- Email: [email protected]
Study of Oenococcus oeni gene expression to improve the quality of wines
Invited speaker texts / Current Opinion in Biotechnology 24S (2013) S21–S27
RLO. Additionally, it was possible to identify the infected organ for each fish: be it the spleen, skin or the liver. This information represents an important and useful tool to improve our knowledge of RMS and our understanding of the route of infection, which has not yet been confirmed. http://dx.doi.org/10.1016/j.copbio.2013.05.027 Antibody based systems for the determination of mycotoxins in food and feed Ozlem Ertekin 1 , Fatma Betul Guloglu 1 , Seyda Pirincci 1 , Sevecen Tuglu 1 , Esin Akcael 1 , Duygu Ercan 1 , Ibrahim Hatipoglu 1 , Meltem Goksel 2 , Vefa Ahsen 2 , Selma Ozturk 1 1 TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, GebzeKocaeli, Turkey 2 Gebze Institute of High Technology, Faculty of Science, Department of Chemistry, Gebze, Kocaeli, Turkey E-mail address: [email protected] (S. Ozturk).
Food and feed security is a crucial component of public health issues and several directives are employed in order to prevent health risks associated with their contamination. In many cases, the same product is required to be tested for several risk factors, which in turn leads to increased cost and time due to difficult and complicated analysis. These analyses are mostly laboratory based tests which require trained staff and expensive equipment. Analyte specific antibodies are widely used as a part of detection systems. Depending on the specificity and affinity of the antigen–antibody interaction, immune-diagnosis serves a variety of options for efficient analyte detection and is amongst the most widely used systems. In the context of ongoing projects, we are working on the antibody based systems for the detection of several mycotoxins such as Aflatoxin B1, M1, Ochratoxin, Patulin and Zeraleanon in different food and feed samples. In order to be able to produce mycotoxin specific antibodies, we initially conjugated the toxins to an immunogenic carrier protein and injected to test animals. After monoclonal antibody production, the antibodies were characterized and modified to be used in ELISA test systems and immunoaffinity columns. Multiple mycotoxin extraction studies were done by food engineers and developed test systems have been used for multiplexed mycotoxin analysis in the extracts containing more than one toxin. Acknowledgements: This work was supported by T.C. Ministry of Food, Agricultural and Foresty (Project no: TAGEM/10/AR-GE/10) and TUBITAK (Project no: 110G139) http://dx.doi.org/10.1016/j.copbio.2013.05.028 Study of Oenococcus oeni gene expression to improve the quality of wines Lucilla Iacumin, Marco Vendrame, Giuseppe Comi, Marisa Manzano Dipartimento di ScienzedegliAlimenti, Università di Udine, via Sondrio 2/A, 33100 Udine, Italy E-mail address: [email protected] (L. Iacumin). Among LAB, Oenococcus oeni is without any doubt the best-adapted species to the wine environment, and is often used as a starter to perform malolactic fermentation (MLF) and ensure the successful completion of MLF as well as to encourage a positive flavour development. The expression of mleA gene, which encodes for the malolactic enzyme in O. oeni, is a crucial point in MLF. Besides the capacity of performing malolactic fermentation, LAB are capable to
S23
