar.Solicitud: 202030375 (30.04.2020)Nº Pub. de Solicitud: ES2873352A1 (03.11.2021

Generation of the Brucella melitensis ORFeome version 1.1.

The bacteria of the Brucella genus are responsible for a worldwide zoonosis called brucellosis. They belong to the alpha-proteobacteria group, as many other bacteria that live in close association with a eukaryotic host. Importantly, the Brucellae are mainly intracellular pathogens, and the molecular mechanisms of their virulence are still poorly understood. Using the complete genome sequence of Brucella melitensis, we generated a database of protein-coding open reading frames (ORFs) and constructed an ORFeome library of 3091 Gateway Entry clones, each containing a defined ORF. This first version of the Brucella ORFeome (v1.1) provides the coding sequences in a user-friendly format amenable to high-throughput functional genomic and proteomic experiments, as the ORFs are conveniently transferable from the Entry clones to various Expression vectors by recombinational cloning. The cloning of the Brucella ORFeome v1.1 should help to provide a better understanding of the molecular mechanisms of virulence, including the identification of bacterial protein-protein interactions, but also interactions between bacterial effectors and their host's targets

Método de diagnóstico molecular para la detección de Brucella abortus y en diferenciación de la cepa vacunal B19.

Se describen una serie de métodos de diagnóstico molecular que permiten a diferenciación entre la estirpe vacunal Brucella abortus B19 y todas las demás estirpes del género Brucella. En un caso se aplica la amplificación de genoma, usando como iniciadores oligonucleótidos derivados de la región que contiene los genes codificantes de la ruta de utilización del eritritol. Otro grupo de métodos se basa en la hibridación de ADN genómico usando como sondas oligonucleótidos de la mencionada región eri. En este caso pueden aplicarse según la técnica denominada “hibridación tipo Southern" o alternativamente por el procedimiento del “Dot blot". Finalmente se aplican estos métodos para la identificación de gérmenes del género Brucella en cualquier muestra biológica. Los nuevos procedimientos son ventajosos sobre los métodos tradicionales, inmunológicos o de cultivo, aportando mayor especificidad y rapidez.Solicitud: 9302728 (16.12.1993)Nº Pub. de Solicitud: ES2078174A1 (01.12.1995)Nº de Patente: ES2078174B1 (16.08.1996

Evaluation of the effects of erythritol on gene expression in Brucella abortus

Bacteria of the genus Brucella have the unusual capability to catabolize erythritol and this property has been associated with their virulence mainly because of the presence of erythritol in bovine foetal tissues and because the attenuated S19 vaccine strain is the only Brucella strain unable to oxydize erythritol. In this work we have analyzed the transcriptional changes produced in Brucella by erythritol by means of two high throughput approaches: RNA hybridization against a microarray containing most of Brucella ORF's constructed from the Brucella ORFeome and next generation sequencing of Brucella mRNA in an Illumina GAIIx platform. The results obtained showed the overexpression of a group of genes, many of them in a single cluster around the ery operon, able to co-ordinately mediate the transport and degradation of erythritol into three carbon atoms intermediates that will be then converted into fructose-6P (F6P) by gluconeogenesis. Other induced genes participating in the nonoxidative branch of the pentose phosphate shunt and the TCA may collaborate with the ery genes to conform an efficient degradation of sugars by this route. On the other hand, several routes of amino acid and nucleotide biosynthesis are up-regulated whilst amino acid transport and catabolism genes are down-regulated. These results corroborate previous descriptions indicating that in the presence of erythritol, this sugar was used preferentially over other compounds and provides a neat explanation of the the reported stimulation of growth induced by erythritol

Transcriptome analysis of the Brucella abortus BvrR/BvrS two-component regulatory system

Background: The two-component BvrR/BvrS system is essential for Brucella abortus virulence. It was shown previously that its dysfunction alters the expression of some major outer membrane proteins and the pattern of lipid A acylation. To determine the genes regulated by BvrR/BvrS, we performed a whole-genome microarray analysis using B. abortus RNA obtained from wild type and bvrR mutant cells grown in the same conditions. Methodology/principal Findings: A total of 127 differentially expressed genes were found: 83 were over expressed and 44 were less expressed in the bvrR mutant. Two operons, the phosphotransferase system and the maltose transport system, were down-regulated. Several genes involved in cell envelope or outer membrane biogenesis were differentially expressed: genes for outer membrane proteins (omp25a, omp25d), lipoproteins, LPS and fatty acid biosynthesis, stress response proteins, chaperones, flagellar genes, and twelve genes encoding ABC transport systems. Ten genes related with carbon metabolism (pckA and fumB among others) were up-regulated in the bvrR mutant, and denitrification genes (nirK, norC and nosZ) were also regulated. Notably, seven transcriptional regulators were affected, including VjbR, ExoR and OmpR that were less expressed in the bvrR mutant. Finally, the expression of eleven genes which have been previously related with Brucella virulence was also altered. Conclusions/significance: All these data corroborate the impact of BvrR/BvrS on cell envelope modulation, confirm that this system controls the carbon and nitrogen metabolism, and suggest a cross-talk among some regulators to adjust the Brucella physiology to the shift expected to occur during the transit from the extracellular to the intracellular niche

Real-time PCR confirmation of selected ORFs with significant changes in mRNA expression under erythritol treatment.

<p>Real-time PCR confirmation of selected ORFs with significant changes in mRNA expression under erythritol treatment.</p

Histogram showing the distribution of regulated genes in COG categories.

<p>Bar height is proportional to the number of genes in each category, each vertical mark representing ten genes. Bars above and below the axis represent up- and down-regulated genes, respectively. Categories containing 5 or less regulated genes in each of the experimental approaches have been omitted from the histogram.</p