Acid tolerance mediated by membrane ATPases in Lactobacillus acidophilus

The acid tolerance response in Lactobacillus acidophilus CRL 639, induced at pH 4.2 for 15 min, is mediated by the cell membrane F1-F0 ATPase. The specific activity of the enzyme was induced 1.6-fold after acid adaptation compared to a non-adapted control. The ATPase was optimal at pH 6 with a Km = 0.8 mM and a Vmax = 100 mM.Fil: Lorca, Graciela L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Novel pathway for corrinoid compounds production in lactobacillus

Vitamin B12 or cobalamin is an essential metabolite for humans, which makes it an interesting compound for many research groups that focus in different producer-strains synthesis pathways. In this work, we report the influence of key intermediaries for cobalamin synthesis added to the culture medium in two Lactobacillus (L.) strains, L. reuteri CRL 1098 and L. coryniformis CRL 1001. Here, we report that addition of Co2+ and 5,6-dimethylbenzimidazole increased the corrinoid compounds production in both strains while addition of L-threonine increased only the corrinoid compounds production by CRL 1001 strain. Then, we purified and characterized by LC-MS the corrinoid compounds obtained. Physiological studies besides in silico analysis revealed that L. reuteri CRL 1098 and L. coryniformis CRL 1001 follow different pathways for the last steps of the corrinoid compounds synthesis.Fil: Torres, Andrea Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vannini, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Font, Graciela Maria. 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; ArgentinaFil: Saavedra, Maria Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Taranto, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Biopolymers from lactic acid bacteria. Novel applications in foods and beverages

Lactic acid bacteria (LAB) are microorganisms widely used in the fermented food industry worldwide. Certain LAB are able to produce exopolysaccharides (EPS) either attached to the cell wall (capsular EPS) or released to the extracellular environment (EPS). According to their composition, LAB may synthesize heteropolysaccharides or homopolysaccharides. A wide diversity of EPS are produced by LAB concerning their monomer composition, molecular mass, and structure. Although EPS-producing LAB strains have been traditionally applied in the manufacture of dairy products such as fermented milks and yogurts, their use in the elaboration of low-fat cheeses, diverse type of sourdough breads, and certain beverages are some of the novel applications of these polymers. This work aims to collect the most relevant issues of the former reviews concerning the monomer composition, structure, and yields and biosynthetic enzymes of EPS from LAB; to describe the recently characterized EPS and to present the application of both EPS-producing strains and their polymers in the fermented (specifically beverages and cereal-based) food industry.Fil: Torino, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Mozzi, Fernanda Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Genome sequence of the cheese-starter strain Lactobacillus delbrueckii subsp. lactis CRL 581

We report the genome sequence of Lactobacillus delbrueckii subsp. lactis CRL 581 (1,911,137 bp, GC 49.7%), a proteolytic strain isolated from a homemade Argentinian hard cheese which has a key role in bacterial nutrition and releases bioactive healthbeneficial peptides from milk proteins.Fil: Hebert, Elvira Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Raya, Raul Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Brown, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Taranto, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina

Influencia del aceite esencial de orégano en la elaboración tradicional de quesos: Efecto sobre el fermento láctico

The aim of this work is to study the oregano essential oil (OEO) composition from Northwestern Argentinean regions and to evaluate its effect on the lactic starter cultures. The oregano used, Origanum vulgare var hirtum, was obtained from Andalgalá, Catamarca. The essential oil presented high amounts of α-terpinene (10%), γ-terpinene (15.1%), terpinen- 4-ol (15.5%) and thymol (13.0%) as the main components. No negative effect on growth or metabolic activity of lactic acid bacteria Streptococcus thermophilus CRL 728 and CRL 813, Lactobacillus delbrueckii subsp. bulgaricus CRL 656 and CRL 468, and Lactococcus lactis subsp. lactis CRL 597 up to the maximum concentration (200 μg/g) assayed was observed. No differ-ences in the organoleptic characteristics of semi-hard cheeses flavored with oregano essential oil (200 μg/g) and homemade cheeses flavored with oregano leaves were found. With respect to the microbiological quality of the products, neither enterobacteria nor mold and yeast were detected during ripening in essential-oil flavored cheese compared to control cheese (enter-obacteria 2 × 103 UFC/g) and cheese flavored with oregano leaves (mold/yeast 4 × 104 CFU/g). Our results showed that the use of oregano essential oil and lactic starter culture considerably improved cheese quality.Fil: Marcial, Guillermo Emilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Gerez, Carla Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Nuñez, Martha Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Coll Araoz, Maria Victoria. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Schuff, Carola. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Font, Graciela Maria. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Draft genome sequence of Lactobacillus rhamnosus CRL1505, an immunobiotic strain used in social food programs in Argentina

