Quantification of the environmental impact arising from the utilization of whole and defatted Okara in fermentative and dehydration processes.

Okara is the insoluble pulp that remains after the grinding and filtration of soybeans during the production of soymilk and tofu. As it retains a noteworthy quantity of nutrients, there has been an increasing emphasis in the utilization of this residue for the development of sustainable processes. This study focused on assessing the environmental impact of employing okara as a medium for fermenting and dehydrating probiotic bacteria at laboratory scale. The evaluation was carried out using the Life Cycle Assessment (LCA) methodology, considering the entire process lifecycle. Whole okara and defatted okara were used as culture media for Lactiplantibacillus plantarum CIDCA 83114, followed by dehydration (either freeze-drying or spray-drying) and subsequent storage. For the purpose of comparison, both scenarios (whole and defatted okara) were evaluated using 1 kg of dehydrated final product for storage, as functional unit. Based on experimental results, the conservation of the received okara and the dehydration-storage (e.g., freezing and freeze-drying) phases were identified as the most significant environmental hotspots responsible for the most substantial impacts of the processes. The use of LCA facilitated the measurement of the environmental effects linked to the reutilization of okara as an agro-industrial residue, thus providing quantitative support when engineering its sustainable valorization.

Profile and content of isoflavones on flaked and extruded soybeans and okara submitted to different drying methods.

Isoflavones (IFs) are biocompounds found in considerable amounts in soybean grains. However, to originate soybean-based materials, the grains must be subjected to numerous thermal and mechanical treatments, which can impair the IFs content. The influence of these treatments was investigated and a protocol for IFs extraction and quantification is proposed. Sequential extractions were performed on industrially pretreated soybean samples (broken, flakes, and collets), on okara submitted to different drying methods (freeze-drying, forced convection, and under vacuum), and on soybean oils extracted with hexane and ethanol. ß-glucosides levels were decreased by the thermal processes of lamination, expansion, and drying, while the aglycone forms were not affected. Lyophilization was identified as the most viable drying method for the quantification of IFs in okara. Soybean oils extracted with ethanol presented significant amounts of aglycone. Furthermore, two stages of extraction were sufficient for the recovery of the IFs from different matrices.

Microencapsulation of Lactobacillus plantarum in W/O emulsions of okara oil and block-copolymers of poly(acrylic acid) and pluronic using microfluidic devices.

Okara oil is a by-product remaining from defatting okara, the solid residue generated after extracting the aqueous fraction of grounded soybeans in the elaboration of soy beverages. The goal of this work was to encapsulate the probiotic Lactobacillus plantarum CIDCA 83114 into W/O emulsions composed of a block-copolymer constituted of pluronic® and acrylic acid (PPP12) and okara oil, prepared in microfluidic devices. For comparative purposes, alginate was also included as a second dispersed phase. Lactobacillus plantarum CIDCA 83114 was suspended in PPP12 or alginate giving rise to dispersed phases with different compositions, named I, II, III and IV. Controls were prepared by suspending microorganisms in water as dispersed phase. 6-carboxyfluorescein was added as bacterial marker in all the emulsions. The presence of green dyed bacteria in the dispersed phases, inside the droplets of the emulsions and the absence of fluorescence outside them, confirmed the complete encapsulation of bacteria in the dispersed phases. After being prepared, emulsions were freeze-dried. The exposure to gastric conditions did not lead to significant differences among the emulsions containing polymers. However, in all cases bacterial counts were significantly lower than those of the control. After exposing emulsions to the simulated intestinal environment, bacterial counts in assays I, II and III (emulsions composed of only one dispersed phase or of two dispersed phases with bacteria resuspended in the PPP12 one) were significantly greater than those of the control (p < 0.05) and no detectable microorganisms were observed for assay IV (emulsions composed of two dispersed phases with bacteria resuspended in the alginate one). In particular, bacterial cultivability in emulsions corresponding to assay I (only PPP12 as dispersed phase) exposed to the intestinal environment was 8.22 ± 0.02 log CFU/mL (2 log CFU higher than the values obtained after gastric digestion). These results support the role of PPP12 as an adequate co-polymer to protect probiotics from the gastric environment, enabling their release in the gut, with great potential for food or nutraceutical applications.

