Characterization of autochthonous lactobacilli from goat dairy products with probiotic potential for metabolic diseases.

The present study aimed to design functional fermented goat milk with probiotic potential for metabolic diseases. Thereby, autochthonous lactobacilli from goat dairy products that target improving the inflammatory, lipid, and glycemic profile were characterized. We designed fermented goat milk using Lactobacillus delbrueckii subsp. indicus CRL1447 as starter strain, supplemented with different probiotic consortia formed by Limosilactobacillus fermentum CRL1446, Lactiplantibacillus paraplantarum CRL1449, and CRL1472 strains. These lactobacilli were selected for their positive effects on inhibition of α-glucosidase, bile salts hydrolase activity, cholesterol assimilation, and decreased triglyceride percentage in Caenorhabditis elegans. Furthermore, the lactobacilli oral administration to obese mice caused a significant decrease in body weight gain and ameliorated hyperglycemia and hyperlipemia. These results reveal the potential of this goat dairy product as a functional food to prevent obesity and related pathologies. Goat milk-derived products stand out for their marketing potential. Hence, fermented goat milk incorporating novel probiotics represents a group of food products with broad prospects by their promising nutritive and therapeutic properties for metabolic diseases. The goat dairy product designed in this study could be used in the prevention of dyslipidemia and hyperglycemia in obese people.

Ferulic Acid Esterase Producing Lactobacillus johnsonii from Goat Feces as Corn Silage Inoculants.

Ferulic acid esterase (FAE+)-producing lactobacilli are being studied as silage inoculants due to their potential of increasing forage fiber digestibility. In this work, three FAE+ Lactobacillus (L.) johnsonii strains were isolated from caprine feces and characterized according to their potential probiotic characteristics and as silage inoculants. Limosilactobacillus fermentum CRL1446, a human probiotic isolated from goat cheese, was also included in the experiments as a potential silage inoculant. FAE activity quantification, probiotic characterization, and growth in maize aqueous extract indicated that L. johnsonii ETC187 might have a better inoculant and probiotic aptitude. Nevertheless, results in whole-corn mini silos indicated that, although acid detergent fiber (ADF) was significantly reduced by this strain (3% compared with the uninoculated (UN) group), L. johnsonii ETC150 and CRL1446 not only induced similar ADF reduction but also reduced dry matter (DM) loss (by 7.3% and 6.5%, respectively) compared with the UN group. Additionally, CRL1446 increased in vitro DM degradability by 10%. All treatments reduced gas losses when compared with the UN group. The potential probiotic features of these strains, as well as their beneficial impact on corn fermentation shown in this study, encourage further studies as enhancers in animal production.

Aroma compounds and consumer acceptability of soybean paste fermented by lactobacilli.

Consumption of soybean-based foods is affected by the flavor of the legume; due to the presence of undesirable compounds called "beany flavors". To solve this problem, the influence of solid state fermentation by lactobacilli on the production of volatile compounds in soybean paste was determined. The volatile's production was measured by gas chromatography. Forty compounds were identified in the different soybean pastes studied. The results showed that fermentation stimulates the production of desirable volatile compounds in foods such as ketones (22-75%) and decreased unpleasant compounds (10-84%).The consumers acceptance study showed that a group of participants (30% approximately) preferred the fermented samples associated with sweet and acid aroma like yogurt. In conclusion, the fermentation positively influences the generation of desirable volatile compounds and completely reduces hexanal in one sample. Therefore, fermentation with the studied strains is a valid strategy to modify the aroma profile of a soybean-based food matrix. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05210-5.

Ability of Boron to Act as a Nucleophile and an Electrophile in Boryl Shift Reactions Unveiled by Electron Density Distribution Analysis.