We report the draft genome sequence of the probiotic Lactobacillus rhamnosus strain CRL1505. This new probiotic strain has been included into official Nutritional Programs in Argentina. The draft genome sequence is composed of 3,417,633 bp with 3,327 coding sequences.Fil: Taranto, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina;Fil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Universidad Nacional de Tucuman. Facultad de Bioquimica, Quimica y Farmacia. Instituto de Bioquimica Clinica Aplicada; Argentina;Fil: Savoy, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina; Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina;Fil: Hebert, Elvira Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina

Genomic analysis of vitamin B12 (Cobalamin) biosynthetic cluster in Lactobacillus coryniformis CRL 1001

The vitamin B12 (cobalamin-CBL), a very complex non-polymeric macromolecule synthesized only by some bacteria and archaea, and essential to humans and animals. The highly complicated CBL biosynthesis involves about 30 synthesis steps. We demonstrated that cell extract of Lactobacillus coryniformis CRL 1001 is able to correct the coenzyme B12 requirement of Salmonella enterica serovarTyphimuriumin minimal medium. The aim of this study was the sequencing of CRL1001 genome and the molecular characterization of CBL biosynthesis in this strain. L. coryniformis CRL 1001 genome was sequenced by a whole-genome shotgun (WGS) strategy with an Ion Torrent personal genome machine based upon libraries created using NEBNext DNA library kits. Genomic analysis was done using the RAST annotation Server, Blast algorithms, ISGA and KEGG databases.The draft genome sequence consists of 2,829,178 bases with a mean GC content of 42%. A total of 3,341 coding sequences (CDS) and 82 structural RNAs (58 tRNAs) were predicted. RAST analysis evidenced the presence of at least 30 genes (cob genes) involved in the CBL biosynthesis in CRL 1001 strain. This finding is the first evidence for cobalamin biosynthesis genes in this species. Comparative studies among vitamin B12 producer strains demonstrated that the genetic organization of cob operon is conserved in this strain and these genes are adjacent to the pdu operon and pocR gene. The hem genes (hem A, C, B y L) present in L. coryniformis CRL 1001 genome are located among cob operon in similar way to anaerobic microorganisms. Interestingly, the cblT y cblS genes were identified in CRL1001 strain genome. These genes encode a putative protein kinase and a α-ribasol transporter, respectively. The cblT y cblS genes are present in CBLproducers Listeria sp. strains but absent in CBL-producers L. reuteri strains. The knowledge of cob genes and their regulation in vitamin producer lactic strains constitutes an interesting biotechnological alternative for developing fortified foods.Fil: Torres, Andrea Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vannini, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Sesma, Fernando Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Saavedra, Maria Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Taranto, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaX Congreso Argentino de Microbiología GeneralMar del PlataArgentinaSociedad Argentina de Microbiología Genera

Lactic acid bacteria from artisanal tanneries: isolation and evaluation of acidifyng ability

The tanning process allows to transform animal skin in stable and non-putrescible products namely leather. This process consists in various steps such as washing skins, liming, depilation, lime removal by washing, purging and tanning. During purging, the skins are immersed in a cereal mix which is let to ferment for 12-24 h at room temperature (18ºC-37°C). After this period, the pH decreased to 4.5-5.0, a condition that is required for the final steps. Under these homemade working conditions, the cereal fermentation is quite variable and affects the quality of the leather. The objective of this study was to isolate lactic acid bacteria (LAB) from the fermented cereal mixture in artisanal tanneries and to evaluate in vitro the acidifying activity of the strains. This is the first step for the formulation of a starter culture for tanneries to normalize the process.Samples of fermented cereals were taken from a tannery located in San Pedro de Colalao, Tucumán, Argentina, and colonies were isolated in MRS agar medium. The primary identification of the isolates included Gram stain, microscopic observations and catalase reaction. Gram positive, catalase negative strains (23) were cultured in a CERELA medium formulated for the production of lactic ferment and incubated at 18°C and 37°C for 24 h. At intervals, pH and titratable acidity (TTA) were determined. In most samples, a prevalence of cocci (95%) respect to bacilli was observed. The total isolated strains (56), 23 strains were Gram (+) and catalase (negative) which were selected. At 37 °C, most strains (21) acidified the culture medium within the first 8-h reaching a final pH ≤ 5.0 and a TTA ≥100ºD. At 18 °C, all strains showed a lower growth; however, the decrease in pH was 4.5-5.0 and the acidity developed (100-140ºD) after 24 hours of fermentation were similar to values obtained at 37°C. The best acid-producer strains were identified as Enterococcus faecium CRL 1943 and Leuconostoc citreum CRL 1945 by phenotypic and genotypic techniques. Currently, studies are being conducted to formulate a lactic inoculant on the basis of these strains and their metabolites for artisanal tanning.Fil: Correa Deza, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Gerez, Carla Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Martos, Gladys Irma. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Nuñez, Martha Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Fiori, Mario. Instituto Nacional de Tecnología Industrial; ArgentinaFil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaXI Congreso Argentino de Microbiología GeneralCórdobaArgentinaSociedad Argentina de Microbiología Genera

Physicochemical composition and microbiological quality of goat milk produced in Santiago del Estero Province (Argentine)

