Blue Whiting (Micromesistius poutassou) Protein Hydrolysates Increase GLP-1 Secretion and Proglucagon Production in STC-1 Cells Whilst Maintaining Caco-2/HT29-MTX Co-Culture Integrity.

Inducing the feeling of fullness via the regulation of satiety hormones presents an effective method for reducing excess energy intake and, in turn, preventing the development of obesity. In this study, the ability of blue whiting soluble protein hydrolysates (BWSPHs) and simulated gastrointestinal digested (SGID) BWSPHs, to modulate the secretion and/or production of satiety hormones, such as glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK) and peptide YY (PYY), was assessed in murine enteroendocrine STC-1 cells. All BWSPHs (BW-SPH-A to BW-SPH-F) (1.0% w/v dw) increased active GLP-1 secretion and proglucagon production in STC-1 cells compared to the basal control (Krebs-Ringer buffer) (p < 0.05). The signaling pathway activated for GLP-1 secretion was also assessed. A significant increase in intracellular calcium levels was observed after incubation with all BWSPHs (p < 0.05) compared with the control, although none of the BWSPHs altered intracellular cyclic adenosine monophosphate (cAMP) concentrations. The secretagogue effect of the leading hydrolysate was diminished after SGID. Neither pre- nor post-SGID hydrolysates affected epithelial barrier integrity or stimulated interleukin (IL)-6 secretion in differentiated Caco-2/HT-29MTX co-cultured cells. These results suggest a role for BWSPH-derived peptides in satiety activity; however, these peptides may need to be protected by some means to avoid loss of activity during gastrointestinal transit.

Antifungal activity of a de novo synthetic peptide and derivatives against fungal food contaminants.

The development of novel solutions to fight microbial food contaminants rests upon two pillars, which are the development of resistant strains and consumers' desire for a reduced consumption of synthetic drugs. Natural antimicrobial peptides possess the qualities to overcome these issues. De novo synthesis of novel antifungal compounds is a major progress that has been facilitated by the identification of parameters involved in the antimicrobial activity. A 14-residue peptide named KK14, with the sequence KKFFRAWWAPRFLK-NH2 , was designed and inhibited conidial germination and fungal growth of food contaminants within the range 6.25 to 50 µg/ml and 6.25 to 100 µg/ml, respectively. The study of three analogues of the peptide highlighted the role of some residues in the structural conformation of the peptide and its antifungal activity. The substitution of a Pro residue with Arg increased the helical content of the peptide not only its antifungal activity but also its cytotoxicity. The insertion of an unnatural bulky residue ß-diphenylalanine or a full d-enantiomerization overall increased the antifungal potency. The four peptides showed similar behaviour towards salt increase, heat treatment, and pH decrease. Interestingly, the denantiomer remained the most active at high pH and after proteolytic digestion. The four peptides did not present haemolytic activity up to 200 µg/ml but had different behaviours of cytotoxicity. These differences could be crucial for potential application as pharmaceutical or food preservatives.

Cereal type significantly affects the composition and reconstitution characteristics of dried fermented milk-cereal composites.

BACKGROUND: Dairy and cereal products are frequently combined to create composites with enhanced nutritional benefits. Commercially available dried dairy-cereal composites are typically reconstituted and cooked to produce porridge or soup. RESULTS: Dried fermented milk-cereal composites (FMCC) with ∼193 g kg-1 protein were prepared by blending fermented milk with parboiled oats (FMCCo), wheat (FMCCw), or barley (FMCCb), incubating the blend, drying, and milling. Cereal type significantly affected the composition of the FMCC and the properties of the reconstituted, cooked FMCC (R-FMCC). The FMCCo had a higher starch and fat content and lower levels of lactose, lactic acid, and amylose than FMCCb. The R-FMCCo had higher viscosity during cooking at 95 °C and cooling to 35 °C, and higher values of yield stress (σ0 ), consistency index (K) and viscosity on shearing from 20 to 120 s-1 at 60 °C than R-FMCCb. The FMCCw had lower levels of fat and ß-glucan than FMCCo or FMCCb, but was otherwise closer to FMMCb with respect to composition, cooking properties and flow behavior. CONCLUSION: Differences in composition and consistency associated with cereal type are likely to affect the nutritional value of the FMCC. © 2018 Society of Chemical Industry.

Concurrent iron and zinc deficiencies in lactating mothers and their children 6-23 months of age in two agro-ecological zones of rural Ethiopia.

