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The use of food-grade microorganisms such as lactic acid bacteria (LAB) is one of the most promising methods for delivering health promoting compounds. Since it is not always possible to obtain strains that have the ability to produce specific compounds naturally or that produce them in sufficient quantities to obtain physiological responses, genetic modifications can be performed to improve their output. The objective of this study was to evaluate if previously studied genetically modified LAB (GM-LAB), with proven in vivo beneficial effects, are just as safe as the progenitor strain from which they were derived. Mice received an elevated concentration of different GM-LAB or the native parental strain from which they were derived during a prolonged period of time, and different health parameters were evaluated. Similar growth rates, hematological values, and other physiological parameters were obtained in the animals that received the GM-LAB compared to those that were fed with the native strain. These results demonstrate that the GM-LAB used in this study are just as safe as the native strains from which they were derived and thus merit further studies to include them into the food chain. Fil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: van Sinderen, Douwe. University College Cork; Irlanda Fil: Hugenholtz, Jeroen. Nizo Food Research, Ede,; Países Bajos Fil: Piard, Jean Christophe. Institut National de la Recherche Agronomique; Francia Fil: Sesma, Fernando Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina

α-galactooligosaccharides (α-GOS) found in legumes such as soybeans can cause gastrointestinal disorders since mammals lack α-galactosidase (α-Gal) in the small intestine which is necessary for their hydrolysis. Lactobacillus fermentum CRL 722 is a lactic acid bacterium (LAB) capable of degrading α-GOS due to its elevated α-Gal activity. When conventional rats were fed live L. fermentum CRL 722 or cell-free extracts of this strain, a short-lived α-Gal activity was detected in the upper gastrointestinal tract. The safety of this LAB was also assessed. L. fermentum CRL 722 could thus be used as a vehicle to safely confer α-Gal in the small intestine of monogastric animal. Fil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Piard, Jean-Christophe. Institut National de la Recherche Agronomique; Francia Fil: Sesma, Fernando Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina

Alpha-galactosides are abundant sugars in legumes such as soy. Because of the lack of alpha-galactosidase (alpha-Gal) in the digestive tract, humans are unable to digest these sugars, which consequently induce flatulence. To develop the consumption of the otherwise highly nutritional soy products, the use of exogenous alpha-Gal is promising. In this framework, we characterized the melA gene for alpha-Gal in Lactobacillus plantarum. The melA gene encodes a cytoplasmic 84-kDa protein whose enzymatically active form occurs as oligomers. The melA gene was cloned and expressed in Escherichia coli, yielding an active alpha-Gal. We show that melA is transcribed from its own promoter, yielding a monocistronic mRNA, and that it is regulated at the transcriptional level, i.e., it is induced by melibiose but is not totally repressed by glucose. Post-transcriptional regulation by the carbon source could also occur. Upstream of melA, a putative galactoside transporter, designated RafP, was identified that shows high homology to LacS, the unique transporter for both alpha- and alpha-galactosides in Streptococcus thermophilus. rafP is also expressed as a monocistronic mRNA. Downstream of melA, the lacL and lacM genes were identified that encode a heterodimeric alpha-galactosidase. A putative galM gene identified in the same cluster suggests the presence of a galactose operon. These results indicate that the genes involved in galactoside catabolism are clustered in L. plantarum ATCC 8014. This first genetic characterization of melA and of its putative associated transporter, rafP, in a lactobacillus opens doors to various applications both in the manufacture of soy-derived products and in probiotic and nutraceutical issues. Fil: Silvestroni, Aurelio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Connes, Cristelle. Institut National de la Recherche Agronomique; Francia Fil: Sesma, Fernando Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina Fil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Microbiología; Argentina Fil: Piard, Jean-Christophe. Institut National de la Recherche Agronomique; Francia