La mayor cuenca lechera caprina de Argentina está ubicada en la provincia de Santiago del Estero, pero existe escaso conocimiento de la calidad de leche producida allí. El presente trabajo constó de dos fases y tuvo como objetivo analizar la composición fisicoquímica y calidad microbiológica de la leche de cabra de esta cuenca lechera. En la primera fase se analizó la leche procedente de 30 pequeños tambos sin raza animal definida y se obtuvieron los siguientes valores medios: grasa, 5.37%; proteína, 3.39%; lactosa, 4.32%; sólidos no grasos (SNG), 8.30%; sólidos totales (ST), 13.66%; acidez, 17.61ºD; pH, 6.76; densidad, 1.031g/mL; aerobios mesófilos totales, 4.96 log ufc/mL y proporción de muestras positivas de coliformes fecales, 72%. En la segunda fase, se seleccionaron tres tambos con razas lecheras definidas y volúmenes de producción superiores a la media de la zona. La leche de estos tambos no difirió mucho de la composición fisicoquímica ya citada, pero se destacaron dos de los mismos (T2 y T3) por sus respectivos contenidos de grasa (5.59 y 5.52%) y ST (14.18 y 13.84%), resultado de su uso de la raza Anglo Nubian y alimentación mediante pastoreo de pasturas naturales y suplementación con cereales. El T1, que usa la raza Saanen presentó la mayor producción media de leche, de 1.44 kg/d y la acidez más baja, de 16.49ºD, resultado de mejores condiciones de manejo del tambo y enfriamiento de la leche postordeño. Se concluye que aun haciendo falta mejorar el manejo higiénicosanitario que recibe la leche cruda, la leche de esta cuenca se presta muy bien para la elaboración de queso.The largest caprine milk shed of Argentina is located in Santiago del Estero province, but little information is available about the quality of the milk produced there. The present study consisted of two phases and had the objective of analyzing the physical-chemical composition and microbiological quality of the goat’s milk of this milk shed. In the first phase, milk from 30 small producer operations without defined animal breeds were analyzed with the following mean results: fat, 5.37%; protein, 3.39%; lactose, 4.32%; solids/not/fat (SNF), 8.30%; total solids (TS) 13.66%; acid degree value, 17.61ºD; pH, 6.76; density, 1.031 g/mL; total aerobic mesophiles, 4.96 log cfu/mL and proportion of samples positive for fecal coliforms, 72%. The second phase of the study involved three goat farms with different breeds and production levels exceeding the average of the zone. The milk from these farms did not differ much from the physical-chemical composition cited above, but two of them (T2 and T3) had outstanding contents of fat (5.59 and 5.52%) and TS (14.13 and 13.84%), resulting from their use of the Anglo Nubian breed and feeding based on grazing of natural pastures and supplementation with cereals. The farm T1, that employs the Saanen breed, had the highest mean milk production (1.44 kg/day) and the lowest acid degree value (16.49ºD), resulting from better management of the operation and more efficient cooling of the milk after milking. It is concluded that, eventhough better hygienic/sanitary management of the crude milk is needed, milk from this milk shed is well suited for cheese making.Fil: Frau, Silvia Florencia. Universidad Nacional de Santiago del Estero; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Font, Graciela Maria. Universidad Nacional de Santiago del Estero; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Paz, Raul Gustavo. Universidad Nacional de Santiago del Estero; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pece Azar, Nora Beatriz del Carmen. Universidad Nacional de Santiago del Estero; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Inorganic polyphosphate from the immunobiotic Lactobacillus rhamnosus CRL1505 prevents inflammatory response in the respiratory tract

Lactobacillus (L.) rhamnosus CRL1505 accumulates inorganic polyphosphate (polyP) in its cytoplasm in response to environmental stress. The aim of this study was to evaluate the potential effects of polyP from the immunobiotic CRL1505 on an acute respiratory inflammation murine animal model induced by lipopolysaccharide (LPS). First, the presence of polyP granules in the cytoplasm of CRL1505 strain was evidenced by specific staining. Then, it was demonstrated in the intracellular extracts (ICE) of CRL1505 that polyP chain length is greater than 45 phosphate residues. In addition, the functionality of the genes involved in the polyP metabolism (ppk, ppx1 and ppx2) was corroborated by RT-PCR. Finally, the possible effect of the ICE of CRL1505 strain containing polyP and a synthetic polyP was evaluated in vivo using a murine model of acute lung inflammation. It was observed that the level of cytokines pro-inflammatory (IL-17, IL-6, IL-2, IL-4, INF-γ) in serum was normalized in mice treated with ICE, which would indicate that polyP prevents the local inflammatory response in the respiratory tract. The potential application of ICE from L. rhamnosus CRL1505 as a novel bioproduct for the treatment of respiratory diseases is one of the projections of this work.Fil: Correa Deza, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Rodríguez de Olmos, Antonieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Suárez, Nadia Elina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Gerez, Carla Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin