Content composition and antioxidant activity of isoflavones in commercial and homemade soymilk and tofu

BACKGROUND: Isoflavones, found in soymilk and tofu, are one of the phytochemicals in soy‐based products that may promote good health. Homemade tofu and various homemade soymilk samples were made using different soaking, grinding, and cooking methods. The homemade samples were compared to commercial tofu and soymilk for total isoflavone content and composition as well as their antioxidant capacity. All samples were freeze‐dried and extracted with a 58% acetonitrile solution which was subsequently used to determine the isoflavone content by reverse‐phase high‐performance liquid chromatography. The antioxidant activity of extracts was determined using a modified 2,2‐azino‐bis‐(3‐ethylbenzthiazoline‐6‐sulfonic acid) method and total antioxidant capacity was reported as ascorbic acid equivalents. RESULTS: The total isoflavone, aglycone, and antioxidant levels were significantly higher in homemade soymilk and tofu (1571 µg) than in commercial samples. Homemade soymilk made by the extended boiling method yielded the highest total isoflavone (2567 µg) and glucoside (1525 µg) content. A strong positive correlation was observed between the total isoflavone, aglycone conjugates, and genistein series concentration and antioxidant capacity of soymilk. CONCLUSION: Increased moist heating time yielded the highest concentration of total isoflavones as well as aglycone conjugates and the genistein series. Increasing the duration of boiling can increase the isoflavone content of both homemade and commercial soymilk and tofu. Copyright © 2007 Society of Chemical Industry     

HYDROLYSIS OF ISOFLAVONE PHYTOESTROGENS IN SOYMILK FERMENTED BY LACTOBACILLUS AND BIFIDOBACTERIUM COCULTURES

This study evaluated the hydrolysis of isoflavones in soymilk fermented at 37C for 48 h by four different Lactobacillus and Bifidobacterium cocultures. The hydrolysis of isoflavone β-glucosides significantly increased ( P <  0.05) the bioactive aglycones from 36 to over 90% of total isoflavones in soymilk fermented with any of the four Lactobacillus and Bifidobacterium cocultures as compared with unfermented soymilk. Compared with three other cocultures of Lactobacillus and Bifidobacterium, fermentation of soymilk with the Lactobacillus paracasei/Bifidobacterium longum cocultures yielded better isoflavone hydrolytic potential (Otieno-Shah index) and the highest β-glucosidase activity after 12 h of incubation.

PRACTICAL APPLICATIONS

Isoflavones are known as phytoestrogens because they are present in soy products and have estrogen-like activity. During fermentation, the majority of glucoside isoflavones in soymilk are converted to bioactive aglycones via microorganism-derived β-glucosidase. In human intestines, aglycone isoflavones are absorbed faster and in greater amounts than their glucosides. Using probiotic Lactobacillus and Bifidobacterium cocultures to ferment soymilk efficiently increases the bioactive aglycone concentrations. Hence, fermenting soymilk with this coculture could enhance the nutritional value of the product.     

Soymilk fermented with Lactobacillus rhamnosus CRL981 ameliorates hyperglycemia,lipid profiles and increases antioxidant enzyme activities in diabetic mice

Interest in soybeans and soy-based products has grown significantly in the last decades due to their reported nutritional and health-promoting effects. In soybean and non-fermented soy foods, isoflavones are predominantly found as glucosides, which must be hydrolyzed in order to exert their documented beneficial effects. The objective of this study was to evaluate the effect of soymilk fermented with Lactobacillus rhamnosus CRL981, a strain that can completely hydrolyze glucoside isoflavones due to its high beta-glucosidase activity. Using a diabetes animal model induced with streptozotocin, it was shown that the fermented soymilk was able to significantly decrease glucose levels, total cholesterol concentrations, triacylglycerols and increase antioxidant enzyme activities compared to animals that received unfermented soymilk. This study clearly shows that the adequate selection of starter cultures could be used to develop novel soy based products intended for the prevention or adjuvant treatment of diseases such as diabetes.     

Isoflavone phytoestrogen degradation in fermented soymilk with selected beta-glucosidase producing L. acidophilus strains during storage at different temperatures

Soymilk fermented with 3 selected Lactobacillus acidophilus strains were stored at various temperatures (-80 degrees C, 4 degrees C, 25 degrees C and 37 degrees C) for 8 weeks and the concentration of isoflavones determined weekly using RP-HPLC. The decreasing concentration of isoflavones in soymilk during storage due to degradation was found to fit the first order kinetics model. Isoflavone aglycones as well as isoflavone glucosides largely appeared to be stable during storage (P<0.01). Interestingly, the aglycone forms showed much smaller degradation as compared to glucoside forms at all the storage temperatures studied. Of the isoflavone aglycones, daidzein was found to be the most stable followed by genistein, while glycitein was least stable. Isoflavone aglycones such as glycitein, daidzein and genistein showed smaller degradation constants in fermented soymilk at lower storage temperatures (-80 degrees C and 4 degrees C) and higher degradation constants at higher storage temperatures (25 degrees C and 37 degrees C) with each strain. In contrast, glucosides glycitin and daidzin showed higher degradation at lower storage temperatures (-80 degrees C and 4 degrees C) and lower degradation at higher storage temperatures (25 degrees C and 37 degrees C). Storage temperature was therefore found to be very important in regulating the rate of degradation soy isoflavones in fermented soymilk.     

Evaluation of enzymic potential for biotransformation of isoflavone phytoestrogen in soymilk by Bifidobacterium animalis,Lactobacillus acidophilus and Lactobacillus casei

Three strains of Lactobacillus acidophilus, two of Lactobacillus casei and one of Bifidobacterium were screened for β-glucosidase activity using ρ-nitrophenyl-β-d-glucopyranoside as a substrate and their potential for the breakdown of isoflavone glucosides to the biologically active aglycones in soymilk. Isoflavones quantification with HPLC and β-glucosidase activity were performed after 0, 12, 24, 36, and 48 h of incubation in soymilk at 37 °C. All six micro-organisms produced β-glucosidase, which hydrolysed the predominant isoflavone β-glucosides. There was a significant increase and decrease (P < 0.05) in the concentration of isoflavone aglycones and glucosides, respectively, in fermented soymilk. Based on the concentration of isoflavones during peak β-glucosidase activity, the hydrolytic potential was calculated. L. acidophilus 4461 had the highest aglycone concentration of 76.9% after 24 h of incubation, up from 8% in unfermented soymilk (at 0 h). It also had the best isoflavone hydrolytic index of 2.01, signifying its importance in altering the biological activity of soymilk.     

Bioactive action of β‐glucosidase enzyme of Bifidobacterium longum upon isoflavone glucosides present in soymilk

Bioconversion of isoflavone glucosides and antioxidant activity by probiotic strain (Bifidobacterium longum) during soymilk fermentation was investigated, as well as partial characterisation of the produced enzyme β‐glucosidase. The enzyme has higher affinity for genistin than for other substrates assayed. Maximum activity occurred at 42 °C and at pH 6.0; keeping 70–80% of activity for 60 days stored at low temperatures. Bifidobacterium longum grew well in soymilk (8.26 log CFU mL^?1 and pH of 3.9 at 24 h) and were produced in good quantities of organic acids. High hydrolysis degree of isoflavone glucosides (81.2%) was observed at 24 h. Enhancements in bioactivity were assessed in fermented soymilk by monitoring the radical‐scavenging activity, antioxidant activity and DNA protective action. The use of probiotic Bifidobacterium strain as β‐glucosidase producer increased bioactive isoflavone content and demonstrated that this enzyme plays a key role in the bioavailability of soymilk isoflavones, reducing the bioconversion time compared to other studies.     

