RESUMEN
Food allergies represent a serious problem affecting human health and soy proteins rank among the most allergenic proteins from food origin. The proteolytic enzymes produced by lactic acid bacteria (LAB) can hydrolyse the major allergens present in soybean, reducing their immunoreactivity. Many studies have reported the ability of LAB to ferment soy-based products; while the majority of them focus on the improvement of the sensory characteristics and functionality of soy proteins, a lack of information about the role of lactic fermentation in the reduction of immunoreactivity of these proteins exists. The aim of the present study was to evaluate the capability of the proteolytic strain Enterococcus faecalis VB43 to hydrolyse the main allergenic proteins present in soymilk and to determine the immunoreactivity of the obtained hydrolysates. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results of fermented soymilk demonstrated complete hydrolysis of the ß-subunit from ß-conglycinin and the acidic polypeptide from glycinin. Reversed phase high performance liquid chromatography (RP-HPLC) analysis of the peptides released after hydrolysis revealed the appearance of new peptides and the disappearance of non-hydrolysed proteins, indicating extensive hydrolysis of the substrate. Results from competitive enzyme-linked immunosorbent assay (ELISA) tests clearly indicated a reduction in the immunoreactivity (more than one logarithmic unit) in the fermented sample as compared to the non-fermented control. Our results suggest that the soymilk fermented by E. faecalis VB43 may induce lower allergic responses in sensitive individuals. The strain E. faecalis VB43 may be considered as an excellent candidate to efficiently reduce the immunoreactivity of soymilk proteins.
Asunto(s)
Antígenos de Plantas/inmunología , Enterococcus faecalis/metabolismo , Globulinas/inmunología , Proteínas de Almacenamiento de Semillas/inmunología , Leche de Soja/metabolismo , Proteínas de Soja/inmunología , Antígenos de Plantas/química , Antígenos de Plantas/metabolismo , Cromatografía Líquida de Alta Presión , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Fermentación , Globulinas/química , Globulinas/metabolismo , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/metabolismo , Leche de Soja/química , Proteínas de Soja/química , Proteínas de Soja/metabolismo , Glycine max/química , Glycine max/inmunología , Glycine max/metabolismo , Glycine max/microbiologíaRESUMEN
The use of solid fermentation substrate (SSF) has been appreciated by the demand for natural and healthy products. Lactic acid bacteria and bifidobacteria play a leading role in the production of novel functional foods and their behavior is practically unknown in these systems. Soy is an excellent substrate for the production of functional foods for their low cost and nutritional value. The aim of this work was to optimize different parameters involved in solid state fermentation (SSF) using selected lactic cultures to improve soybean substrate as a possible strategy for the elaboration of new soy food with enhanced functional and nutritional properties. Soy flour and selected lactic cultures were used under different conditions to optimize the soy SSF. The measured responses were bacterial growth, free amino acids and ß-glucosidase activity, which were analyzed by applying response surface methodology. Based on the proposed statistical model, different fermentation conditions were raised by varying the moisture content (50-80%) of the soy substrate and temperature of incubation (31-43°C). The effect of inoculum amount was also investigated. These studies demonstrated the ability of selected strains (Lactobacillus paracasei subsp. paracasei and Bifidobacterium longum) to grow with strain-dependent behavior on the SSF system. ß-Glucosidase activity was evident in both strains and L. paracasei subsp. paracasei was able to increase the free amino acids at the end of fermentation under assayed conditions. The used statistical model has allowed the optimization of fermentation parameters on soy SSF by selected lactic strains. Besides, the possibility to work with lower initial bacterial amounts to obtain results with significant technological impact was demonstrated.
