Exploring Impact of Probiotic-Fermented Soymilk on Dextran-Sulfate-Sodium-Induced Ulcerative Colitis via Modulating Inflammation and Gut Microbiota Profile.

SCOPE: Lactic acid bacteria with probiotic functions and their fermentation products play a role in regulating ulcerative colitis (UC). This study investigates the potential role of fermented soymilk (FSM4) rich in isoflavones on DSS-induced UC. METHODS AND RESULTS: Mice received 3% DSS and are supplemented daily once for 1 week by NFSM and FSM4. DSS usually causes intestinal inflammation and alters the gut microbiota. FSM4 intervention improves the UC-related inflammation and gut microbiota alteration. It considerably decreases pro-inflammatories such as TNF-α, IL-1ß, and IL-6 in serum and COX-2 and MPO in colon tissues and pathogenic bacteria (Escherichia-Shigella). This facilitates gut-healthy bacteria growth. These healthy bacteria negatively correlat with pro-inflammatory factors but positively associated with acetic acid, butyric acid, and propionic acid, which may act for PPAR-γ pathway activating and NF-κB p65 pathway inhibiting, lowering the risk of UC. Overall, FSM4 might alleviate UC and significantly reverse the dysbiosis of gut microbiota via the PPAR-γ activation. It could be a good alternative for developing functional food to protect against UC. CONCLUSION: FSM4 attenuates intestinal inflammation and modulates the SCFA-producing bacteria growth, which enable the PPAR-γ activation to alleviate the UC target, which could be a dietary intervention strategy for gut health.

The effects of soymilk plus probiotics supplementation on cardiovascular risk factors in patients with type 2 diabetes mellitus: a randomized clinical trial.

OBJECTIVE: Type 2 diabetes mellitus (T2DM) is associated with an increased risk of cardiovascular diseases. This study aimed to assess the effects of soymilk plus probiotics co-administration on cardiovascular risk factors in T2DM patients. METHODS: One hundred patients with T2DM (aged 40-75 years old) were randomly assigned into 4 groups (soymilk + probiotics supplement, soymilk + placebo, conventional milk + placebo, and probiotics supplement) for 6 weeks. Standard protocols were followed for the collection of fasting blood samples, dietary intakes, and anthropometric measurements. RESULTS: It was shown that soymilk + probiotics consumption significantly decreased diastolic blood pressure (DBP) (p = 0.001), triglycerides (TG) (P < 0.001), total cholesterol (TC) (p < 0.01), and insulin (P < 0.003) levels and significantly increased high-density lipoprotein cholesterol (HDL-C) (P = 0.002) levels. Soymilk + placebo administration significantly decreased DBP (p = 0.01), insulin (p = 0.006), and TG (p = 0.001) levels and significantly increased HDL-C (p = 0.03) levels. A significant decrease in insulin (p = 0.003) and systolic blood pressure (SBP) (p = 0.01) levels and an increase in HDL-C (p = 0.04) levels were observed after supplementation with probiotics. Findings from between-group comparisons showed a significant decrease in SBP levels in the probiotics supplement group compared to conventional milk group (p < 0.05). CONCLUSION: Soymilk and probiotics consumption might improve some cardiovascular risk factors in patients with T2DM. However, possible synergic effects while consumption of soymilk plus probiotics supplement didn't show in this study which warranted further research.

Riboflavin Bioenriched Soymilk Alleviates Oxidative Stress Mediated Liver Injury, Intestinal Inflammation, and Gut Microbiota Modification in B2 Depletion-Repletion Mice.