植物乳杆菌58发酵豆乳产大豆异黄酮苷元条件的优化

本研究通过测定植物乳杆菌58在不同碳源中的生长和产β-葡萄糖苷酶情况,筛选菌株的最适碳源,并确定接种至豆乳的最佳时间.在接种量、糖含量、发酵时间和发酵温度等单因素实验基础上,根据Box-Behnken中心组合原理进行响应面实验设计,以大豆异黄酮苷元含量为指标,进一步优化菌株58发酵豆乳产大豆异黄酮苷元条件.结果显示,植...     

Soymilk phenolic compounds,isoflavones and antioxidant activity as affected by in vitro gastrointestinal digestion

The aim of this research was to evaluate changes in the phenolic compounds, isoflavones and antioxidant activity of soymilk following in vitro gastrointestinal digestion (including dialysis). Gastric digestion significantly influenced the release of bioactive substances from the soymilk matrix, increasing the concentration of total phenolic components (35% as the sum of individuals and 14% by Folin–Ciocalteu [F–C] method), total isoflavone content (22%) and total antioxidant activity (76%). The concentration of all those compounds was reduced significantly in the duodenal fraction in comparison to gastric digestion and their lowest concentration was observed in the dialysed fraction, where phenolic acids were not detected. The bioaccessibility of soymilk phenolic compounds was 15% as the sum of individuals and 20% by F–C assay; isoflavones 36% and constituents with antioxidant activity 27%. Results suggest that most of these compounds were sufficiently available to be absorbed and could contribute health benefits.     

A new biotechnological process to enhance the soymilk bioactivity

Equol, a daidzein metabolite produced exclusively by intestinal bacteria in some, but not all, humans, exhibits a wide range of beneficial health effects owing to its superior nutraceutical effect compared with isoflavones of soy. The aim of this work was to develop bioprocesses capable of increasing the bioactive properties of soymilk and, most importantly, increase the equol content by a biotechnological process in vitro. Biotransformation processes based on soymilk fermentation by probiotic lactic bacteria and application of the enzyme tannase caused an increase in the bioactive isoflavones and antioxidant capacity of soymilk. Furthermore, these processes approximately resulted in a 10-fold increase in the equol content of the soymilk. This is the first study to produce a significant equol concentration in soymilk using enzymatic processing only. The results suggest a new and effective biotechnological process, with major commercial potential, capable of producing a bioactive soy extract that intends to be “functional for everyone.”     

Changes in antioxidant capacity,isoflavone profile,phenolic and vitamin contents in soymilk during extended fermentation

In this study, the influence of lactic acid bacteria fermentation on the antioxidant capacity, phenolic compounds and vitamins profile of soymilk with different final fermentation pH values (pH 4.55, 4.15 and 3.85) was examined. Fermented soymilk (FSM) with a final pH of 3.85 exhibited relatively higher antioxidant capacity than samples terminated at pH 4.55 or 4.15, as shown from results of Folin–Ciocalteu assay (57.3–63.8 mg GAE/100 mL FSM), DPPH-radical scavenging efficiency (30.0–36.5%) and ferricyanide reducing power (1.13–1.58 mg AAE/100 mL FSM). Among all products, Lactobacillus rhamnosus WQ2-fermented soymilk showed the highest antioxidant level among the four Lactobacillus species, while Lactobacillus acidophilus CSCC 2400 possessed the best ability to deglycosylate isoflavone glucoside (IG), reducing IG concentration by 60.8%. Four vitamins, three phenolic acids, one flavanol, and six isoflavones were detected in fermented and non-fermented soymilk by HPLC-DAD. The contents of antioxidant compounds in FSM were closely related to reducing power and anti-radical ability. In addition, antioxidant capacity was highly correlated with proteolytic activity. This study also demonstrated the potential of extended fermentation to enhance the overall health-promoting property of the products.     