Effect of different gluten-free flours on the sensory characteristics of a vegan alfajor: Vegan gluten-free Alfajor development.

Vegan and gluten-free markets have grown considerably in the last few years. Sustainability and the use of agro-industrial waste have also gained interest on food market. Thus, this study aimed to develop a vegan gluten-free alfajor, assessing the effect of different gluten-free flours (peanut okara (a by-product), sorghum, and rice flours) on the product sensory profile, and its market appeal. A simplex centroid design was applied to optimize the alfajor formulation. Check All That Apply and acceptance tests were performed. The use of different flours and their mixtures generated products with different texture attributes. All formulations obtained good acceptances, but higher concentrations of peanut okara and sorghum flours contributed to produce softer alfajors, considered as preferred by consumers. Therefore, it was possible to develop a gluten-free alfajor with sensory quality adding value to an agroindustrial by-product.

Enzymatic pretreatment in the extraction process of soybean to improve protein and isoflavone recovery and to favor aglycone formation.

This research aimed to evaluate the pretreatment of soybean with the carbohydrase multi-enzyme complex "Viscozyme L", during the extraction process; in order to improve the recovery of proteins and isoflavones in soybase, and reduce the loss of these compounds through the okara residue. Three concentrations of enzyme were studied at 50 °C, along with an experiment carried out without enzyme addition (control experiment). The results were also evaluated in relation to standard soybase processing. In comparison to the standard and control processes, the enzymatic pretreatment reduced up to 85% the total amount of okara residue. Due to the action of the multi-enzyme complex, protein and total isoflavone recovery increased from 42% to 83% and from 59% to 93%, respectively. The application of Viscozyme L also favored the conversion of conjugated forms of isoflavone to aglycone in the soybase, representing up to 50% of the total isoflavones. The enzymatic pretreatment of soybean with carbohydrase improved the nutritional quality of the soybase, while at the same time reducing residue generation; showing that the proposed food process can be considered environmentally friendly method.

Release of health-related compounds during in vitro gastro-intestinal digestion of okara and okara fermented with Lactobacillus plantarum.

Okara is a highly perishable by-product remaining after filtration of the smashed soybeans seeds in the production of soymilk. Due to its nutritional value, different approaches have been developed to use it as functional ingredient. Fermentation of okara appears as an interesting strategy to preclude spoilage, providing a more stable matrix to be incorporated in the formulation of functional foods. Okara has antioxidant compounds but the effect of fermentation, and their bioaccessibility still need to be investigated. To achieve this aim, the phenolic compounds (as determined by TPC and TFC assays) and the antioxidant properties (as determined by ABTS ·+, DPPH · , O2 ·- assays) of okara and okara fermented with Lactobacillus plantarum CIDCA 83114 were assessed both before and after exposure to simulated gastro-intestinal conditions. Before digestion, okara showed higher values of TPC and TFC than the fermented counterpart. Although a decrease of TPC and TFC was observed after exposing okara to gastric conditions, no significant differences between okara and fermented okara were detected. No further decrease of TPC were observed in intestinal conditions. Okara showed higher antioxidant activity than fermented okara. There was a considerable decrease in the antioxidant activity for both samples when exposed to gastric and intestinal conditions. A good correlation between TFC and antioxidant activities was detected, suggesting that flavonoids play an important role as antioxidants. As a whole, this work provides a solid support for the stability of phytochemicals along the digestive process of both okara and fermented okara.

Aflatoxins distribution in fractions derived from tofu production.