Epidemiological evidence emphasizes that ariboflavinosis can lead to oxidative stress, which in turn may mediate the initiation and progression of liver injury and intestinal inflammation. Although vitamin B2 has gained worldwide attention for its antioxidant defense, the relationship between B2 status, oxidative stress, inflammatory response, and intestinal homeostasis remains indistinct. Herein, we developed a B2 depletion-repletion BALB/c mice model to investigate the ameliorative effects of B2 bioenriched fermented soymilk (B2FS) on ariboflavinosis, accompanied by oxidative stress, inflammation, and gut microbiota modulation in response to B2 deficiency. In vivo results revealed that the phenotypic ariboflavinosis symptoms, growth rate, EGRAC status, and hepatic function reverted to normal after B2FS supplementation. B2FS significantly elevated CAT, SOD, T-AOC, and compromised MDA levels in the serum, simultaneously up-regulated Nrf2, CAT, and SOD2, and down-regulated Keap1 gene in the colon. The histopathological characteristics revealed significant alleviation in the liver and intestinal inflammation, confirmed by the downregulation of inflammatory (IL-1ß and IL-6) and nuclear transcription (NF-κB) factors after B2FS supplementation. B2FS also increased the abundance and diversity of gut microbiota, increased the relative abundance of Prevotella and Absiella, as well as decreased Proteobacteria, Fusobacteria, Synergistetes, and Cyanobacteria in strong conjunction with antioxidant, anti-inflammatory properties, and gut homeostasis along with the remarkable increase in cecal SCFAs content. We hereby reveal that B2FS can effectively alleviate deleterious ariboflavinosis associated with oxidative stress mediated liver injury, chronic intestinal inflammation, and gut dysbiosis in the B2 depletion-repletion mice model via activation of the Nrf2 signaling pathway.

Functional relevance and health benefits of soymilk fermented by lactic acid bacteria.

The growing interest of consumers towards nutritionally enriched, and health promoting foods, provoke interest in the eventual development of fermented functional foods. Soymilk is a growing trend that can serve as a low-cost non-dairy alternative with improved functional and nutritional properties. Soymilk acts as a good nutrition media for the growth and proliferation of the micro-organism as well as for their bioactivities. The bioactive compounds produced by fermentation of soymilk with lactic acid bacteria (LAB) exhibit enhanced nutritional values, and several improved health benefits including antihypertensive, antioxidant, antidiabetic, anticancer and hypocholesterolaemic effects. The fermented soymilk is acquiring a significant position in the functional food industry due to its increased techno-functional qualities as well as ensuring the survivability of probiotic bacteria producing diverse metabolites. This review covers the important benefits conferred by the consumption of soymilk fermented by LAB producing bioactive compounds. It provides a holistic approach to obtain existing knowledge on the biofunctional attributes of fermented soymilk, with a focus on the functionality of soymilk fermented by LAB.

Effect of freezing treatment of soybean on soymilk nutritional components, protein digestibility, and functional components.

Soymilk is a popular beverage in many countries owing to its nutrition and health effects. To increase household consumption of soymilk, instant soybeans were developed by freezing and subsequent drying pretreatment, which overcome the time-consuming need of soaking during soymilk preparation for home making. However, compared with the traditional soymilk making, the nutritional quality and functional properties of this soymilk made from the soybean by direct grinding in water without soaking are not clear yet. Soymilk made from untreated soybeans, soaked soybeans, and soaking, freezing, and air-drying soybeans (FADTS) were compared on their properties including nutritional components, in vitro protein digestibility, and functional components. It was found that FADTS was the best at extracting lipid and Ca, good at extracting of protein, carbohydrate, oligosaccharides, Fe, phytic acids, and tannins, and in producing soymilks with highest in vitro protein digestibility. The soluble protein and protein digestibility of FADTS (4 day) increased significantly from 44.4% and 78.5% of control to 56.2% and 85.0%, respectively. Soymilk from 4 days FADTS contained similar protein content and higher Fe content (4.40 mg/kg) compared to soaked sample (3.82 mg/kg). The results revealed that FADTS performed better at producing soymilk than untreated and soaked soybeans.

Effect of Fermented Soymilk-Honey from Different Probiotics on Osteocalcin Level in Menopausal Women.