The behavior of the tetracoordinate boron of N-methyliminodiacetic acid (MIDA) boronates as a nucleophile and an electrophile during the 1,2-boryl migration promoted by a Lewis acid and the 1,4-boryl migration promoted by a neighboring atom, respectively, have been investigated using density functional theory and the quantum theory of atoms in molecules. We found that when boron acts as a nucleophile, the electron density of the B-N interaction of the BMIDA moiety maintains the charge concentration over the boron atom, facilitating its transport toward the electron-deficient center. In this process, the BMIDA remains as a tetracoordinate. On the other hand, the B-N weakening generates a charge depletion region over the boron, allowing it to interact with the electron-rich center of O1, developing the boron atom, a pentacoordinate form. Then, the B-N bond breaking triggers a series of changes in the electronic structure of the boron atom. Our results explain the role of the MIDA ligand upon the remarkable susceptibility of the boron atom for switching its structural and electronic characteristics in the migration processes. In addition, the dichotomous behavior was evaluated with a different scenario, considering tricoordinate pinacol boronate as a boryl migrating group.

Spray dried lactobacilli maintain viability and feruloyl esterase activity during prolonged storage and under gastrointestinal tract conditions.

The use of lactobacilli with feruloyl esterase (FE) activity in the development of functional foods has gained considerable interest in recent years. Microencapsulation of FE-producing bacteria to facilitate their incorporation into food is a challenge. The aim of this study was to evaluate survival and maintenance of FE activity during storage at 4 °C and under simulated gastrointestinal tract (GIT) conditions of microcapsules of FE-producing Lactobacillus (Lb.) strains obtained by spray drying. Lb. fermentum CRL1446 and Lb. johnsonii CRL1231 powders maintained viability at concentrations ≥ 106 CFU/g (minimum probiotic dose) when stored at 4 °C for 12 months. Lb. acidophilus CRL1014 powders were only able to maintain ≥ 106 CFU/g during 4 months of storage. FE activity was conserved in three microencapsulated strains evaluated, an increase of specific activity being observed until month 12 of storage. Powders of the three strains incubated under GIT conditions maintained their viability (≥ 106 CFU/g), but specific FE activity was only detected in Lb. fermentum and Lb. johnsonii powders (0.80-0.83 and 0.21-0.56 U/mg, respectively). CRL1446 and CRL1231 microcapsules were able to resist prolonged storage and GIT conditions, retaining FE activity and preserving their probiotic potential and could be incorporated into functional foods.

Lactic Acid Bacteria Strains Differently Modulate Gut Microbiota and Metabolic and Immunological Parameters in High-Fat Diet-Fed Mice.

Background: Dietary strategies, including the use of probiotics as preventive agents that modulate the gut microbiota and regulate the function of adipose tissue, are suitable tools for the prevention or amelioration of obesity and its comorbidities. We aimed to evaluate the effect of lactic acid bacteria (LAB) with different adipo- and immuno-modulatory capacities on metabolic and immunological parameters and intestinal composition microbiota in high-fat-diet-induced in mice fed a high-fat diet Methods: Balb/c weaning male mice were fed a standard (SD) or high-fat diet (HFD) with or without supplementation with Limosilactobacillus fermentum CRL1446 (CRL1446), Lactococcus lactis CRL1434 (CRL1434), or Lacticaseibacillus casei CRL431 (CRL431) for 45 days. Biochemical and immunological parameters, white-adipose tissue histology, gut microbiota composition, and ex vivo cellular functionality (adipocytes and macrophages) were evaluated in SD and HFD mice. Results: CRL1446 and CRL1434 administration, unlike CRL431, induced significant changes in the body and adipose tissue weights and the size of adipocytes. Also, these strains caused a decrease in plasmatic glucose, cholesterol, triglycerides, leptin, TNF-α, IL-6 levels, and an increase of IL-10. The CRL1446 and CRL1434 obese adipocyte in ex vivo functionality assays showed, after LPS stimulus, a reduction in leptin secretion compared to obese control, while with CRL431, no change was observed. In macrophages from obese mice fed with CRL1446 and CRL1434, after LPS stimulus, lower levels of MCP-1, TNF-α, IL-6 compared to obese control were observed. In contrast, CRL431 did not induce modification of cytokine values. Regarding gut microbiota, all strain administration caused a decrease in Firmicutes/Bacteroidetes index and diversity. As well as, related to genus results, all strains increased, mainly the genera Alistipes, Dorea, Barnesiella, and Clostridium XIVa. CRL1446 induced a higher increase in the Lactobacillus genus during the study period. Conclusions: The tested probiotic strains differentially modulated the intestinal microbiota and metabolic/immunological parameters in high-fat-diet-induced obese mice. These results suggest that CRL1446 and CRL1434 strains could be used as adjuvant probiotics strains for nutritional treatment to obesity and overweight. At the same time, the CRL431 strain could be more beneficial in pathologies that require regulation of the immune system.