Hydrolysis of soybean isoflavone glycosides by a thermostable β-glucosidase from Paecilomycesthermophila

The β-glucosidase from Paecilomyces thermophila J18 was found to be capable of hydrolysing daidzin and genistin in a previous study. This report further evaluated the thermostability and hydrolysis of soybean isoflavone glycosides. The enzyme was found to be very stable at 50 °C, and retained more than 95% of its initial activity after 8 h at 50 °C. It converted isoflavone glycosides, in soybean flour extract and soybean embryo extract, to their aglycones, resulting in more than 93% of hydrolysis of three isoflavone glycosides (namely, daidzin, genistin and glycitin) after 4 h of incubation. Also, addition of the β-glucosidase greatly increased the contents of isoflavone aglycones in the suspended soybean flour and soymilk. The results indicate that the thermostable β-glucosidase may be used to increase the isoflavone aglycones in soy products. This is the first report on the potential application of fungal β-glucosidases for converting isoflavone glycosides to their aglycones in soy products.     

Sensory properties of soy yoghurts prepared from yellow and black soymilk using Streptococcus infantarius 12 and Weisellia sp. 4

BACKGROUND: Soy yoghurts containing enhanced bioactive isoflavone aglycones were prepared from combinations of yellow (YS) and black soymilk (BS) (YS:BS = 100:0, 70:30, 50:50 and 0:100, v/v) by fermentation with Streptococcus infantarius 12 and Weisellia sp. 4 at 37 °C for 10 h. Sensory properties of the bioactive soy yoghurts were investigated. Furthermore, other characteristics including the contents of isoflavones, viable cell numbers, pH, colour and DPPH radical scavenging activity were investigated. RESULTS: One hundred percent YS yoghurt showed the highest scores of all sensory attributes while 100% BS yoghurt was the lowest for colour and smoothness. The overall acceptability scores were 5.9 and 5.2 for 100% YS and 100% BS yoghurts, respectively (9‐point scale). The L‐values of soy yoghurts decreased as the percentage BS increased, indicating darkening of soy yoghurts. The contents of bioactive isoflavone aglycones (daidzein and genistein) in soy yoghurts remarkably increased (>1300 mg kg^?1) by fermentation. Viable cell numbers and pH of soy yoghurts ranged from 8.4 × 10⁸ to 9.6 × 10⁸ CFU g^?1 and 4.43 to 4.54, respectively. DPPH radical scavenging activity significantly increased as the percentage BS increased (P < 0.05). CONCLUSION: One hundred percent YS yoghurt was better for overall sensory properties while 100% BS yogurt was more bioactive in terms of the isoflavone aglycone contents and DPPH radical scavenging activity compared to other yoghurts. Copyright © 2008 Society of Chemical Industry     

Evaluation of the composition and concentration of isoflavones in soy based supplements,health products and infant formulas

Isoflavones are weak estrogenic compounds found in soybeans and soy foods. The objective of the present study was to determine the content and composition of isoflavones in different soy isoflavone supplements, soy based health products and infant formulas. Commercially available samples of 13 different soy supplements and 11 soy based health products obtained from South East Asia were analyzed for their isoflavone concentrations by using high-performance liquid chromatography and their peak identities confirmed by LC–MS. The extent of daily intake of isoflavones from different products based on the directions for their consumption was further evaluated. The amount of isoflavones in dietary supplements, soy based health products and infant formulas ranged from 405 to 57,570, 46.32–1333.80 and 59.54–226.84 μg/g of the sample, respectively, expressed as aglycone equivalents.     

Content and Profile of Isoflavones in Soy-Based Foods as a Function of the Production Process

Soy has been traditionally incorporated in diet as processed foods, such as soymilk, tofu, miso, tempeh, etc., and the consumption is commonly associated with a reduction of the development of chronic diseases due to their antioxidant, anti-inflammatory, and anti-allergic properties, among others. Many of the health benefits of soy have been attributed to isoflavones. They comprise a group of naturally occurring flavonoids consisting of heterocyclic phenols. Soy contains three types of isoflavones in four chemical forms: the aglycones daidzein, genistein, and glycitein; the β-glucosides daidzin, genistin, and glycitin; their 6″-O-malonyl-β-glucosides (6OMalGlc); and their 6″-O-acetyl-β-glucosides (6OAcGlc) conjugates. Industrial processing methods of soy-based food products commonly lead to the loss of isoflavones through removal of undesirable fractions. On the other hand, isoflavones can be transformed into different conjugates, which may have significant effects on the food texture and on the bioavailability and pharmacokinetics of the isoflavones. This article reviews the effect of a number of soybean processing treatments on the isoflavone content and profile. The preparation and manufacturing of different soy-based food and food ingredients, fermented and non-fermented, has been analyzed in terms of content and distribution of the three major isoflavone derivatives, daidzein, genistein, and glycitein, and their respective conjugates.     