Osteoporosis has been discovered to be a risk factor for menopausal women. Although synbiotics (probiotics and prebiotics) are found in fermented soymilk-honey made using local probiotics, their effect on osteocalcin levels is still unknown. Therefore, this study's objective was to determine the influence of fermented soymilk-honey from different probiotics on osteocalcin levels. A 90-day pre-post quasi-experimental study with a control design was conducted on 54 postmenopausal women divided into three intervention groups namely, the soymilk (SM) group, the soymilk-honey fermented with Lactobacillus casei subsp. casei R-68 (SMH Lc) group, and the soymilk-honey fermented with Lactobacillus plantarum 1 R 1.3.2 (SMH Lp) group. Participants consumed 100 mL of soymilk (SM) or fermented soymilk with honey (SMH Lc or SMH Lp) for 90 days. At the beginning and end of the study, the blood serum osteocalcin level was measured and subjects' health status was assessed, such as cholesterol total, random blood glucose, and uric acid levels. Our results presented that in the SMH Lp group, 90 days supplementation of soy-honey milk fermented with Lactobacillus plantarum 1 R 1.3.2 significantly reduced the level of blood serum osteocalcin. Based on these results it is justified to perform more detailed studies on the effect of fermented soy-honey milk on bone health.

Pepsin Diffusivity and In Vitro Gastric Digestion of Soymilk as Affected by Binding of Tea Polyphenols to Soy Proteins.

This study aimed to investigate the effect of tea polyphenol extract (TPE) on the in vitro gastric digestion of soymilk. Fluorescence recovery after photobleaching was applied to measure pepsin diffusivity in soymilk. The characteristics of soymilk digesta were evaluated by gel electrophoresis, degree of hydrolysis (DH), molecular weight distribution, free amino acid analysis, particle size, antioxidant capacity, and trypsin/chymotrypsin inhibitor activity (TIA/CIA). The binding between soy proteins and tea polyphenols could significantly impair in vitro gastric digestion of soymilk by decreasing pepsin diffusivity from 91.3 to 70.3 µm2/s and DH from 17.13 to 13.93% with 1.2 mg/g TPE addition. Soymilk with 0.6 mg/g TPE addition exhibited low TIA/CIA and a strong antioxidant capacity in gastric digesta, which might be good for the following intestinal digestion. A better understanding of the effect of polyphenol on the digestion of protein-based food may be beneficial to innovation in food manufacturing.

Glutamate Decarboxylase (GAD) Extracted from Germinated Rice: Enzymatic Properties and Its Application in Soymilk.

Glutamate decarboxylase (GAD) is an important enzyme in biological metabolisms acting on catalyzing the irreversible α-decarboxylation of L-glutamic acid to γ-aminobutyric acid (GABA) and CO2, which was focused in this study. Three rice varieties different in color were germinated at different times and used for crude GAD extraction. Crude GADs with an optimal germination time from germinated black (GBR), red (GRR), and white (GWR) rice were evaluated for enzymatic properties, including the effect of pHs, temperatures, and concentrations of both L-glutamic acid and pyridoxal 5'-phosphate (PLP). Crude GAD with optimum enzymatic properties was selected to be partially purified using ammonium sulfate (AMS) precipitation. The obtained GAD was supplemented to soymilk and determined for GABA content. All crude GADs from germinated rice at 10 germination days presented the highest enzyme activity. For enzymatic properties, crude GADs showed the highest activity at pH in a range of 5.6-6.0 at 60ºC. The Km values of crude GADs were in the range of 7.68-8.06 mM for L-glutamic acid and 0.15-0.20 µM for PLP and were the lowest in crude GAD from GBR. GAD from GBR presented the highest enzyme activity in the fraction with 50% saturation (v/v) after AMS precipitation and it was purified for 14.61 folds. The addition of this GAD (1.0%, v/v) resulted in the increasing of GABA content in soymilk to 53.79 mg/100 mL, accounted for 1.23 times compared with control.

Co-Ingestion of Rice Bran Soymilk or Plain Soymilk with White Bread: Effects on the Glycemic and Insulinemic Response.