Efecto antibacteriano de una pasta dental con xilitol sobre Streptococcus mutans en saliva de gestantes / Antibacterial effect of a toothpaste with xylitol on Streptococcus mutans in pregnant women's saliva

RESUMEN Introducción: Durante la gestación, algunos cambios fisiológicos en la mujer propician un incremento de la placa dental y de la microbiota cariogénica. Esta condición unida a una modificación de la dieta y a una inadecuada higiene bucal condicionarían desfavorablemente la salud oral tanto de la madre como del bebé. Objetivo: Determinar el efecto antibacteriano de una pasta dental con xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes. Métodos: Ensayo clínico, a doble ciego, que se realizó en el Centro de Salud "José Olaya" (Chiclayo Perú), en enero de 2017. Se trabajó con una población muestral de 50 gestantes en el segundo trimestre que cumplieron con los criterios establecidos. Se crearon dos grupos: 25 gestantes usaron dentífrico con 10 por ciento de xilitol y 25 gestantes usaron dentífrico sin xilitol. Se les tomó y proceso microbiológicamente una muestra de saliva antes del inicio del estudio y 14 días después del uso de las respectivas pastas. Se realizó el recuento de unidades formadoras de colonias de Streptococcus mutans en saliva con una confiabilidad altamente significativa mediante el coeficiente de correlación intraclase, calibración intra e interexaminador (1,000 y 0,999, respectivamente). El análisis de los datos se realizó mediante la prueba U de Mann-Whitney, considerando un nivel de significancia del 5 por ciento. Resultados: No se encontró diferencia significativa entre las gestantes que emplearon pasta dental con xilitol en comparación con las que utilizaron pasta dental sin xilitol (p= 0,062). Conclusiones: El efecto antibacteriano de la pasta dental comercial con xilitol es similar a una pasta dental sin xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes(AU)

ABSTRACT Introduction: Some physiological changes occur in women during pregnancy which cause an increase in dental plaque and cariogenic microbiota. This situation, alongside a modification in the diet and inadequate oral hygiene, negatively affect the oral health of both the mother and the baby. Objective: Determine the antibacterial effect of a toothpaste with xylitol on the count of Streptococcus mutans in pregnant women's saliva. Methods: A double-blind clinical trial was conducted in José Olaya Health Center (Chiclayo, Peru) in January 2017. The sample population was 50 women in the second trimester of pregnancy who met the established inclusion criteria. Two groups were formed: 25 pregnant women used a toothpaste with 10 percent xylitol and the other 25 used a toothpaste without xylitol. A saliva sample was taken and processed microbiologically before the start of the study and 14 days after use of the two toothpastes. A count was performed of colony-forming units for Streptococcus mutans in saliva with highly significant reliability, using the intraclass correlation coefficient, and intra- and inter-rater calibration (1.000 and 0.999, respectively). Data analysis was based on the Mann-Whitney U Test, with a significance level of 5 percent. Results: No significant difference was found between the pregnant women who used the toothpaste with xylitol and those who used the toothpaste without xylitol (p= 0.062). Conclusions: The antibacterial effect of the commercial toothpaste with xylitol is similar to that of a toothpaste without xylitol in terms of the Streptococcus mutans count in pregnant women's saliva(AU)

