Development and characterization of an innovative synbiotic fermented beverage based on vegetable soybean

Abstract Soymilk was produced from vegetable soybean and fermented by probiotics (Lactobacillus acidophilus La-5, Bifidobacterium animalis Bb-12) in co-culture with Streptococcus thermophilus. The composition of the fermented beverage and oligosaccharides content were determined. The effect of fructooligosaccharides and inulin on the fermentation time and viability of probiotic microorganisms throughout 28 days of storage at 5 °C were evaluated. The soymilk from vegetable soybeans was fermented in just 3.2 h, when pH reached 4.8. Fermentation reduced the contents of stachyose and raffinose in soymilk. Prebiotics had no effect on acidification rate and on viability of B. animalis and S. thermophilus in the fermented beverage. The viable counts of B. animalis Bb-12 remained above 108 CFU mL-1 in the fermented soymilk during 28 days of storage at 5 °C while L. acidophilus La-5 was decreased by 1 log CFU mL-1. The fermented soymilk from vegetable soybeans showed to be a good food matrix to deliver probiotic bacteria, as well as a soy product with a lower content of non-digestible oligosaccharides.

Influencia del aceite esencial de orégano en la elaboración tradicional de quesos: efecto sobre el fermento láctico / Influence of oregano essential oil on traditional Argentinean cheese elaboration: Effect on lactic starter cultures

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 differences 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 (enterobacteria 2 × 10³ UFC/g) and cheese flavored with oregano leaves (mold/yeast 4 × 10(4) CFU/g). Our results showed that the use of oregano essential oil and lactic starter culture considerably improved cheese quality.

El objetivo de este trabajo fue estudiar la composición del aceite esencial de orégano recolectado en el noroeste argentino y evaluar su efecto sobre algunos fermentos lácticos. El orégano recolectado correspondió a la especie Origanum vulgare var. hirtum proveniente de Andalgalá, Catamarca. En su aceite esencial (obtenido por arrastre con vapor de agua) se detectó principalmente α-terpineno (10%), γ-terpineno (15,1%), terpinen-4-ol (15,5%) y timol (13,0%). El aceite esencial no tuvo efecto inhibitorio (máxima concentración ensayada 200 μg/g) sobre el crecimiento ni sobre la actividad metabólica de Streptococcus thermophilus CRL 728 y CRL 813, de Lactobacillus delbrueckii subsp. bulgaricus CRL 656 y CRL 468, y de Lactococcus lactis subsp. lactis CRL 597. No se observaron diferencias en las características organolépticas de quesos semiduros aromatizados con el aceite esencial (200 μg/g) comparados con quesos artesanales aromatizados con hojas de orégano. Respecto de la calidad microbiológica de los productos, no se detectaron enterobacterias ni hongos o levaduras durante la maduración en los quesos aromatizados con el aceite esencial de orégano comparados con los quesos control, que presentaron desarrollo de enterobacterias (2 × 10³ UFC/g), y con los quesos elaborados con hojas de orégano, en los que hubo desarrollo de hongos/levaduras (4 × 10(4) CFU/g). Los resultados obtenidos demostraron que el uso del aceite esencial de orégano y del fermento láctico incrementó la calidad general de los quesos artesanales.

Efeito da composição de bactérias láticas e da suplementação do soro de leite na cinética de acidificação, crescimento celular e produção de nisina / Effect of lactic acid bacteria composition and the supplementation of milk whey on acidification kinetics, cell growth and nisin production