The regular consumption of soy products is associated with inverse incidence of type 2 diabetes, and there has been an increasing interest in the glycemia reducing potential of rice bran and its components. In this study, we investigated whether consuming soymilk with the addition of rice bran (fiber) can reduce the glycemic response of a carbohydrate meal. Seventeen healthy Asian men (BMI: 18.5-29 kg/m²) participated in this randomized crossover trial. On four occasions, they consumed white bread (two times) and white bread with two different soymilks differing in protein and rice bran content. Blood samples were taken to measure glucose and insulin response over a period of 3 hours. Taking the glycemic index (GI) value of white bread as a reference value of 100, the GI of white bread when co-ingested with rice bran soymilk (RBS) was 83.1 (±7.7) and sugar-free soymilk (SFS) was 77.5 (±10.1), both were lower than white bread (p < 0.05). The insulin response of both soymilk treatments was similar to white bread (p > 0.05). The glucose/insulin ratio of RBS and SFS were respectively 43.1 (± 6.1) and 60.0 (± 17.0) and were lower (p < 0.05) than white bread (123.5 ± 21.1) during the first 30 min. In conclusion, co-ingestion of low amounts of soy protein with a carbohydrate meal stimulated early-phase insulin secretion and thereby increased blood glucose clearance effectiveness. Furthermore, rice bran-fortified soymilk reduced the glycemic response similarly to soymilk with a greater dose of soy protein. Rice bran and its components offer therapeutic potential for glycemic and insulinemic control.

Consumption of Soymilk Reduces Lipid Peroxidation But May Lower Micronutrient Status in Apparently Healthy Individuals.

Consumption of soy products is speculated to reduce the risk and progression of some disease conditions. The underlying mechanisms mediating this effect are uncertain, but the lowering of oxidative stress has been suggested. This study was aimed at investigating the effect of soymilk on antioxidant status and lipid peroxidation in apparently healthy individuals. Five hundred milliliters of soymilk was taken daily by each of 39 apparently healthy individuals for a period of 28 days. Two sets of blood samples (baseline and after 28 days of soymilk intake) were collected and assayed for total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione S-transferase (GST), manganese (Mn) and zinc (Zn) levels, using standard methods. Consumption of soymilk significantly increased serum TAC and reduced serum MDA when compared with baseline values (P < .001, P < .001 respectively). The decrease in MDA concentration was significantly contributed by the women (P < .001). Mean serum Mn was also significantly reduced (P = .03) when compared with baseline values. There were no changes in serum Zn concentration and the activity of SOD enzyme. The serum GST activity was significantly increased in men (P = .02) and significantly reduced in women (P < .001) in comparison with their corresponding baseline values. Daily consumption of soymilk enhanced antioxidant status and this led to reduced lipid peroxidation. It also resulted in a significant reduction of Mn. The dietary use of soymilk as an adjuvant to supplement meals seems beneficial health wise. However, soymilk should be taken with caution as it could result in micronutrient deficiency.

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model.

Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free fatty acid-induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with palmitic acid (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free fatty acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.

Effect of Fermentation pH on Protein Bioaccessibility of Soymilk Curd with Added Tea Polyphenols As Assessed by in Vitro Gastrointestinal Digestion.

The aim of this study was to compare the effect of fermentation pH on protein bioaccessibility of four soymilk curds enriched with tea polyphenols (TP). The curds were generated by fermentation with Weissella hellenica D1501 and the fermentation terminated at different pH values, namely at pH 5.7, 5.4, 5.1, and 4.8 (SMTP-5.7, SMTP-5.4, SMTP-5.1, SMTP-4.8). Particle-size distribution, soluble protein content, gel electrophoresis, and peptides content were monitored at oral, gastric, and intestinal levels. Results showed that SMTP-4.8 was the matrix most resistant to protein digestion in the gastric phase according to the soluble protein content. Similar particle size distribution and protein degradation patterns were observed for these curds in gastric and intestinal phase. However, there was a significant difference (P < 0.05) in the content of small peptides (<10 kDa) at the end of intestinal digestion among the four curds. Overall, terminating fermentation at pH 5.4-5.7 of soymilk curds enriched with TP is recommended.

Passion fruit by-product and fructooligosaccharides stimulate the growth and folate production by starter and probiotic cultures in fermented soymilk.