Efecto de una Pasta Dental Comercial Conteniendo Xilitol Sobre el Recuento de Streptococcus Mutans en Saliva de Gestantes: Ensayo Clínico Controlado Aleatorizado / Effect of a Commercial Toothpaste Containing Xylitol on the Count of Streptococcus Mutans in Saliva of Pregnant Women: Randomized Controlled Clinical Trial

RESUMEN: El objetivo del presente trabajo fue determinar el efecto de una pasta dental comercial conteniendo xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes. El presente fue un ensayo clínico, a doble ciego, que se realizó en el Centro de Salud "José Olaya" (Chiclayo Perú), en enero de 2017. Se trabajó con una población muestral de 50 gestantes en el segundo trimestre que cumplieron con los criterios establecidos, distribuyéndolas en dos grupos: 25 gestantes usaron pasta dental con 10 % de xilitol y 25 gestantes usaron pasta dental sin xilitol. Se les tomó y procesó microbiológicamente una muestra de saliva antes del inicio del estudio y 14 días después del uso de las respectivas pastas. Se realizó el recuento de unidades formadoras de colonias (UFC) de Streptococcus mutans en saliva con una confiabilidad altamente significativa mediante el Coeficiente de Correlación Intraclase, calibración intra e interexaminador (1,000 y 0,999, respectivamente). El análisis de los datos se realizó mediante la prueba U de Mann-Whitney, considerando un nivel de significancia del 5 %. No se encontró diferencia entre las gestantes que emplearon pasta dental con xilitol en comparación con las que utilizaron pasta sin xilitol (p=0,062). Se concluyó que el efecto de la pasta dental comercial conteniendo xilitol es similar a una pasta sin xilitol sobre el recuento de Streptococcus mutans en saliva de gestantes.

ABSTRACT: The objective of the present study was to determine the effect of a commercial toothpaste containing xylitol on the counts of Streptococcus mutans in saliva of pregnant women. The present was a double-blind clinical trial performed at the "José Olaya" health Centre (Chiclayo Peru) in January 2017. We worked with a sample population of 50 pregnant in the second trimester that met the established criteria, distributing in two groups: 25 pregnant women used toothpaste with 10 % xylitol and 25 pregnant used toothpaste without xylitol. They were taken and microbiologically processed a sample of saliva before the start of the study and 14 days after the use of the respective toothpastes. The Colony-forming units (CFU) of Streptococcus mutans in saliva were counted with a highly significant reliability through the intraclass correlation coefficient, Intra-and Interexaminer calibration (1.000 and 0.999, respectively). Data analysis was performed using the Mann-Whitney U test, considering a 5 % significance level. No difference was found among the pregnant women who used xylitol toothpaste compared to those who used toothpaste without xylitol (p = 0,062). It was concluded that the effect of xylitol containing commercial toothpaste is similar to a toothpaste without xylitol on the count of Streptococcus mutans in the saliva of pregnant women.

Diversity and Functional Properties of Lactic Acid Bacteria Isolated From Wild Fruits and Flowers Present in Northern Argentina.

Lactic acid bacteria (LAB) are capable of converting carbohydrate substrates into organic acids (mainly lactic acid) and producing a wide range of metabolites. Due to their interesting beneficial properties, LAB are widely used as starter cultures, as probiotics, and as microbial cell factories. Exploring LAB present in unknown niches may lead to the isolation of unique species or strains with relevant technological properties. Autochthonous rather than allochthonous starter cultures are preferred in the current industry of fermented food products, due to better adaptation and performance of autochthonous strains to the matrix they originate from. In this work, the lactic microbiota of eight different wild tropical types of fruits and four types of flowers were studied. The ability of the isolated strains to produce metabolites of interest to the food industry was evaluated. The presence of 21 species belonging to the genera Enterococcus, Fructobacillus, Lactobacillus, Lactococcus, Leuconostoc, and Weissella was evidenced by using culture-dependent techniques. The isolated LAB corresponded to 95 genotypically differentiated strains by applying rep-PCR and sequencing of the 16S rRNA gene; subsequently, representative strains of the different isolated species were studied for technological properties, such as fast growth rate and acidifying capacity; pectinolytic and cinnamoyl esterase activities, and absence of biogenic amine biosynthesis. Additionally, the strains' capacity to produce ethyl esters as well as mannitol was evaluated. The isolated fruit- and flower-origin LAB displayed functional properties that validate their potential use in the manufacture of fermented fruit-based products setting the background for the design of novel functional foods.