influence the aromatic complexity of wine, thanks to the release of volatile compounds due to the activity of the ˇ-glucosidase enzyme, which has been isolated in various strains, including O. oeni. For this reasons, the purpose of the study was to evaluate the influence of different environmental conditions, such as l-malic and l-lactic acid, ethanol, co-inoculation with Saccharomyces cerevisiae, sequential inoculum on the expression of the mleA gene and the gene encoding for the ˇ-glucosidase enzyme. The results obtained show that co-inoculation with Saccharomyces cerevisiae and high concentrations of ethanol in the medium are the best conditions to ensure high levels of mleA gene transcription. The same conclusion was obtained studying the gene expression for the ˇ-glucosidase enzyme. In fact, results point out that, during co-inoculation, higher levels of expression are registered. Therefore, and although winemakers try often to avoid this practice, co-inoculation can be considered the best winemaking scenario to ensure both rapid completion of MLF and expression of ˇglucosidase encoding gene, which can lead to the release of positive aromatic volatile compounds in wine. http://dx.doi.org/10.1016/j.copbio.2013.05.029 Free oligosaccharides accumulation and tissue damages in Ma2c1 deficient mice Tommaso Beccari 1 , Paciotti Silvia 1 , Persichetti Emanuele 1 , Codini Michela 1 , Tasegian Anna 1 , Gieselmann Volkmar 2 , Dieter Hartmann 2 , Duvet Sandrine 3 1
DSEEA, University of Perugia, Perugia, Italy University of Bonn, Bonn, Germany 3 Unité de Glycobiologie Structurale et Fonctionnelle UMR/CNRS, France E-mail address: [email protected] (T. Beccari). 2
Free Man9-8GlcNAc2 are released during the biosynthesis pathway of N-linked glycans or from misfolded glycoproteins during the ERAD-process and are reduced to Man5GlcNAc in the cytosol. In this form free oligosaccharides can be transferred into the lysosomes to be completely degraded. Alpha-mannosidase (MAN2C1) is the enzyme responsible for the partial demannosylation occurring in free oligosaccharides in the cytosol. Studies performed to better understand the role of this enzyme demonstrated that the inhibition of MAN2C1 expression induces accumulation of Man9-8GlcNAc oligosaccharides and apoptosis in vitro. We investigated the consequences caused by the lack of cytosolic alpha-mannosidase activity in vivo by the generation of Man2c1 deficient mice. Accumulation of Man9-8GlcNAc oligosaccharides was recognized in all analyzed tissues. Histological analysis revealed neuronal and glial degeneration with formation of multiple vacuoles in layer V and white matter. Large clear vacuoles in retronuclear position were also detected in small intestine enterocytes. In liver multiple droplets filled with lipids and bile constituents along with vacuolization were observed. In addition lectin screening showed the presence of saccharides in microvascular endothelia of Man2c1 deficient tissues. These findings confirm the key role played by Man2c1 in the catabolism of free oligosaccharide suggesting that its deficiency could cause a not yet described disease in humans. http://dx.doi.org/10.1016/j.copbio.2013.05.030
RLO. Additionally, it was possible to identify the infected organ for each fish: be it the spleen, skin or the liver. This information represents an important and useful tool to improve our knowledge of RMS and our understanding of the route of infection, which has not yet been confirmed. http://dx.doi.org/10.1016/j.copbio.2013.05.027 Antibody based systems for the determination of mycotoxins in food and feed Ozlem Ertekin 1 , Fatma Betul Guloglu 1 , Seyda Pirincci 1 , Sevecen Tuglu 1 , Esin Akcael 1 , Duygu Ercan 1 , Ibrahim Hatipoglu 1 , Meltem Goksel 2 , Vefa Ahsen 2 , Selma Ozturk 1 1 TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute, GebzeKocaeli, Turkey 2 Gebze Institute of High Technology, Faculty of Science, Department of Chemistry, Gebze, Kocaeli, Turkey E-mail address: [email protected] (S. Ozturk).
