28 research outputs found

    Probiotic properties and stress response of lactic acid bacteria isolated from cooked meat products

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    3 p.This work was supported by the Universidad Autónoma Metropolitana and the Spanish Ministry of Economy and Competitiveness (grant AGL2015-65010-C3-1-R).Peer reviewe

    Construction and validation of a mCherry protein vector for promoter analysis in Lactobacillus acidophilus

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    Lactobacilli are widespread in natural environments and are increasingly being investigated as potential health modulators. In this study, we have adapted the broad-host-range vector pNZ8048 to express the mCherry protein (pRCR) to expand the usage of the mCherry protein for analysis of gene expression in Lactobacillus. This vector is also able to replicate in Streptococcus pneumoniae and Escherichia coli. The usage of pRCR as a promoter probe was validated in Lactobacillus acidophilus by characterizing the regulation of lactacin B expression. The results show that the regulation is exerted at the transcriptional level, with lbaB gene expression being specifically induced by co-culture of the L. acidophilus bacteriocin producer and the S. thermophilus STY-31 inducer bacterium. © 2014, Society for Industrial Microbiology and Biotechnology.This study was supported by grants AGL2012-40084-C03-01, AGL2012-35814 and RM2011-00003-00-00 from the Spanish Ministry of Economics and Competitiveness and by European Commission FP7 Initial Training Network (contract 238490).Peer reviewe

    A bacteriocin gene cluster able to enhance plasmid maintenance in Lactococcus lactis

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    [Background] Lactococcus lactis is widely used as a dairy starter and has been extensively studied. Based on the acquired knowledge on its physiology and metabolism, new applications have been envisaged and there is an increasing interest of using L. lactis as a cell factory. Plasmids constitute the main toolbox for L. lactis genetic engineering and most rely on antibiotic resistant markers for plasmid selection and maintenance. In this work, we have assessed the ability of the bacteriocin Lactococcin 972 (Lcn972) gene cluster to behave as a food-grade post-segregational killing system to stabilize recombinant plasmids in L. lactis in the absence of antibiotics. Lcn972 is a non-lantibiotic bacteriocin encoded by the 11-kbp plasmid pBL1 with a potent antimicrobial activity against Lactococcus.[Results] Attempts to clone the full lcn972 operon with its own promoter (P972), the structural gene lcn972 and the immunity genes orf2-orf3 in the unstable plasmid pIL252 failed and only plasmids with a mutated promoter were recovered. Alternatively, cloning under other constitutive promoters was approached and achieved, but bacteriocin production levels were lower than those provided by pBL1. Segregational stability studies revealed that the recombinant plasmids that yielded high bacteriocin titers were maintained for at least 200 generations without antibiotic selection. In the case of expression vectors such as pTRL1, the Lcn972 gene cluster also contributed to plasmid maintenance without compromising the production of the fluorescent mCherry protein. Furthermore, unstable Lcn972 recombinant plasmids became integrated into the chromosome through the activity of insertion sequences, supporting the notion that Lcn972 does apply a strong selective pressure against susceptible cells. Despite of it, the Lcn972 gene cluster was not enough to avoid the use of antibiotics to select plasmid-bearing cells right after transformation.[Conclusions] Inserting the Lcn972 cluster into segregational unstable plasmids prevents their lost by segregation and probable could be applied as an alternative to the use of antibiotics to support safer and more sustainable biotechnological applications of genetically engineered L. lactis.This work has been funded by grant BIO2010-17414 (Ministerio de Ciencia e Innovación, Spain). Work of PL and MLM was supported by grant AGL2012-40084-C03-01 (Ministerio de Economía y Competitividad, Spain). We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). C.R. was a recipient of a predoctoral JAE-CSIC fellowship. We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)Peer reviewe

    Mídia Ninja, mídia tradicional e accountability

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    Created in 2011, the alternative mediatic group "Mídia Ninja" gained more visibility in June this year, covering and participating in marches and outdoor demonstrations in Brazil. Their streaming recordings were aired live and mostly uninterruptedly via internet, generating a large audience. As a consequence, those images and their impact ended up guiding the mainstream press and were responsible for some of the shifts of mainstream media framing about the fact. The alternativa media nourish itself from and at the same time, opposing the mainstream media. On the other hand, the mainstream media is guided by images the coverage of alternative media. Society has suspicions on the hegemonic discourse but also suspects the motives and funds of Mídia Ninja. In the midst of those issues, can we ask who has the ultimate accountability?Surgido em 2011, o grupo Mídia Ninja ganhou mais visibilidade em junho de 2013, cobrindo e participando das manifestações de rua ocorridas no Brasil neste período. Suas imagens, veiculadas ao vivo e ininterruptamente pela internet, obtiveram grande audiência, pautaram a grande imprensa e tiveram grande influência na mudança de discurso da mídia hegemônica sobre o fato. A mídia alternativa se alimentou contrapondo-se ao discurso hegemônico e a grande mídia foi pautada por imagens e coberturas da mídia alternativa, fato que fugiu do habitual. A sociedade desconfiou do discurso hegemônico, mas também dos motivos e financiamento dos “ninjas”. Em meio a essas questões, nossa intenção é refletir sobre o accountability noticioso.Surgido en 2011, el grupo "Mídia Ninja" ganó más visibilidad en junio deste año, cuando cubre y participa de las manifestaciones callejeras en Brasil. Estas imágenes fueran emitidas en directo y sin interrupciones por la Internet, han tenido amplia audiencia, han guiado a la gran prensa y fueron responsables por algunos de los cambios del discurso hegemónico de los medios sobre el hecho. Los medios de comunicación alternativa se alimentan en oposición al discurso hegemónico de los médios dominantes mientras estos son pautados por imágenes y coberturas de los medios alternativos. La sociedad sospecha del discurso hegemónico, sino también de las motivaciones y financiación del grupo Ninja. En medio de estas cuestiones, reflejamos con quien se queda la responsabilidad final