Tofu or bean curd is obtained from soybean seeds being a widespread food product in Asia. The commodity used for its production can be contaminated with aflatoxins, which are secondary metabolites synthetised by species of Aspergillus flavus and A. parasiticus. Intake of contaminated food products causes toxic effects on consumers. The aim of this work was to study aflatoxin distribution in fractions obtained from pilot-scale tofu production with contaminated soybeans. The presence of the aflatoxins B1, B2, G1 and G2 (AFs) in soaking water, okara, whey and tofu was analysed. Aflatoxin analysis was performed by HPLC with fluorescence detection. The distribution of aflatoxins in all the analysed fractions was not a normal distribution. The liquid fractions (soaking water and whey) had less contamination than solid fractions (tofu and okara). The percentage AFB1 remaining in nutritionally important fractions, okara and tofu, was between 6.2% and 67.7% (median = 18.1%) and 0.5% and 13.2% (median = 3.5%), respectively. AFB2, AFG1 and AFG2 had a similar distribution. These results showed that throughout tofu production, AFs can be present in the products intended for human consumption.

The impact of fruit and soybean by-products and amaranth on the growth of probiotic and starter microorganisms.

The ability of different fruit by-products, okara, and amaranth flour, to support the growth of probiotic and non-probiotic strains was evaluated. The tests were conducted with three commercial starter cultures (Streptococcus thermophilus), ten probiotic strains (seven Lactobacillus spp. and three Bifidobacterium spp. strains), and two harmful bacteria representative of the intestinal microbiota (Escherichia coli and Clostridium perfringens). In vitro fermentability assays were performed using a modified MRS broth supplemented with different fruits (acerola, orange, passion fruit, and mango), and soy (okara) by-products or amaranth flour. Orange and passion-fruit by-products were the substrates that most promoted the growth of bacterial populations, including pathogenic strains. On the other hand, the acerola by-product was the substrate that showed the highest selectivity for beneficial bacteria, since the E. coli and Cl. perfringens populations were lower in the presence of this fruit by-product. Although the passion fruit by-product, okara, and amaranth stimulated the probiotic strains, the growth of the pathogenic strains studied was higher compared to other substrates. Different growth profiles were verified for each substrate when the different strains were compared. Although pure culture models do not reflect bacterial interaction in the host, this study reinforces the fact that the ability to metabolize different substrates is strain-dependent, and acerola, mango, and orange by-products are the substrates with the greatest potential to be used as prebiotic ingredients.

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage.

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum. Considering that okara is an agro-waste obtained in large quantities, these results represent an innovative strategy to add it value, providing a symbiotic ingredient with promising industrial applications in the development of novel functional foods and feeds.

Supplementation with fruit and okara soybean by-products and amaranth flour increases the folate production by starter and probiotic cultures.

The ability of two starter cultures (Streptococcus (S.) thermophilus ST-M6 and St. thermophilus TA-40) and eleven probiotic cultures (St. thermophilus TH-4, Lactobacillus (Lb.) acidophilus LA-5, Lb. fermentum PCC, Lb. reuteri RC-14, Lb. paracasei subsp. paracasei, Lb. casei 431, Lb. paracasei subsp. paracasei F19, Lb. rhamnosus GR-1, and Lb. rhamnosus LGG, Bifidobacterium (B.) animalis subsp. lactis BB-12, B. longum subsp. longum BB-46, and B. longum subsp. infantis BB-02) to produce folate in a modified MRS broth (mMRS) supplemented with different fruit (passion fruit, acerola, orange, and mango) and okara soybean by-products and amaranth flour was investigated. Initially, the folate content of each vegetable substrate was determined: passion fruit by-product showed the lowest folate content (8±2ng/mL) and okara the highest (457±22ng/mL). When the orange by-product and amaranth flour were added to mMRS, all strains were able to increase folate production after 24h of fermentation. B. longum subsp infantis BB-02 produced the highest concentrations (1223±116ng/mL) in amaranth flour. Okara was the substrate that had the lowest impact on the folate production by all strains evaluated. Lb. acidophilus LA-5 (297±36ng/mL) and B. animalis subsp. lactis BB-12 (237±23ng/mL) were also able to produce folate after growth in mMRS containing acerola and orange by-products, respectively. The results of this study demonstrate that folate production is not only strain-dependent but also influenced by the addition of different substrates in the growth media.

Nutritional recovery with okara diet prevented hypercholesterolemia, hepatic steatosis and glucose intolerance.