Draft Genome Sequence of the Feruloyl Esterase-Producing Strain Lactobacillus fermentum CRL1446, a Probiotic for Malnutrition.

We report here the draft genome sequence of Lactobacillus fermentum CRL1446 (2,148,781 bp, 51.4% G+C content). This strain exhibits feruloyl esterase activity and important technological and probiotic properties. Because of its proven beneficial effects in vivo, it represents an interesting candidate for the development of functional foods or pharmabiotics for malnutrition.

Specific Strains of Lactic Acid Bacteria Differentially Modulate the Profile of Adipokines In Vitro.

Obesity induces local/systemic inflammation accompanied by increases in macrophage infiltration into adipose tissue and production of inflammatory cytokines, chemokines, and hormones. Previous studies have shown that probiotics could improve the intestinal dysbiosis induced by metabolic diseases such as obesity, diabetes, and metabolic syndrome. Microorganisms could (directly or indirectly) affect adipokine levels due to their capacity to induce translocation of several intestinal microbial antigens into systemic circulation, which could lead to metabolic endotoxemia or produce immunomodulation in different organs. The aim of the present study was to select non-inflammatory lactic acid bacteria (LAB) strains with the capacity to modulate adipokine secretion by the adipose tissue. We wish to elucidate the role of potential probiotic strains in the regulation of the cross talking between immune cells such as macrophages and adipose cells. Mouse macrophage cell line RAW 264.7 was used for evaluating the ability of 14 LAB strains to induce cytokine production. The LAB strains were chosen based on their previously studied beneficial properties in health. Then, in murine adipocyte culture and macrophage-adipocyte coculture, we determined the ability of these strains to induce cytokines and leptin secretion. Tumor necrosis factor alpha, interleukin 6 (IL-6), IL-10, monocyte chemoattractant protein-1, and leptin levels were measured in cell supernatants. We also performed the detection and quantification of leptin receptor (Ob-Rb) expression in macrophage cell lines stimulated by these LAB strains. Differential secretion profile of cytokines in macrophage cells induced by LAB strains was observed. Also, the levels of Ob-Rb expression diverged among different LAB strains. In LAB-stimulated coculture cells (adipocytes and macrophages), we observed differential production of leptin and cytokines. Furthermore, we detected lower production levels in single culture than cocultured cells. The principal component analysis showed an association between the four clusters of strains established according to their inflammatory profiles and leptin adipocyte production and leptin receptor expression in macrophages. We conclude that coculture is the most appropriate system for selecting strains with the ability to modulate adipokine secretion. The use of microorganisms with low and medium inflammatory properties and ability to modulate leptin levels could be a strategy for the treatment of some metabolic diseases associated with dysregulation of immune response.

Lactobacillus fermentum CRL1446 Ameliorates Oxidative and Metabolic Parameters by Increasing Intestinal Feruloyl Esterase Activity and Modulating Microbiota in Caloric-Restricted Mice.