PURPOSE: The aim of this study is to examine the co-occurrences of low serum ferritin and zinc and anaemia among mothers and their children in two agro-ecological zones of rural Ethiopia. METHODS: Data were collected from 162 lactating mothers and their breast fed children aged 6-23 months. The data were collected via a structured interview, anthropometric measurements, and blood tests for zinc, ferritin and anaemia. Correlation, Chi-square and multivariable analysis were used to determine the association between nutritional status of mothers and children, and agro-ecological zones. RESULTS: Low serum levels of iron and zinc, anaemia and iron deficiency anaemia were found in 44.4, 72.2, 52.5 and 29.6% of children and 19.8, 67.3, 21.8, 10.5% of mothers, respectively. There was a strong correlation between the micronutrient status of the mothers and the children for ferritin, zinc and anaemia (p < 0.005). Deficiency in both zinc and ferritin and one of the two was observed in 19.1, and 53.7% of the mothers and 32.7 and 46.3%, of their children, respectively. In the 24 h before the survey, 82.1% of mothers and 91.9% of their infants consumed foods that can decrease zinc bioavailability while only 2.5% of mothers and 3.7% of their infants consumed flesh foods. CONCLUSION: This study shows that micronutrient deficiencies were prevalent among lactating mothers and their children, with variation in prevalence across the agro-ecological zones. This finding calls for a need to design effective preventive public health nutrition programs to address both the mothers' and their children's needs.

Levels of potential bioactive compounds including carotenoids, vitamin C and phenolic compounds, and expression of their cognate biosynthetic genes vary significantly in different varieties of potato (Solanum tuberosum L.) grown under uniform cultural conditions.

BACKGROUND: In addition to their high carbohydrate content, potatoes are also an important dietary source of vitamin C and bioactive secondary metabolites, including phenolic compounds and carotenoids, which have been suggested to play a role in human health. The expression of genes encoding key enzymes involved in the synthesis of these compounds was assessed by reverse transcription-quantitative polymerase chain reaction and compared to the accumulation of the corresponding product in seven potato varieties showing contrasting levels of metabolite accumulation. RESULTS: Strong positive correlations were found between phenolic content in the flesh of tubers and transcript levels of phenylalanine ammonia lyase (PAL) and chalcone synthase (CHS) genes. The expression of PAL and CHS was also related to that of AN1, a transcription factor involved in the synthesis of anthocyanins, suggesting that these genes are regulated in a coordinated manner. No clear relationship was found between transcript levels of phytoene synthase (PSY) or L-galactono-1,4-lactone dehydrogenase (GLDH) genes and total carotenoid or vitamin C accumulation, respectively. CONCLUSION: Data indicate that levels of total phenolic and flavonoid compounds in potato are controlled primarily by PAL and CHS gene expression. Transcript levels of PSY and GLDH did not control accumulation of carotenoids or vitamin C.

Antioxidant activities and selected characteristics of gelatin hydrolysates from seabass (Lates calcarifer) skin as affected by production processes.

Antioxidant activities and selected characteristics of gelatin hydrolysates from seabass skin as affected by production processes were investigated. Hydrolysates were prepared using different processes, including hydrolysis during and after gelatin extraction. Samples hydrolysed during gelatin extraction showed a higher degree of hydrolysis (DH) and yield compared with those hydrolysed after gelatin extraction (p < 0.05). All hydrolysates had a creamy yellowish colour. A lower abundance of volatile compounds was found in the hydrolysates produced during gelatin extraction, in comparison with those obtained after gelatin extraction. Hydrolysates prepared during gelatin extraction had higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidative power (FRAP) and ferrous ion chelating activity (p < 0.05). Following a simulated in vitro gastrointestinal digestion, the DPPH radical scavenging activity and FRAP of the hydrolysates was retained, whilst ferrous ion chelating activity increased. The most appropriate conditions for the generation of antioxidant hydrolysates from seabass skin were identified.

Immunomodulatory potential of a brewers' spent grain protein hydrolysate incorporated into low-fat milk following in vitro gastrointestinal digestion.