Two starter cultures (Streptococcus (St.) thermophilus ST-M6 and TA-40) and five probiotic strains (St. thermophilus TH-4, Lactobacillus (Lb.) acidophilus LA-5, Lb. rhamnosus LGG, Lb. fermentum PCC, and Lb. reuteri RC-14) were used to ferment different soymilk formulations supplemented with passion fruit by-product and/or fructo-oligosaccharides (FOS) with the aim of increasing folate concentrations. Growth and folate production of individual strains were evaluated and the results used to select co-cultures. Both St. thermophilus ST-M6 and TH-4 were the best folate producers and were able to increase the folate content of all soymilk formulations when used alone or in co-culture with lactobacilli strains, especially in the presence of both passion fruit by-product and FOS. Thus, passion fruit by-product and FOS could be used as dietary ingredients to stimulate the folate production by selected bacterial strains during the fermentation of soymilk. It was also shown that vitamin production by microorganisms is strain-dependent and may also be influenced by nutritional and environmental conditions.

Soymilk enriched with green coffee phenolics - Antioxidant and nutritional properties in the light of phenolics-food matrix interactions.

This study investigated the effect of soymilk fortification with green coffee extract (GCE) on phenolic contents, antioxidant capacity, relative in vitro digestibility of proteins and starch, and consumer acceptance. Special attention was paid to the effect of phenolics-food matrix interactions on fortification efficiency. Soymilk was enriched with GCE extracts containing 0.025-1mg of phenolics per 1mL-samples M1-M6. Compared to control, an increase in phenolic contents of up to 70% (M6) was observed for potentially bioaccessible fractions (AD). The antiradical activity and reducing power were also about 1.9 and 10.1 times higher, respectively. However, the determined phenolic and antioxidant activities differed from those predicted. Fortification improved the digestibility of nutrients when higher doses of GCE was introduced (M4-M6). The addition of GCE at an adequate dose allowed the production of a beverage with elevated hedonic properties. In conclusion, fortification was a successful in improving the pro-health status of soymilk.

Innovative Soaking and Grinding Methods and Cooking Affect the Retention of Isoflavones, Antioxidant and Antiproliferative Properties in Soymilk Prepared from Black Soybean.

This study's objective was to characterize the effect of traditional and 3 newly devised (soaking+grinding) methods combined with cooking on the content and composition of phenolic substances, antioxidant, and antiproliferative properties of soymilk prepared from black soybean. Phenolic substances and antioxidant profile were characterized and antiproliferation of prostate cancer DU145 cells was conducted using a cell culture assay. Results indicated Grinding Method 4 produced significantly (P < 0.05) higher total phenolic content (TPC), total flavonoid content (TFC), condensed tannin content (CTC), and total isoflavone content in both raw and cooked black soymilk as compared to Method 1. Cooking soymilk reduced 23% to 38% of total phenolic substances. Raw black soymilk produced by Method 4 displayed the highest antioxidant capability, which was determined using ORAC, FRAP, and DPPH assays, and a higher antiprostate cell proliferation ability. Cooking only slightly reduced the potency to inhibit DU145 prostate cancer cells as IC50 value was increased from the average of about 4.0 mg/mL of raw soymilk extracts to 5.5 mg/mL of cooked soymilk extracts of all grinding methods. Overall, total isoflavone content was the only component that was negatively correlated with IC50 value (r = -0.93, P < 0.05) which indicates the ability to inhibit prostate cancer cell is associated with the increase in total isoflavone content, not with any other phenolic substances or antioxidant properties.

Latifolicinin A from a Fermented Soymilk Product and the Structure-Activity Relationship of Synthetic Analogues as Inhibitors of Breast Cancer Cell Growth.

The functional components in soymilk may vary depending upon the fermentation process. A fermented soymilk product (FSP) obtained by incubation with the microorganisms of intestinal microflora was found to reduce the risk of breast cancer. Guided by the inhibitory activities against breast cancer cells, two cytotoxic compounds, daidzein and (S)-latifolicinin A, were isolated from the FSP by repetitive extraction and chromatography. Latifolicinin A is the n-butyl ester of ß-(4-hydroxyphenyl)lactic acid (HPLA). A series of the ester and amide derivatives of (S)-HPLA and L-tyrosine were synthesized for evaluation of their cytotoxic activities. In comparison, (S)-HPLA derivatives exhibited equal or superior inhibitory activities to their L-tyrosine counterparts, and (S)-HPLA amides showed better cytotoxic activities than their corresponding esters. In particular, (S)-HPLA farnesyl amide was active to triple-negative MDA-MB-231 breast cancer cells (IC50 = 27 µM) and 10-fold less toxic to Detroit-551 normal cells.