The purpose of this study was to determine whether the administration of the feruloyl esterase (FE)-producing strain Lactobacillus fermentum CRL1446 enhances metabolic and oxidative parameters in caloric-restricted (CR) mice. Balb/c male mice were divided into ad libitum fed Group (ALF Group), CR diet Group (CR Group) and CR diet plus L. fermentum Group (CR-Lf Group). CR diet was administered during 45 days and CRL1446 strain was given in the dose of 108 cells/mL/day/mouse. FE activity was determined in intestinal mucosa and content at Day 1, 20 and 45. Triglyceride, total cholesterol, glucose, thiobarbituric acid reactive substances (TBARS) levels and glutathione reductase activity were determined in plasma. Gut microbiota was evaluated by high-throughput sequencing of 16S rRNA gene amplicons. At Day 45, total intestinal FE activity in CR-Lf Group was higher (p = 0.020) than in CR and ALF groups and an improvement in both metabolic (reductions in triglyceride (p = 0.0025), total cholesterol (p = 0.005) and glucose (p < 0.0001) levels) and oxidative (decrease of TBARS levels and increase of plasmatic glutathione reductase activity (p = 0.006)) parameters was observed, compared to ALF Group. CR diet increased abundance of Bacteroidetes and CRL1446 administration increased abundance of Bifidobacterium and Lactobacillus genus. L. fermentun CRL1446 exerted a bifidogenic effect under CR conditions.

Novel rat model for neurocysticercosis using Taenia solium.

Neurocysticercosis is caused by Taenia solium infecting the central nervous system and is the leading cause of acquired epilepsy and convulsive conditions worldwide. Research into the pathophysiology of the disease and appropriate treatment is hindered by lack of cost-effective and physiologically similar animal models. We generated a novel rat neurocysticercosis model using intracranial infection with activated T. solium oncospheres. Holtzman rats were infected in two separate groups: the first group was inoculated extraparenchymally and the second intraparenchymally, with different doses of activated oncospheres. The groups were evaluated at three different ages. Histologic examination of the tissue surrounding T. solium cysticerci was performed. Results indicate that generally infected rats developed cysticerci in the brain tissue after 4 months, and the cysticerci were observed in the parenchymal, ventricle, or submeningeal brain tissue. The route of infection did not have a statistically significant effect on the proportion of rats that developed cysticerci, and there was no dependence on infection dose. However, rat age was crucial to the success of the infection. Epilepsy was observed in 9% of rats with neurocysticercosis. In histologic examination, a layer of collagen tissue, inflammatory infiltrate cells, perivascular infiltrate, angiogenesis, spongy change, and mass effect were observed in the tissue surrounding the cysts. This study presents a suitable animal model for the study of human neurocysticercosis.

The secreted esterase of Propionibacterium freudenreichii has a major role in cheese lipolysis.

Free fatty acids are important flavor compounds in cheese. Propionibacterium freudenreichii is the main agent of their release through lipolysis in Swiss cheese. Our aim was to identify the esterase(s) involved in lipolysis by P. freudenreichii. We targeted two previously identified esterases: one secreted esterase, PF#279, and one putative cell wall-anchored esterase, PF#774. To evaluate their role in lipolysis, we constructed overexpression and knockout mutants of P. freudenreichii CIRM-BIA1(T) for each corresponding gene. The sequences of both genes were also compared in 21 wild-type strains. All strains were assessed for their lipolytic activity on milk fat. The lipolytic activity observed matched data previously reported in cheese, thus validating the relevance of the method used. The mutants overexpressing PF#279 or PF#774 released four times more fatty acids than the wild-type strain, demonstrating that both enzymes are lipolytic esterases. However, inactivation of the pf279 gene induced a 75% reduction in the lipolytic activity compared to that of the wild-type strain, whereas inactivation of the pf774 gene did not modify the phenotype. Two of the 21 wild-type strains tested did not display any detectable lipolytic activity. Interestingly, these two strains exhibited the same single-nucleotide deletion at the beginning of the pf279 gene sequence, leading to a premature stop codon, whereas they harbored a pf774 gene highly similar to that of the other strains. Taken together, these results clearly demonstrate that PF#279 is the main lipolytic esterase in P. freudenreichii and a key agent of Swiss cheese lipolysis.

Contribution of lactic acid bacteria esterases to the release of fatty acids in miniature ewe's milk cheese models.