We assessed the biological value of an okara diet and its effects on the hormonal and metabolic profile of rats submitted to protein restriction during intra-uterine life and lactation and recovered after weaning. Male rats from mothers fed either 17% or 6% protein during pregnancy and lactation were maintained on 17% casein (CC, LC), 17% okara (CO, LO) or 6% casein (LL) diets over 60 d. The nutritional quality of the okara protein was similar to that of casein. The okara diet was effective in the nutritional recovery of rats in growing that were malnourished in early life. Furthermore, the okara diet reversed the hypercholesterolemia and the hepatic steatosis observed in the malnutrition and prevented glucose intolerance in an animal model prone to diabetes mellitus.

Incorporation of soybean by-product okara and inulin in a probiotic soy yoghurt: texture profile and sensory acceptance.

BACKGROUND: This study evaluated the effect of inulin and okara flour on textural and sensory properties of probiotic soy yoghurt (SY) throughout 28 days of storage at 4 °C. Employing a 2(2) design, four formulations of SY produced from soymilk and fermented with an ABT-4 culture (Lactobacillus acidophilus La-5, Bifidobacterium animalis Bb-12 and Streptococcus thermophilus) were studied: SY-C (control); SY-I (with inulin); SY-O (with okara); SY-IO (with inulin + okara). RESULTS: The addition of okara and the refrigerated storage led to significant differences in the instrumental texture parameters of SY (P < 0.05). Inulin and okara did not affect SY sensory acceptability (P > 0.05), but there was a tendency for higher scores in the presence of inulin. On the other hand, the storage period, particularly at 21 days, was unfavourable regarding the acceptance of the different SY. CONCLUSION: The results showed that the addition of okara flour and the storage were significant factors to increase firmness of the soy yoghurts. SY acceptability was not affected by the incorporation of inulin or okara. These results suggest that okara, discarded as industrial waste, may be used in probiotic soy yoghurt, helping to increase the nutritional and functional properties without altering its acceptability.

Análises físico-químicas de pré-misturas de pães de queijo e produção de pães de queijo com adição de okara / Physicochemical analyses of commercial samples of cheese bread premix and production of cheese breads with addition of okara

Okara, ou resíduo de soja, é um subproduto do processamento do extrato aquoso de soja e do tofu. Pão de queijo é um produto assado ao forno obtido a partir da mistura de polvilho com água ou leite, queijo, sal e gordura, podendo ser utilizados polvilho doce, azedo ou a mistura deles. Objetivou-se com este trabalho caracterizar pré-misturas comerciais de pão de queijo por intermédio de análises físico-químicas e também avaliar a qualidade de pães de queijo produzidos com a adição de subproduto da obtenção do extrato aquoso de soja. As características de qualidade de pães de queijo suplementados com 5, 10 e 15 por cento de okara foram investigadas. Os pães de queijo suplementados com subproduto okara apresentaram teores de proteínas e de fibras alimentares superiores ao controle. Os produtos panificados foram submetidos à análise sensorial de aceitabilidade utilizando-se a escala hedônica de nove pontos, com provadores não-treinados. Amostras de pães de queijo com 5, 10 e 15 por cento de okara não foram consideradas diferentes significativamente em nível de 5 por cento e tiveram boa aceitação.

Okara, or soy residue, is a byproduct of soy aqueous extract and tofu manufacturing. Cheese bread is a Brazilian specialty made by blending cassava starch (cassava starch or sour cassava starch) water or milk, cheese, salt and fat and is baked on oven. This study has the objective of characterizing cheese breads made with addition of okara. Additionally in this work commercial samples of cheese bread premix were compared in terms of physicochemical properties. The quality characteristics of cheese breads supplemented with 5, 10 and 15 percent of okara were investigated. The results showed that the cheese breads supplemented with the byproduct okara had an increase in the protein and dietary fiber contents. The samples were submitted to an acceptability sensory evaluation with a nine point hedonic scale, involving untrained panelists. The cheese breads made with 5, 10 and 15 percent of okara were not statistically different at the level 5 percent and had good acceptability.