Comparison of a modified spectrophotometric and the pH-stat methods for determination of the degree of hydrolysis of whey proteins hydrolysed in a tangential-flow filter membrane reactor

A procedure was developed to determine the degree of hydrolysis (DH) of whey protein hydrolysates (WPH) during hydrolysis in either 3 kDa or 10 kDa tangential-flow filter (TFF) enzymatic membrane reactors (EMR). Protease N Amano G (IUB 3.4.24.28, Bacillus subtilis) was used to hydrolyse an initial 5% (w v^?1) aqueous solution of whey protein isolate (86.98% protein) at pH 7.0 and 55 °C with continuous recirculation and simultaneous removal of hydrolysates through the TFF, in single- or two-stage operation. The DH in the permeate and the retentate were determined as the concentration of the free α-NH₂ using 2,4,6-trinitrobenzene 1-sulphonic acid (TNBS) and compared to the pH-stat method. In the new method, the DH of the permeate, the retentate and for the total EMR process could be quantified together or independently. The pH-stat method exaggerated the DH in the EMR because of the leakage of the alkali. The TNBS method was more reliable for DH estimation in the EMR.     

Kinetics of the inhibition of renin and angiotensin I-converting enzyme by flaxseed protein hydrolysate fractions

Enzymatic hydrolysates from flaxseed protein were investigated for in vitro inhibition of angiotensin I-converting enzyme (ACE) and renin activities. Pepsin, ficin, trypsin, papain, thermolysin, pancreatin and Alcalase were used to hydrolyze flaxseed proteins followed by fractionation using ultrafiltration to isolate low-molecular-weight peptides, and separation of the Alcalase hydrolysate into cationic peptide fractions. Using N-(3-[2-furyl]acryloyl)-phenylalanylglycylglycine as substrate, the protein hydrolysates showed a concentration-dependent ACE inhibition (IC₅₀, 0.0275–0.151 mg/ml) with thermolysin hydrolysate and Alcalase cationic peptide fraction I (FI) showing the most potent activity. Flaxseed peptide fractions also showed no or moderate inhibitory activities against human recombinant renin (IC₅₀, 1.22–2.81 mg/ml). Kinetics studies showed that the thermolysin hydrolysate and FI exhibited mixed-type pattern of ACE inhibition whereas cationic peptide fraction II inhibited renin in uncompetitive fashion. These results show that the protein components of flaxseed meal possess peptide amino acid sequences that can be exploited as potential food sources of anti-hypertensive agents.     

Bioactive peptides generated in an enzyme membrane reactor using Bacillus lentus alkaline peptidase

Bacillus lentus alkaline peptidase (BLAP) was used for casein (CN) hydrolysis in an enzyme membrane reactor (EMR) because it was found that BLAP was competitively inhibited by its products. The employed membranes had different molecular weight cut-offs (MWCO 1, 5 and 10 kDa). It was shown that the productivity of the EMR could be significantly improved (28 %) in comparison with batch hydrolysis under the same conditions after 20 h. All resulting EMR peptide mixtures showed a homogenous peptide pattern in HPLC–UV analysis. The obtained peptide mixtures exhibited Angiotensin-I-converting enzyme (ACE) inhibitory and antioxidative activity. The ACE inhibition of the peptide mixtures was dependent on the MWCO of the membranes. The resulting apparent IC₅₀ values were 115, 131 and 420 μg ml^?1 for the 1, 5 and 10 kDa MWCO membranes, respectively. In kinetic studies, a mixed-type inhibition was observed for the three peptide mixtures. The radical scavenging activity was determined with the ABTS assay, and IC₅₀ values between 20 and 25 μg ml^?1 were obtained for the generated peptide mixtures. In addition, the identified VYPFPGPIPN peptide exhibited ACE inhibition and antioxidant activity with IC₅₀ values of 325 and 6.2 μM, respectively. The peptide YQEPVLGPVRGPFPIIV exhibited radical scavenging activity with an IC₅₀ value of 5.2 μM.     

Preparation and identification of angiotensin I-converting enzyme inhibitory peptides from sweet potato protein by enzymatic hydrolysis under high hydrostatic pressure

Sweet potato protein hydrolysates (SPPH) with angiotensin I-converting enzyme (ACE) inhibitory activity were prepared by papain, pepsin and alcalase under high hydrostatic pressure (HHP, 100–300 MPa). HHP significantly increased degree of hydrolysis (DH), nitrogen recovery (NR) and molecular weight (MW) <3 kDa fractions contents of SPPH by all three enzymes (P < 0.05). MW < 3 kDa peptide fractions from SPPH by alcalase under 100 MPa showed the highest ACE inhibitory activity (IC₅₀ value 32.24 µg mL⁻¹), and was subjected to purification and identification by semi-preparative RP-HPLC and LC-MS/MS. Fifty-four peptides ranged from 501.28 to 1958.88 Da with 5–18 amino acids were identified and matched sporamin A and B sequences. Five identified peptides with sequences of VSAIW, AIWGA, FVIKP, VVMPSTF and FHDPMLR displayed good ACE inhibitory activity with the contribution of Val, Trp, Phe and Arg. Thus, SPPH by enzymatic hydrolysis under HHP can be potentially used in functional food.     