Brewers' spent grain (BSG) protein rich fraction was previously hydrolysed using Alcalase (U) and three additional fractions were prepared by membrane fractionation; a 5-kDa retentate (U > 5), a 5-kDa permeate (U < 5) and a 3-kDa permeate (U < 3). In the present study, these fractions were added to milk, subjected to simulated gastrointestinal digestion (SGID) and their anti-inflammatory potential was investigated. The digestates caused a significant reduction (p < 0.05) in interleukin-6 (IL-6) production in Concanavalin-A (ConA)-stimulated Jurkat T cells. The samples did not significantly alter the production of IL-6 in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. IL-2 and interferon-γ (IFN-γ) production in stimulated Jurkat T cells and IL-1ß and tumor necrosis factor-α (TNF-α) production in stimulated RAW 264.7 cells were not affected in the presence of the digestates. Results show that a SGID milk product supplemented with BSG hydrolysate and its associated ultrafiltered fractions can confer anti-inflammatory effects in Jurkat T cells.

A study of the ability of bioactive extracts from brewers' spent grain to enhance the antioxidant and immunomodulatory potential of food formulations following in vitro digestion.

Bioactivity of a snack-bar, chocolate-drink and yogurt fortified with brewers' spent grain (BSG) phenolic extracts (P2 or B2) or protein hydrolysates (barley protein hydrolysate (BPH), BPH < 3 kDa, BPH < 5 kDa, BPH > 5 kDa) was measured following gastrointestinal in vitro digestion. Concentrations of 0.5 and 0.1% (v/v) digestates were chosen for addition to Caco-2 and Jurkat T cells, respectively. Yogurt and B2 digestate protected against H2O2-induced DNA damage in Caco-2 cells (p < 0.05), by the comet assay. Snack-bar digestates possessed significant (p < 0.05) immunomodulatory effects, measured by ELISA in concanavalin-A stimulated Jurkat T cells. Addition of BPH enhanced (p < 0.05) the IFN-γ reducing capacity of the snack-bar while addition of BPH < 3 and < 5 kDa reduced IL-2 production to a greater extent than unfortified yogurt (p < 0.05). Selected BSG components can enhance the antioxidant and anti-inflammatory potential of foods.

Recent advances in Phytosterol Oxidation Products.

Phytosterols and their oxidation products have become increasingly investigated in recent years with respect to their roles in diet and nutrition. We present a comprehensive review of recent literature on Phytosterol Oxidation Products (POP) identifying critical areas for future investigation. It is evident that POP are formed on food storage/preparation; are absorbed and found in human serum; do not directly affect cholesterol absorption; have evidence of atherogenicity and inflammation; have distinct levels of cytotoxicity; are implicated with high levels of oxidative stress, glutathione depletion, mitochondrial dysfunction and elevated caspase activity.

Phenolic-enriched fractions from brewers' spent grain possess cellular antioxidant and immunomodulatory effects in cell culture model systems.

BACKGROUND: Large quantities of brewers' spent grain (BSG), a co-product of the brewing industry, are produced annually. BSG contains hydroxycinnamic acids, and phenolic-rich extracts from BSG have previously demonstrated the ability to protect against oxidant-induced DNA damage. The present study investigated the anti-inflammatory potential of eight phenolic extracts from BSG: four pale (P1-P4) and four black (B1-B4) extracts. RESULTS: BSG extracts were more cytotoxic in Jurkat T than U937 cells, with lower IC50 values in Jurkat T cells, measured using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Pale BSG extracts P2 and P3 showed the greatest anti-inflammatory potential, significantly (P < 0.05) reducing interleukin-2 (IL-2), interleukin-4 (IL-4, P2 only), interleukin-10 (IL-10) and interferon-γ (IFN-γ) production. In addition, extracts P1-P3 and B2-B4 showed significant (P < 0.05) antioxidant effects, determined by the cellular antioxidant activity assays superoxide dismutase, catalase and glutathione content (GSH). CONCLUSION: Phenolic extracts from BSG, particularly the pale BSG extracts, have the ability to reduce a stimulated cytokine production and may also protect against cellular oxidative stress. Results of the present study highlight the potential of BSG phenolic extracts to act as functional food ingredients, providing an alternative use and improving the value of this brewing industry co-product.

High-Pressure Processing on Whole and Peeled Potatoes: Influence on Polyphenol Oxidase, Antioxidants, and Glycaemic Indices.

Polyphenol oxidase (PPO) inactivation in five whole and peeled Irish potato cultivars was investigated using high-pressure processing (HPP) at 400 MPa and 600 MPa for 3 min. PPO activity was significantly lower in most of the HPP-treated samples, while the highest PPO inactivation was observed after HPP at 600 MPa. No significant (p > 0.05) changes were observed on the total phenolic content and antioxidant activity of all the HPP-treated potatoes. Regarding individual phenolic acids, chlorogenic acid was decreased significantly (p < 0.05) in all studied varieties with a concomitant increase (p < 0.05) in caffeic and quinic acid. Similarly, ferulic acid was also increased (p < 0.05) in all studied varieties after the HPP treatment, while there was a variation in rutin and 4-coumaric acid levels depending on the cultivar and the sample type. Anthocyanins in the coloured whole potato varieties (i.e., Kerr's Pink and Rooster), tentatively identified as pelargonidin-O-ferulorylrutinoside-O-hexoside and pelargonidin-O-rutinoside-O-hexoside, also exhibited significantly (p < 0.05) higher levels in the HPP-treated samples as opposed to those untreated. Glycaemic indices of the potatoes treated with HPP did not differ with the corresponding untreated cultivars.

The impact of thermal processing on the simulated infant gastrointestinal digestion, bactericidal and anti-inflammatory activity of bovine lactoferrin - An in vitro study.

Lactoferrin (LF) is a multifunctional glycoprotein which, when thermally processed, undergoes significant physicochemical changes. The link between such changes and the bioactivity of LF is not well characterised and requires much research. In this work, bovine LF solutions (1%, w/v, protein, pH 7) were thermally processed using high temperature short time conditions (72, 80, 85 or 95 °C with 15 s holding times). Following this, it was shown that LF and heat induced LF aggregates were largely resistant to simulated infant gastric, but not intestinal, digestion. Also, the efficacy of LF bactericidal activity, and inhibition of lipopolysaccharide-induced NF-κB activation were negatively impacted by thermal processing. This study confirmed that the efficacy of LF bio-functionalities was affected by the extent of heat-induced changes in protein structure whereby processing conditions of least severity (i.e. pasteurisation) had the least impact on bioactivity.

A Marine-Derived, Multi-mineral Supplement Influences Bacterial Fermentation and Short Chain Fatty Acid Profile In Vitro.

Aquamin is a calcium-rich multi-mineral supplement derived from the red marine algae, Lithothamnion species. Calcium supplementation has been shown to exert a prebiotic-like effect on the gut microbiota and has been associated with distinct changes in lactate and short chain fatty acid (SCFA) production. Irritable bowel syndrome (IBS) subtype is associated with changes in SCFA levels compared with healthy controls. Using an ex vivo simulation model, and a fecal inoculum from a patient diagnosed with IBS, we evaluated the effects of Aquamin (at 6 and 30 mg/mL) on SCFAs and lactate production, pH and gas production, and human microbiota composition. Our results demonstrate that Aquamin increased SCFA production (acetate and propionate by 8% and 24%, respectively, at 30 mg/mL dose), significantly decreased lactate production (30 mg/mL), and increased colonic fluid pH without inducing changes in colonic gas production or gastrointestinal (GI) microbiota composition. These results indicate that Aquamin may play a role in optimizing GI microbial function in an ex vivo setting.

Carotenoids: Considerations for Their Use in Functional Foods, Nutraceuticals, Nutricosmetics, Supplements, Botanicals, and Novel Foods in the Context of Sustainability, Circular Economy, and Climate Change.

Carotenoids are versatile isoprenoids that are important in food quality and health promotion. There is a need to establish recommended dietary intakes/nutritional reference values for carotenoids. Research on carotenoids in agro-food and health is being propelled by the two multidisciplinary international networks, the Ibero-American Network for the Study of Carotenoids as Functional Foods Ingredients (IBERCAROT; http://www.cyted.org) and the European Network to Advance Carotenoid Research and Applications in Agro-Food and Health (EUROCAROTEN; http://www.eurocaroten.eu). In this review, considerations for their safe and sustainable use in products mostly intended for health promotion are provided. Specifically, information about sources, intakes, and factors affecting bioavailability is summarized. Furthermore, their health-promoting actions and importance in public health in relation to the contribution of reducing the risk of diverse ailments are synthesized. Definitions and regulatory and safety information for carotenoid-containing products are provided. Lastly, recent trends in research in the context of sustainable healthy diets are summarized.

Growth inhibitory effects of casein hydrolysates on human cancer cell lines.