As bactérias ácido-láticas (BAL) são micro-organismos que auxiliam nas características organolépticas, funcionais e de bioconservação de produtos fermentados. A utilização do soro de leite como meio de cultivo natural enaltece o conceito da produção de biomoléculas de alto valor agregado, como bacteriocinas, já que é um subproduto gerado por indústrias de laticínios e considerado um agente poluidor. A inulina é um ingrediente prebiótico que promove seletivamente o crescimento de culturas probióticas. Nesse âmbito, o objetivo deste estudo foi avaliar o efeito da composição da cultura de Lactococcus lactis (LL) em cocultura com Streptococcus thermophilus (ST) e da suplementação da base de soro de leite com inulina: (i) nos parâmetros cinéticos de acidificacão, (ii) no crescimento celular, (iii) na viscosidade do produto e (iv) na atividade antimicrobiana da nisina. A fermentação do soro de leite com Lactococcus lactis em cocultura com Streptococcus thermophilus proporcionou a maior taxa de acidificação (Vmax=7,93x10-3 upH/min), assim como apresentou o menor tempo para atingir a velocidade máxima de acidificação (Tvmax=1,13 h). A adição de 2% de inulina ao soro de leite fermentado pela cocultura binária fez com que o tempo para completar o cultivo fosse o mais curto (TpH4,5=4,43 h) quando comparado aos demais ensaios. Quanto ao crescimento celular, pode-se observar que a inulina não afetou significativamente a contagem microbiológica, quando as cepas ST e LL foram utilizadas separadamente no processo fermentativo. Em particular, a adição de 4% de inulina reduziu em 1,2 LogUFC/mL e 0,92 LogUFC/mL a contagem de ST e LL (em monocultura), respectivamente. Por outro lado, em coculturas binárias (ST-LL), percebeu-se ganho na contagem microbiológica nos ensaios que receberam suplementação do ingrediente prebiótico, ou seja, quando adicionados 2% e 4% de inulina, houve aumento de 1 LogUFC/mL e de 1,34 LogUFC/mL na contagem de ST, respectivamente. No caso da cepa LL em cocultura com ST, a suplementação de 2% e 4% do prebiótico aumentou em 0,31 LogUFC/mL e 0,75 LogUFC/mL, respectivamente. A concentração de ácido lático também foi mais elevada nos cultivos realizados com a cocultura binária, sendo 4,56 g/L (na ausência de inulina), 5,28 g/L (com adição de 2% de inulina) e 5,71 g/L (com suplementação de 4% de inulina). A viscosidade foi influenciada tanto pela adição de inulina como pelo efeito sinérgico da cocultura, sendo que o maior valor (7,38 mPas) foi obtido pela cocultura ST-LL e pela adição de 4% do ingrediente prebiótico. Quanto à produção de nisina, observou-se que, no cultivo em cocultura (ST-LL), a concentração de 2% de inulina aumentou em 102% a atividade antimicrobiana quando comparada com a cultura pura LL. Vale ressaltar que ambas as cepas satisfizeram os requisitos tecnológicos relativos à produção de laticínios funcionais

Lactic acid bacteria (LAB) are microorganisms that help in the organoleptic and functional characteristics and in the biopreservation of fermented products. The use of milk whey as a culture medium extols the concept of the production of high value-added biomolecules, such as bacteriocins, since it is a by-product generated by the dairy industry and considered a pollutant. Inulin is a prebiotic ingredient that promotes selectively the growth of probiotic cultures. In this context, the aim of this study was to evaluate the effect of culture composition Lactococcus lactis (LL) in co-culture with Streptococcus thermophilus (ST) and the supplementation of milk whey with inulin on: (i) the acidification kinetic parameters, (ii) the cell growth, (iii) the product viscosity, and (iv) the antimicrobial activity of nisin. The fermentation of milk whey by Lactococcus lactis in coculture with Streptococcus thermophilus provided the highest acidification rate (Vmax = 7.93x10-3 upH/min) and the shortest time to reach the maximum acidification rate ( TVmax = 1.13 h). The addition of 2% inulin in the binary coculture binary led to the shorter time to complete the fermentation (TpH4,5 = 4.43) compared to the other tests. With regard to cell growth, it can be observed that the addition of inulin did not affect the microbiological count of pure cultures of ST and LL strains in the fermentation process. In particular, the addition of 4% inulin reduced by 1.2 Log CFU/mL and 0.92 Log CFU/mL the counts of ST and LL (monoculture), respectively. In the other hand, the binary co-cultures cultivations (ST-LL) with the addition of 2% and 4% inulin increased by 1 LogCFU/mL and 1.34 Log CFU/mL in the case of the ST counts and 0.31 log CFU/mL and 0.75 log CFU/mL the counts of LL, respectively. Lactic acid concentration was higher in cultivations carried out by binary cocultures, thus being 4.56 g/L (in the absence of inulin), 5.28 g/L (with addition of 2% inulin) and 5.71g/L (supplemented with 4% inulin). The viscosity was influenced by the addition of prebiotic ingredient and by the synergistic effect of binary coculture, being the highest value (7.38 mPas) obtained by the addition of 4% inulin. Finally, as regards the production of nisin noted that in the binary coculture cultivations (ST-LL), the concentration of 2% inulin increased at 102% the antimicrobial activity when compared to the pure culture LL. It is worth mentioning that both strains met the technological requirements as regards the production of functional dairy products

Comportamento de cepas distintas de Lactobacillus acidophilus em queijo petit-suisse