Food and feed security is a crucial component of public health issues and several directives are employed in order to prevent health risks associated with their contamination. In many cases, the same product is required to be tested for several risk factors, which in turn leads to increased cost and time due to difficult and complicated analysis. These analyses are mostly laboratory based tests which require trained staff and expensive equipment. Analyte specific antibodies are widely used as a part of detection systems. Depending on the specificity and affinity of the antigen–antibody interaction, immune-diagnosis serves a variety of options for efficient analyte detection and is amongst the most widely used systems. In the context of ongoing projects, we are working on the antibody based systems for the detection of several mycotoxins such as Aflatoxin B1, M1, Ochratoxin, Patulin and Zeraleanon in different food and feed samples. In order to be able to produce mycotoxin specific antibodies, we initially conjugated the toxins to an immunogenic carrier protein and injected to test animals. After monoclonal antibody production, the antibodies were characterized and modified to be used in ELISA test systems and immunoaffinity columns. Multiple mycotoxin extraction studies were done by food engineers and developed test systems have been used for multiplexed mycotoxin analysis in the extracts containing more than one toxin. Acknowledgements: This work was supported by T.C. Ministry of Food, Agricultural and Foresty (Project no: TAGEM/10/AR-GE/10) and TUBITAK (Project no: 110G139) http://dx.doi.org/10.1016/j.copbio.2013.05.028 Study of Oenococcus oeni gene expression to improve the quality of wines Lucilla Iacumin, Marco Vendrame, Giuseppe Comi, Marisa Manzano Dipartimento di ScienzedegliAlimenti, Università di Udine, via Sondrio 2/A, 33100 Udine, Italy E-mail address: [email protected] (L. Iacumin). Among LAB, Oenococcus oeni is without any doubt the best-adapted species to the wine environment, and is often used as a starter to perform malolactic fermentation (MLF) and ensure the successful completion of MLF as well as to encourage a positive flavour development. The expression of mleA gene, which encodes for the malolactic enzyme in O. oeni, is a crucial point in MLF. Besides the capacity of performing malolactic fermentation, LAB are capable to
S23
influence the aromatic complexity of wine, thanks to the release of volatile compounds due to the activity of the ˇ-glucosidase enzyme, which has been isolated in various strains, including O. oeni. For this reasons, the purpose of the study was to evaluate the influence of different environmental conditions, such as l-malic and l-lactic acid, ethanol, co-inoculation with Saccharomyces cerevisiae, sequential inoculum on the expression of the mleA gene and the gene encoding for the ˇ-glucosidase enzyme. The results obtained show that co-inoculation with Saccharomyces cerevisiae and high concentrations of ethanol in the medium are the best conditions to ensure high levels of mleA gene transcription. The same conclusion was obtained studying the gene expression for the ˇ-glucosidase enzyme. In fact, results point out that, during co-inoculation, higher levels of expression are registered. Therefore, and although winemakers try often to avoid this practice, co-inoculation can be considered the best winemaking scenario to ensure both rapid completion of MLF and expression of ˇglucosidase encoding gene, which can lead to the release of positive aromatic volatile compounds in wine. http://dx.doi.org/10.1016/j.copbio.2013.05.029 Free oligosaccharides accumulation and tissue damages in Ma2c1 deficient mice Tommaso Beccari 1 , Paciotti Silvia 1 , Persichetti Emanuele 1 , Codini Michela 1 , Tasegian Anna 1 , Gieselmann Volkmar 2 , Dieter Hartmann 2 , Duvet Sandrine 3 1
DSEEA, University of Perugia, Perugia, Italy University of Bonn, Bonn, Germany 3 Unité de Glycobiologie Structurale et Fonctionnelle UMR/CNRS, France E-mail address: [email protected] (T. Beccari). 2
Free Man9-8GlcNAc2 are released during the biosynthesis pathway of N-linked glycans or from misfolded glycoproteins during the ERAD-process and are reduced to Man5GlcNAc in the cytosol. In this form free oligosaccharides can be transferred into the lysosomes to be completely degraded. Alpha-mannosidase (MAN2C1) is the enzyme responsible for the partial demannosylation occurring in free oligosaccharides in the cytosol. Studies performed to better understand the role of this enzyme demonstrated that the inhibition of MAN2C1 expression induces accumulation of Man9-8GlcNAc oligosaccharides and apoptosis in vitro. We investigated the consequences caused by the lack of cytosolic alpha-mannosidase activity in vivo by the generation of Man2c1 deficient mice. Accumulation of Man9-8GlcNAc oligosaccharides was recognized in all analyzed tissues. Histological analysis revealed neuronal and glial degeneration with formation of multiple vacuoles in layer V and white matter. Large clear vacuoles in retronuclear position were also detected in small intestine enterocytes. In liver multiple droplets filled with lipids and bile constituents along with vacuolization were observed. In addition lectin screening showed the presence of saccharides in microvascular endothelia of Man2c1 deficient tissues. These findings confirm the key role played by Man2c1 in the catabolism of free oligosaccharide suggesting that its deficiency could cause a not yet described disease in humans. http://dx.doi.org/10.1016/j.copbio.2013.05.030