The aim of this study was to investigate the effects of unhydrolysed/intact casein and eight different sodium casein hydrolysates (a-h) on the viability and growth of human cancer cell lines. Both human Jurkat T cells and Caco-2 cells were incubated with increasing concentrations of the test compounds (0.5-10% v/v) for 24 h. Cell viability was assessed using the MTT, lactate dehydrogenase (LDH) release and Trypan Blue assays. Cell growth was monitored using the MTT, Trypan Blue and Bromodeoxyuridine (BrdU) proliferation assays. Casein hydrolysates b, c and f had an inhibitory effect on the viability and growth of both cell lines. The casein hydrolysates did not negatively affect the membrane integrity of both Jurkat and Caco-2 cells. In Jurkat cells hydrolysates a and h had an inhibitory effect on DNA synthesis after 24 h, while in Caco-2 cells DNA synthesis was not affected. In conclusion, we found that the different casein hydrolysates had cell-specific effects which target particular functions within the cell. Overall, casein hydrolysates had no effect on membrane integrity while they had varied effects on mitochondrial activity and DNA synthesis in the different cell lines.

Carotenoid content of commonly consumed herbs and assessment of their bioaccessibility using an in vitro digestion model.

Herbs are a rich source of bioactive phytochemicals such as carotenoids, which are known to exert various positive biological effects. However, there is very limited information in the literature regarding the content and bioavailability of carotenoids from commonly consumed herbs. Therefore, the objectives of the present study were first, to determine the carotenoid content of eight herbs namely basil (Ocimum basilicum), coriander (Coriandrum sativum), dill (Anethum graveolens), mint (Metha L.), parsley (Petroselinum crispum), rosemary (Rosmarinus officinalis), sage (Salvia officinalis), and tarragon (Artemisia dracunculus L.); and second, to assess carotenoid bioaccessibility from these herbs using a simulated human in vitro digestion model. Carotenoid bioaccessibility is defined as the amount of carotenoids transferred to micelles after digestion when compared with the original amount present in the food. The content of individual carotenoids varied significantly among the herbs tested. Carotenoid bioaccessibility varied from 0 to 42.8%. Basil and coriander, and their respective micelles, contained the highest levels of beta-carotene, beta-cryptoxanthin, and lutein + zeaxanthin. Our findings show that herbs are rich sources of carotenoids and that these foods can significantly contribute to the intake of bioaccessible carotenoids.

The role of calcium in apoptosis induced by 7beta-hydroxycholesterol and cholesterol-5beta,6beta-epoxide.

Oxysterols, such as 7beta-hydroxy-cholesterol (7beta-OH) and cholesterol-5beta,6beta-epoxide (beta-epoxide), may have a central role in promoting atherogenesis. This is thought to be predominantly due to their ability to induce apoptosis in cells of the vascular wall and in monocytes/macrophages. Although there has been extensive research regarding the mechanisms through which oxysterols induce apoptosis, much remains to be clarified. Given that experimental evidence has long associated alterations of calcium (Ca(2+)) homeostasis to apoptotic cell death, the aim of the present study was to determine the influence of intracellular Ca(2+) changes on apoptosis induced by 7beta-OH and beta-epoxide. Ca(2+) responses in differentiated U937 cells were assessed by epifluorescence video microscopy, using the ratiometric dye fura-2. Over 15-min exposure of differentiated U937 cells to 30 muM of 7beta-OH induced a slow but significant rise in fura-2 ratio. The Ca(2+) channel blocker nifedipine and the chelating agent EGTA blocked the increase in cytoplasmic Ca(2+). Moreover, dihydropyridine (DHP) binding sites identified with BODIPY-FLX-DHP were blocked following pretreatment with nifedipine, indicating that the influx of Ca(2+) occurred through L-type channels. However, following long-term incubation with 7beta-OH, elevated levels of cytoplasmic Ca(2+) were not maintained and nifedipine did not provide protection against apoptotic cell death. Our results indicate that the increase in Ca(2+) may be an initial trigger of 7beta-OH-induced apoptosis, but following chronic exposure to the oxysterol, the influence of Ca(2+) on apoptotic cell death appears to be less significant. In contrast, Ca(2+) did not appear to be involved in beta-epoxide-induced apoptosis.

Investigation of beta-carotene and lutein transport in Caco-2 cells: carotenoid-carotenoid interactions and transport inhibition by ezetimibe.