Effect of free or protein-associated soy isoflavones on the antioxidant status in rats

BACKGROUND: The objective of this study was to investigate the effect of chronic ingestion of free and protein‐associated soy isoflavones on the antioxidant status in male Wistar rats. Free isoflavone (iso), protein‐associated soy isoflavone (iso + prot) and soy protein (prot) extracts were administered for 30 days by gavage to the rats at a dosage of 1 mg aglycone isoflavones per 200 g body weight, adjusted daily, and the prot group was given the same concentration of soy protein received by the iso + prot group. Antioxidant capacity of plasma, thiobarbituric acid‐reactive substance (TBARS) and glutathione (GSH) levels and catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities in plasma, erythrocytes and tissues and gene expression levels in liver and kidney were evaluated. RESULTS: Chronic ingestion of free but not of protein‐associated soy isoflavones nor of solely soy protein increased plasma antioxidant capacity and GPx activity in erythrocytes. Soy protein increased CAT activity and gene expression in liver. SOD activity in erythrocytes was increased by all treatments. CONCLUSION: The overall results confirm that dietary soy isoflavones have a positive effect on antioxidant status, enhancing antioxidant capacity of plasma and antioxidant enzymes in various tissues, but the effects are dependent on the form of administration and on a complex mechanism of antioxidant status balance on the organism. Copyright © 2010 Society of Chemical Industry     

Antioxidant capacity,phenolics and isoflavones in soybean by-products

This study aimed to determine proximate composition, antioxidant capacity, total phenolic content, isoflavones and free phenolic compounds in soy by-products. High carbohydrate and protein contents were found in grade A soymilk powder (GASP) compared to grade B soymilk powder (GBSP) and soy husk powder (SHP). Ash, moisture and total dietary fibre contents were reported to be the highest in soy husk, while GBSP had the highest fat content. Antioxidant capacity as assessed using β-carotene bleaching assay was in the order of SHP ≈ GBSP > GASP, and the ranking order of the Trolox Equivalent Antioxidant Capacity (TEAC) value was GASP ≈ GBSP > SHP, while the Ferric Reducing Antioxidant Power (FRAP) value was GASP > GBSP > SHP. The total phenolic content was in the range of 62.44–103.86 mg GAE/100 g wet weight, and the major phenolic compounds in free form were ferulic, vanilic as well as gallic acids. Acid hydrolysis increased the amount of total extractable isoflavone in all soy samples.     

Fermentation of calcium-fortified soymilk with Lactobacillus: effects on calcium solubility, isoflavone conversion, and production of organic acids

ABSTRACT:  The objective of this study was to enhance calcium solubility and bioavailability from calcium-fortified soymilk by fermentation with 7 strains of Lactobacillus , namely, L. acidophilus ATCC 4962, ATCC33200, ATCC 4356 , ATCC 4461 , L. casei ASCC 290, L. plantarum ASCC 276, and L. fermentum VRI-003. The parameters that were used are viability, pH, calcium solubility, organic acid, and biologically active isoflavone aglycone content. Calcium-fortified soymilk made from soy protein isolate was inoculated with these probiotic strains, incubated for 24 h at 37 °C, then stored for 14 d at 4 °C. Soluble calcium was measured using atomic absorption spectrophotometry (AA). Organic acids and bioactive isoflavone aglycones, including diadzein, genistein, and glycetein, were measured using HPLC. Viability of the strains in the fermented calcium-fortified soymilk was > 8.5 log₁₀ CFU/g after 24 h fermentation and this was maintained for 14-d storage at 4 °C. After 24 h, there was a significant increase ( P < 0.05) in soluble calcium. L. acidophilus ATCC 4962 and L. casei ASCC 290 demonstrated the highest increase with 89.3% and 87.0% soluble calcium after 24 h, respectively. The increase in calcium solubility observed was related to lowered pH associated with production of lactic and acetic acids. Fermentation significantly increased ( P < 0.05) the level of conversion of isoflavones into biologically active aglycones, including diadzein, genistein, and glycetein. Our results show that fermenting calcium-fortified soymilk with the selected probiotics can potentially enhance the calcium bioavailability of calcium-fortified soymilk due to increased calcium solubility and bioactive isoflavone aglycone enrichment.     