Development of an efficient soymilk cream production method by papain digestion, heat treatment, and low-speed centrifugation.

We developed the simple method of soymilk cream production from the high-fat soymilk, which was prepared by papain digestion and heat treatment. As a result of the treatment, high-fat soymilk was aggregated and it became possible to separate soymilk cream as the surface fraction by low-speed centrifugation (6000 × g, 10 min).

Influence of tea extract supplementation on bifidobacteria during soymilk fermentation.

In this study, the influence of tea extract (TE) supplementation on the viability and membrane lipid compositions of Bifidobacterium was investigated. Fermented soymilk-tea (SMT) was produced by culturing selected bifidobacteria in soymilk supplemented with green or black TE. Culturability of four bacteria in the presence of various concentrations of TE was examined by plate count method. Bifidobacterium longum CSCC 5089 (BL5089) and B. longum CSCC 5022 (BL5022) were selected for further study based on their sensitivity to TE. The effect of TE supplementation on bacterial cell viability and integrity was assessed by flow cytometry in combination with fluorescence probes. Total lipids of bacterial cell were extracted using an enzyme-assistant extraction method. Fatty acids (FAs) were determined and quantified by GC-MS. Phospholipids (PLs) were separated by high performance thin-layer chromatography (HPTLC) and their relative abundances were determined by densitometry. Total tea phenolic content (TTP) in SMTs with varying concentrations of TE was quantified by HPLC. Among the four Bifidobacterium monitored, TE only significantly inhibited BL5089 (p<0.01) in a dose-dependent manner, with minimum inhibition concentrations (MICs) determined to be 15.45mg/mL TTP for green TE and 7.34mg/mL TTP for black TE. Flow cytometric analysis revealed different staining patterns of cell populations and compromise in cell integrity upon exposure to high concentrations of TE. Results from GC-MS showed that unsaturated to saturated FA ratios significantly decreased (p<0.01) in the membrane of BL5089 cells upon TE exposure. Separation of PLs by HPTLC showed dramatic alterations in phosphatidylcholine and phosphatidylglycerol contents due to TE treatment.

Antiradical and tea polyphenol-stabilizing ability of functional fermented soymilk-tea beverage.

This study examined the potential of two-step fermentation to preserve TPs in functional soy-tea beverage. Fermented soymilk-tea (FST) was produced by culturing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus and Bifidobacterium longum in soymilk supplemented with tea extract (TE). Total phenolic content (TPC) and anti-radical activities were determined for FSTs and fermented soymilk (FS). A HPLC method was employed to quantify nine major tea phenolics in FST products. TPC was significantly higher (p<0.05) in FST than FS, in the order of green tea>oolong tea>black tea>soymilk. The FSTs were effective at scavenging DPPH-radical rather than hydroxyl radical. Optimal pH to stabilize TPs in SMT was ca. 5.7, which reduced total TP loss by ca. 40% compared with that obtained from products with TE supplemented at the beginning of fermentation. A gradual decrease in TPs was observed during storage (4 °C), with more than half of total TPs remained in FST after 8 weeks.

Quantitative determination of active Bowman-Birk isoinhibitors, IBB1 and IBBD2, in commercial soymilks.

Naturally-occurring serine protease inhibitors of the Bowman-Birk family exert their potential chemopreventive and/or therapeutic properties via protease inhibition. In this study, we have quantified the amounts of active BBI isoinhibitors, IBB1 and IBBD2, in six commercial soymilks. By using cation exchange chromatography, the BBI isoinhibitors were isolated and their specific trypsin inhibitory activity was used to estimate their amounts in soymilk samples. IBB1 and IBBD2 concentrations ranged from 0.44 to 5.20 and 0.27 to 4.60 mg/100ml of soymilk, respectively; total BBI, considered as the sum of both isoinhibitors, ranged from 0.60 to 9.07 mg/100ml of soymilk. These data show that physiologically relevant amounts of active BBI are present in commercial soymilk and may exert potential health-promoting effects.