O objetivo do presente trabalho foi avaliar as características de físico-químicas e microbiológicas de queijo petit-suisse processado com a adição de duas cepas de Lactobacillus acidophilus: LA-14 (potencialmente probiótica) e La-5 (comprovadamente probiótica), utilizando Streptococcus thermophilus TA040 como cultura starter. Três queijos petit-suisse foram preparados: Q1 (controle:TA040), Q2 (TA040 + LA-14) e Q3 (TA040 +La-5). Foram realizadas análises microbiológicas (determinação das populações dos microrganismos La-5, La-14 e TA040) e físico-químicas (umidade e pH) após 1, 7, 14, 21 e 28 dias de armazenamento dos produtos a 4±1°C. As populações de L. acidophilus oscilaram entre 7,46 e 7,62 log UFC g -1 para La-5 e entre 6,39 e 6,83 log UFC g -1 para LA-14, evidenciando que a sobrevivência de L. acidophilus no produto depende de características particulares da cepa. Populações superiores da cultura starter foram observadas para Q2 (9,58 - 9,68 log UFC g -1 ) e Q3 (9,42 - 9,79 log UFC g -1 ), quando comparadas a Q1 (9,11 - 9,23 log UFC g -1 ), sugerindo sinergismo entre L. acidophilus e o starter. A umidade e o pH permaneceram estáveis e não diferiram entre os queijos após o 1º dia de armazenamento (p>0,05). As características peculiares das cepas de L. acidophilus determinaram os comportamentos distintos observados nos queijos petitsuisse, sendo possível detectar a melhor adaptação da cepa La-5 ao produto, o que resultou em populações significativamente superiores quando comparada a LA-14.

Particular behavior of different Lactobacillus acidophilus strains in petit-suisse cheese. The objective of this study was to evaluate the physico-chemical andmicrobiological characteristics of petit-suisse cheeses manufactured with the addition of two Lactobacillus acidophilus strains: LA-14 (potentially probiotic) and La-5 (probiotic culture), using Streptococcus thermophilus TA040 as starter culture. Three cheese-making trials were prepared: Q1 (control: with TA040), Q2 (with TA040 + LA-14), and Q3 (with TA040 + La-5). Parameters analyzed included microbial counts of probiotic, potentially probiotic and starter microorganisms, and physico-chemical parameters (pH and moisture) after 1, 7, 14, 21, and 28 days of storage of the product at 4±1°C. Viable counts of L. acidophilus remained between 7.46 and 7.62 log CFU g -1 for La-5, and between 6.39 and 6.83 log CFU g -1 for LA-14. As for the starter, higher populations were observed for Q2 (9.58 - 9.68 log CFU g -1 ) and Q3 (9.42 - 9.79 log CFU g -1 ), when compared to Q1, which suggests synergism between L. acidophilus and the starter culture. Moisture and pH values remained stable for cheeses Q1, Q2, and Q3, and no significant differences were detected between cheeses after the first day of storage (p>0.05). Particular features of both L. acidophilus strains determined different behavior in petit-suisse cheese, and the better adaptation of the La-5 to the product environment was perceptible, since higher populations were observed when compared to LA-14.

Growth and exopolysaccharide production by Streptococcus thermophilus ST1 in skim milk

To analyze the exopolysaccharide (EPS) production by Streptococcus thermophilus ST1, cultures were cultivated in 10 percent (w/v) reconstituted skim milk under different growth conditions including various temperatures and pHs of growth medium, supplementation of the medium with various carbon sources (glucose, lactose, sucrose, galactose and fructose) and nitrogen source (whey protein concentrate, or WPC). The results showed that most EPS production by strain ST1 was obtained at a temperature (42°C) and pH (6.5) optimal for its growth. Supplementation of the skim milk medium with either carbohydrates or WPC increased both growth and polymer formation by different extents, with sucrose being most effective among the carbon sources tested. Under the optimal cultural conditions, i.e. pH 6.5, 42°C with 2 percent (w/v) sucrose and 0.5 percent (w/v) WPC, 135.80 mg l-1 of EPS was produced by strain ST1. The monosaccharide composition of the EPS was determined to be glucose and galactose (2:1), and the molecular mass of the EPS was 3.97 × 10(6) Da. The aqueous solution of the EPS at 1 percent (w/v) showed relatively high viscosity, indicating the potential of this EPS-producing S. thermophilus strain for applications in the improvement of physical properties of fermented milk products.