Critical Issues in R&D of Soy Isoflavone-enriched Foods and Dietary Supplements

ABSTRACT: Increasing consumer awareness of soy as a healthy food ingredient has led to a tremendous growth in sales of soy isoflavone‐enriched foods and dietary supplements in the past 3 y. Because of their weak estrogenic activity, isoflavones are believed to have preventive effects for several hormone‐dependent diseases. This article discusses some critical issues to be considered in any R&D program of novel soy and isoflavone products for the healthy‐food market, such as (1) Which isoflavone compounds should be in the product? (2) Which individual isoflavones are the most beneficial ones? (3) Can certain isoflavone groups be targeted to specific disease prevention goals? (4) Are there any safety concerns in isoflavone consumption? (5) How are isoflavones affected by thermal processing and storage? (6) How should isoflavones be analyzed? These questions may affect the choice of isoflavone source, processing conditions, quality control procedures, and marketing considerations.     

Stability of isoflavone phytoestrogens in fermented soymilk with Bifidobacterium animalis Bb12 during storage at different temperatures

Soy isoflavones in fermented soymilk with Bifidobacterium animalis Bb12 were stored at various temperatures (?80, 4, 24.8 and 37 °C) for 8 weeks and the concentration of isoflavones determined weekly using reverse‐phase high‐performance liquid chromatography (RP‐HPLC). The first‐order kinetic model was used to assess the degradation of each isomer at each storage temperature. Soymilk predominantly possesses high concentrations of isoflavone glucosides and very low concentration of bioactive aglycone component. During storage at various temperatures, concentrations of individual isoflavone isomers appeared to be significantly stable (P < 0.01). Interestingly, aglycones showed much smaller degradation constants compared with glucosides at all the storage temperatures studied. Genistein and daidzein were much more stable than glycitein and had almost similar degradation patterns, despite differences in their concentrations in the fermented soymilk. It was, however, observed that 4 °C was the most suitable storage temperature for the product in order to guarantee minimal degradation of bioactive isoflavone aglycones.     

Urinary isoflavone excretion in Korean adults: comparisons of fermented soybean paste and unfermented soy flour

BACKGROUND: This study compared the 24 h urinary excretion of isoflavone algycones after the consumption of fermented soybean paste or unfermented soy flour in a population of healthy Koreans (four males and five females). An equivalent amount (20 mg) of total isoflavone was consumed as either 52 g of soybean paste or 12.2 g of soy flour in a randomised, crossover trial consisting of two single‐time‐point feedings and having 3 day run‐in and washout periods between feedings. RESULTS: Analysis of starting materials indicated that daidzein (1.4%) and genistein (1.4%) constituted a smaller proportion of the total isoflavones in soy flour as compared with soybean paste (55.3% daidzein and 45.7% genistein). Free and glycosidic forms of isoflavones differed significantly between the two soy products. Levels of β‐glycosides as well as total glycosides (β‐glycosides, malonyl and acetyl forms) were two‐fold higher in soy flour as compared with soybean paste. Malonyl forms predominated in soy flour. CONCLUSIONS: The consumption of soybean paste resulted in significantly higher urinary excretion of both daidzein and genistein as compared with soy flour (P < 0.05 and P < 0.01 respectively). Copyright © 2007 Society of Chemical Industry