    Probiotic properties of EPS-producing Pediococcus strains

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    3 p.International Symposium on Immunonutrition 2017 Madrid, 17th–19th July, 2017 10th AnniversaryA. Pérez-Ramos was supported by the FPI grant BES-2013-065157 from the Spanish Ministry of Economy and Competitiveness. M.G. Llamas is recipient of a grant of the Basque Country Government for junior researchers in the scientific-technological environment of the fish farming and food basque sector. This work was supported by the Spanish Ministry of Economy and Competitiveness (grants AGL2012- 40084-C03 and AGL2015-65010-C3-1-R).Peer reviewe

    Análisis proteómico de Oenococcus oeni ATCC‐BAA1163 y Lactobacillus plantarum WCFS1

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    Comunicación presentadas en la 6ª reunión de la Red temática BAL (Participación de las Bacterias Lácticas en la Salud Humana y en la Calidad Alimentaria), celebrada el 28 y 29 de junio de 2012 en Tarragona.Peer reviewe

    A partial proteome reference map of the wine lactic acid bacterium Oenococcus oeni ATCC BAA-1163

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    10 p.- 5 fig.Oenococcus oeni is the main lactic acid bacterium that carries out the malolactic fermentation in virtually all red wines and in some white and sparkling wines. Oenococcus oeni possesses an array of metabolic activities that can modify the taste and aromatic properties of wine. There is, therefore, industrial interest in the proteins involved in these metabolic pathways and related transport systems of this bacterium. In this work, we report the characterization of the O. oeni ATCC BAA-1163 proteome. Total and membrane protein preparations from O. oeni were standardized and analysed by two-dimensional gel electrophoresis. Using tandem mass spectrometry, we identified 224 different spots corresponding to 152 unique proteins, which have been classified by their putative function and subjected to bioinformatics analysisThis work was supported by European Union grant FP7-2008-FOOD-211441 BIAMFOOD and by grant AGL2012-40084-C03-01 from the Spanish Ministry of Economics and CompetitivenessPeer reviewe

    Detection of riboflavin production by Lactobacillus plantarum strains during growth and its gastrointestinal survival

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    Resumen del trabajo presentado al X Workshop de la Sociedad Española de Probióticos y Prebióticos (SEPyP), celebrado en Las Palmas de Gran Canaria del 6 al 8 de febrero de 2019.The work was supported by CYTED, project 917PTE0537

    Fluorescent protein vectors for promoter analysis in lactic acid bacteria and Escherichia coli

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    29 p.- 4 tab.-2 fig.Fluorescent reporter genes are valuable tools for real-time monitoring of gene expression in living cells. In this study we describe the construction of novel promoter-probe vectors containing a synthetic mCherry fluorescent protein gene, codon-optimized for lactic acid bacteria, divergently linked, or not, to a gene encoding the S65T and F64L variant of the green fluorescent protein. The utility of the transcriptional fusion vectors was demonstrated by the cloning of a single or two divergent promoter regions and by the quantitative evaluation of fluorescence during growth of Lactococcus lactis, Enterococcus faecalis, and Escherichia coli.This work was supported by the Spanish Ministry of Science and Innovation (grants AGL2009-12998-C03-01, AGL2009- 13361-C02-02, and CSD2007-00063 Consolider Ingenio 2010 FUN-CFOOD), the Comunidad de Madrid (grant ALIBIRD P2009/AGR-1469), and the European Union VII framework program (Initial Training Network, grant no. 238490). We thank Dr. Stephen Elson for the critical reading of the manuscript. We are grateful to M. Angeles Corrales for her technical assistance.Peer Reviewe

    Vectores de fusión transcripcional para regiones promotoras uni- y bidireccionales para su uso en bacterias lácticas

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    La presente invención se refiere a una secuencia nucleotídica recombinante que comprende la secuencia del gen mrfp que codifica para una proteína autofluorescente roja optimizada para la expresión en bacterias lácticas así como a los vectores que la comprenden. La invención también se refiere a las células que comprenden la secuencia o el vector de la invención y sus usos. Así mismo, también se refiere al uso tanto de la secuencia como del vector para la detección y caracterización de una región promotora unidireccional o bidireccional, así como de su uso para transcripción. Además, se refiere al uso de las células que contienen la secuencia de la invención o el vector de la invención para la determinación de la capacidad colonizadora de células. También se refiere a los métodos relacionados con los usos de la invención, a un kit que comprende los elementos de la invención y su uso.Peer reviewedConsejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic
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