Asunto(s)
Bifidobacterium/metabolismo , Fermentación , Microbiología de Alimentos , Glycine max/microbiología , Lactobacillus/metabolismo , Alimentos Funcionales/microbiología , Lactobacillaceae/metabolismo , Proteínas de Soja/metabolismoRESUMEN
AIMS: Consumption of soya-derived products has been hampered by the presence of alpha-galactooligosaccharides (alpha-GOS) because mammals lack pancreatic alpha-galactosidase (alpha-Gal) which is necessary for their hydrolysis. These sugars thus reach the large intestine causing gastrointestinal disorders in sensitive individuals. The use of lactic acid bacteria (LAB) expressing alpha-Gal is a promising solution for the degradation of alpha-GOS in soyamilk. METHODS AND RESULTS: The capacity of the LAB Lactobacillus fermentum CRL 722 to properly degrade alpha-GOS was studied in vitro using controlled fermentation conditions and in vivo using a rat model. Lactobacillus fermentum CRL 722 was able to grow on commercial soyamilk and completely eliminated stachyose and raffinose during fermentation because of its high alpha-Gal activity. Rats fed soyamilk fermented by this LAB had smaller caecums compared with rats fed unfermented soyamilk. CONCLUSIONS: Soyamilk fermentation by Lact. fermentum CRL 722 results in the reduction of alpha-GOS concentrations in soyamilk, thus eliminating possible undesirable physiological effects normally associated with its consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: Fermentation with Lact. fermentum CRL 722 could prevent gastrointestinal disorders in sensitive individuals normally associated with the consumption of soya-based products. This LAB could thus be used in the elaboration of novel fermented vegetable products which better suit the digestive capacities of consumers.
Asunto(s)
Microbiología de Alimentos , Galactosa/metabolismo , Lactobacillus/metabolismo , Oligosacáridos/metabolismo , Leche de Soja , Animales , Ciego/anatomía & histología , Ciego/metabolismo , Recuento de Colonia Microbiana/métodos , Fermentación , Lactobacillus/crecimiento & desarrollo , Rafinosa/metabolismo , Ratas , Ratas Wistar , alfa-Galactosidasa/metabolismoRESUMEN
In this study, the behaviour of Lactobacillus fermentum CRL 722 and CRL 251 were evaluated under different pH conditions (pH 6.0, 5.5, 5.0, 4.5) and without pH control. Growth was similar under all conditions assayed except at pH 4.5. These microorganisms were able to eliminate raffinose, a nondigestible alpha-oligosaccharide (NDO) found in soy products, showing a consumption rate of 0.25 g l(-1) h(-1) (pH 6.0-5.0). The removal of raffinose was due to the high alpha-galactosidase (alpha-gal) activities of these lactic acid bacteria, which was highest at pH 5.5 (5.0 U/ml). The yield of organic acids produced during raffinose consumption was also highest at this pH. The results of this study will allow selection of the optimum growth conditions of L. fermentum with elevated levels of alpha-gal to be used in the reduction of NDO in soy products when used as starter cultures.
Asunto(s)
Concentración de Iones de Hidrógeno , Lactobacillus/crecimiento & desarrollo , Lactobacillus/metabolismo , Rafinosa/metabolismo , alfa-Galactosidasa/metabolismo , Fermentación , Microbiología de Alimentos , Ácido Láctico/metabolismo , Lactobacillus/enzimologíaRESUMEN
The refrigerated shelf life of soymilk fermented with single cultures of Lactobacillus fermentum, L. casei, Streptococcus salivarius subsp. thermophilus, and Bifidobacterium longum was evaluated. During storage at 4 degrees C for 28 days, the stability of the microflora differed markedly among the starter cultures. After 28 days, the average numbers of S. salivarius subsp. thermophilus decreased by two log cycles to 6.0 x 10(7) CFU/ml, whereas those of L. casei increased gradually by more than two log cycles to 4.6 x 10(9) CFU/ml. Numbers of B. longum and L. fermentum remained moderately high (8.7 x 10(8) CFU/ml and 3.7 x 10(8) CFU/ml, respectively) even after 28 days of storage. S. salivarius subsp. thermophilus and L. casei continued to metabolize sucrose during the storage period, but the pattern of consumption was different among the strains. The other starter cultures did not seem to have significant activity (P > 0.05) on the residual sugars. In most cases, L(+)-lactate predominated.