The present work evaluates the contribution of esterase activities of lactic acid bacteria isolated from ewe's dairy products to the release of free fatty acids (FFA) in ewe's milk cheese models. At 60 days of ripening, single-strain cheeses Ov 409 and Ov 421 showed high levels of total FFA (3075 and 2494.62 mg/kg, respectively). Cheeses Ov 227-Ov 409 and Ov 421-Ov 409 presented high percentages of short-chain fatty acids (SCFA). The highest levels of volatile free fatty acids (VFFA) were detected in cheeses Ov 409, Ov 421-Ov 409, and Ov 421-Ov 227. Studies on esterase activities showed that these strains hydrolyzed alpha-naphthyl derivatives of fatty acids from C2 to C6, mainly associated with the wall-membrane fraction. The results showed that the strains studied contributed to the release of FFA during ripening of ewe's milk cheese models. The increase of SCFA throughout ripening involves the action of esterases of starter strains.

Technological properties of Enterococcus faecium isolated from ewe's milk and cheese with importance for flavour development.

Eight Enterococcus faecium strains isolated from ewe milk and artisanal cheese from northwest Argentina were screened for biotechnological properties relevant to flavour development. The API ZYM test showed absence of proteases, presence of high amounts of peptidases, and high esterase-lipase activities. Low extracellular proteolytic activity was observed. Most strains produced diacetyl in milk, with E. faecium OvL 214 and OvL 254 being the best producers. Biomass and growth rate increased when citrate was added to the medium, suggesting that these strains could use citrate as a main energy source. After 24 h of incubation, citrate was completely consumed in complex medium supplemented with glucose and citrate. An average of 17% residual citrate was detected in complex media supplemented with citrate. For all strains, esterase activity was detected up to alpha-naphthyl-caproate. They hydrolyzed alpha-naphthyl derivatives of fatty acids in this order: C3 > C6 > C4 > C8 > C2. Post-electrophoretic detection of esterase activities revealed the presence of multiple esterases. Hydrolysis of tributyrin, tricaprylin, and milk fat was observed in cell-free extracts. Enterococcus faecium strains isolated from ewe milk and artisanal cheese from northwest Argentina present the metabolic potential to contribute to cheese flavour development.

Glycerol metabolism of Lactobacillus rhamnosus ATCC 7469: cloning and expression of two glycerol kinase genes.

Lactobacillus rhamnosus ATCC 7469 was able to grow in glycerol as the sole source of energy in aerobic conditions, producing lactate, acetate, and diacetyl. A biphasic growth was observed in the presence of glucose. In this condition, glycerol consumption began after glucose was exhausted from the culture medium. Glycerol kinase activity was detected in L. rhamnosus ATCC 7469, a characteristic of microorganisms which catabolize glycerol in aerobic conditions. Genetic analysis revealed that this strain possesses two glycerol kinase genes: gykA and glpK, that encode for two different glycerol kinases GykA and GlpK, respectively. The glpK geneis associated in an operon with alpha-glycerophosphate oxidase (glpO) and glycerol facilitator (glpF) genes. Transcriptional analysis revealed that only glpK is expressed when L. rhamnosus was grown on glycerol.

Differentiation of lactic acid bacteria strains by postelectrophoretic detection of esterases.

Lactic acid bacteria (LAB) comprise a diverse group of Gram-positive, non-spore-forming microorganisms. These bacteria are widely used in food technology. The species identification of LAB depends mainly on physiological and biochemical criteria. The esterolytic systems of LAB remain poorly characterized. Esterases (EC 3.1.1.3) represent a diverse group of hydrolases catalyzing the cleavage and formation of esters bonds Screening of esterases is usually performed either by employing chromophoric substances (e.g., alpha- or beta-naphthyl esters of short-chain fatty acids). The post-electrophoretic detection of esterases is a sensitive technique applied in bacterial systems, that mainly provides information on the similarity of strains within the same species or subspecies according to their esterase patterns. This technique is principally used to determine the number and substrate specificity of esterases and lipases, revealing the complexity of lipase and esterase systems. The present chapter describes the technique of polyacrylamide gel electrophoresis (PAGE; in the absence of sodium dodecyl sulfate [SDS]), in non-denaturing conditions, to find intracellular fractions for strain typing of LAB.