Carotenoid bioavailability is influenced by a number of factors including the presence of other carotenoids, which may enhance or inhibit the transport of one another by intestinal cells. Therefore, the objectives of the present study were: first, to determine carotenoid uptake and secretion (i. e. transport) by supplementing differentiated Caco-2 cells with increasing concentrations of either lutein or beta-carotene (0 - 1 microM); second, to assess any interactions between beta-carotene and lutein on their cellular uptake and secretion; and third, to a minor extent, to determine the effects of a carotenoid absorption inhibitor, ezetimibe, on beta-carotene and lutein transport. The carotenoid mixes were used at molar ratios of 1:1 and 4:1. At equimolar concentrations, lutein had a negative impact on beta-carotene transport and vice versa. However, these effects were not seen when the ratios were adjusted to 4:1. Following treatment with ezetimibe (25 - 100 microM) for 16 hours there was a reduction in beta-carotene transport, whereas a non-significant reduction in lutein transport was observed. In conclusion, this study confirmed that beta-carotene and lutein interact during their absorption, depending on the concentration/ratios used, and that carotenoid absorption is partially affected by ezetimibe.

Development of a dehydrated fortified food base from fermented milk and parboiled wheat, and comparison of its composition and reconstitution behavior with those of commercial dried dairy-cereal blends.

Dehydrated blends of milk and cereal are reconstituted and consumed as a nutritious soup or porridge in many regions; the composition and reconstitution behavior of the blends are likely to impact on nutritional quality and consumer acceptability of the soup/porridge. Experimental samples of dried fermented milk-bulgur wheat blend (FMBW) and commercial samples of dried dairy-cereal blends, namely kishk, tarhana, and super cereal plus corn-soy blend (SCpCSB) were compared for composition, color, water sorption, and reconstitution characteristics. FMBW blends had higher contents of protein, Ca, lactose and lactic acid, lower levels of salt (NaCl) and Fe, and a lighter, more-yellow color (higher L* and b*-color co-ordinates) than tarhana or kishk. Compared with SCpCSB, FMBW had numerically higher levels of protein, lactose, and lactic acid, lower levels of Ca, Fe, Zn, and Mg, and lower pH. Tarhana had highest mean levels of starch, and on reconstitution (133 g/kg) had highest water holding capacity, viscosity during pasting and cooling, yield stress (σ 0), consistency coefficient (K), and viscosity on shearing from 20 to 120 s-1 at 60°C. Reconstituted FMBW, kishk, and SCpCSB had similar pasting and flow behavior properties. Overall, the composition (starch, protein, Ca, Mg), pasting and flow behavior characteristics of FMBW were closer to those SCpCSB and kishk than to tarhana. The results suggest that the FMBW powder, on appropriate supplementation with Ca, Fe, Zn and Mg, could be used for the development of customized fortified blended foods for specific groups.

Characterisation of the in vitro bioactive properties of alkaline and enzyme extracted brewers' spent grain protein hydrolysates.

Brewer's spent grain (BSG) is a co-product of the brewing industry that has been shown to contain a range of bioactive peptides encrypted within its protein sequences. Two methods were evaluated herein to generate bioactive peptides; (i) an alkaline extracted BSG protein rich fraction (BSG-PI) was hydrolysed using different combinations of proteolytic enzymes and (ii) BSG was pre-treated with carbohydrases followed by direct hydrolysis using proteolytic enzymes (BSG-DH). BSG-DH with Alcalase/Flavourzyme resulted in significantly higher (p < .05) protein yield when compared to BSG-PI (63.09 ±â€¯0.27 and 58.90 ±â€¯1.45%, respectively). The antioxidant activities (ORAC, FRAP and ABTS) of the BSG-PI and -DH hydrolysates differed depending on the assay and proteolytic enzyme combination preparations used for hydrolysis. Inhibition of DPP-IV by the BSG-PI hydrolysates ranged from 87.01 ±â€¯0.15 to 89.61 ±â€¯0.12% while inhibition by the BSG-DH hydrolysates ranged from 35.71 ±â€¯0.72 to 85.06 ±â€¯0.17%. A significant reduction in the release of interleukin-6 in lipopolysaccharide-stimulated RAW 264.7 cells was observed following treatment with BSG-PI hydrolysates generated with Prolyve/Protease P (58.30 ±â€¯13.76%) and Corolase PP/Flavourzyme (48.02 ±â€¯10.82%) when compared to untreated LPS stimulated control cells (100%). BSG-DH hydrolysates were subjected to in vitro simulated gastrointestinal digestion (SGID) which resulted in a reduction in antioxidant activity, an increase in DPP-IV inhibition and no change in the immunomodulatory activity. Ultrafiltration of selected BSG-DH hydrolysates (through 30 and 10 kDa membranes) gave some permeates with enhanced bioactivities. The results demonstrate that direct enzymatic hydrolysis of BSG is a feasible approach for the generation of bioactive peptides without the prior use of an alkali protein extraction step.