Angiotensin-I-converting enzyme inhibitory activity and bitterness of enzymatically-produced hydrolysates of shrimp (Pandalopsis dispar) processing byproducts investigated by Taguchi design

The importance of water-to-substrate ratio, protease type, percent enzyme and incubation time on hydrolysates produced from shrimp processing byproducts was investigated using Taguchi’s L₁₆ (4⁵) experimental design. Protease type significantly (p < 0.05) influenced soluble yield, degree of hydrolysis (DH), angiotensin-I-converting enzyme (ACE) inhibitory activity and bitterness of hydrolysates, while percent enzyme only affected the DH. Hydrolysates produced by Alcalase and Protamex possessed strong ACE inhibitory activity (IC₅₀ = 100–200 μg/ml and 70 μg/ml, respectively), accompanied by high yield, high DH and strong bitterness. Furthermore, ACE inhibition was positively correlated (r² = 0.87) with bitterness of the hydrolysates. Fractionation by size-exclusion chromatography revealed that the bitter substances, which also showed strong ACE inhibition, were <3 kDa in size and contained many hydrophobic residues, including Tyr, Phe, Leu, Ile, Val and Lys. Despite the bitterness, these hydrolysates may have potential health benefits, arising from their potent ACE inhibitory activity.     

Pilot-scale production of hydrolysates with altered bio-functionalities based on thermally-denatured whey protein isolate

Whey protein isolate (WPI) solutions (100 g L⁻¹ protein) were subjected to a heat-treatment of 80 °C for 10 min. Unheated and heat-treated WPI solutions were hydrolysed with Corolase^® PP at pilot-scale to either 5 or 10% degree of hydrolysis (DH). Hydrolysates were subsequently processed via cascade membrane fractionation using 0.14 μm, and 30, 10, 5 and 1 kDa cut-off membranes. The compositional and molecular mass distribution profiles of the substrate hydrolysates and membrane processed fractions were determined. Whole and fractionated hydrolysates were assayed for both angiotensin-I-converting enzyme (ACE) inhibitory activity and ferrous chelating capabilities. A strong positive correlation (P < 0.01) was established between the average molecular mass of the test samples and the concentration needed to chelate 50% of the iron (CC₅₀) in solution. The lowest ACE inhibition concentration (IC₅₀ = 0.23 g L⁻¹ protein) was determined for the 1 kDa permeate of the heat-treated 10% DH hydrolysate.     

Angiotensin I-converting enzyme inhibitory activity of chickpea and pea protein hydrolysates

ACE inhibitory activity was studied for different hydrolysates obtained from protein concentrates of chickpea (kabuli and desi) and yellow pea (Golden) using in vitro gastrointestinal simulation, alcalase/flavourzyme, and papain. Protein/peptide profiles studied by SDS–PAGE and SE-HPLC, showed a rich composition of the hydrolysates in small peptides having MWs under 4 kDa. Papain hydrolysed yellow pea proteins showed the highest ACE inhibitory activity. In addition, chickpea desi proteins hydrolysed by in vitro gastrointestinal simulation showed higher ACE inhibition (IC₅₀ of 140 ± 1 μg/ml) compared to its digests obtained by alcalase/flavourzyme (IC₅₀ of 228 ± 3 μg/ml) or papain (IC₅₀ of 180 ± 1 μg/ml) and to chickpea kabuli hydrolysed by gastrointestinal simulation (IC₅₀ of 229 ± 1 μg/ml). The results demonstrate that enzymatic hydrolysates of chickpea and pea proteins contain bioactive ACE inhibitory peptides; furthermore, the type of enzyme used for hydrolysis affects the ACE inhibitory activity.     

Evaluation of angiotensin I-converting enzyme (ACE) inhibitory activities of smooth hound (Mustelus mustelus) muscle protein hydrolysates generated by gastrointestinal proteases: identification of the most potent active peptide

In this study, smooth hound protein hydrolysates (SHPHs), obtained by treatment with various gastrointestinal proteases, were analyzed for their angiotensin I-converting enzyme (ACE) inhibitory activities. Protein hydrolysates were obtained by treatment with crude alkaline enzyme extract, low molecular weight (LMW) alkaline protease, trypsin-like protease and pepsin from Mustelus mustelus, and bovine trypsin. All hydrolysates exhibited inhibitory activity toward ACE. Hydrolysate generated with alkaline protease extract displayed the highest ACE inhibitory activity, and the higher inhibition activity (82.6% at 2 mg/mL) was obtained with a hydrolysis degree of 18.8%. This hydrolysate was then fractionated by size exclusion chromatography on a Sephadex G-25 into five major fractions (P₁–P₅). ACE inhibitory activities of all fractions were assayed, and P₃ was found to display a high ACE inhibitory activity (62.24% at 1 mg/mL). P₃ was then fractionated by reversed-phase high-performance liquid chromatography (RP-HPLC) and ten fractions of ACE inhibitors were found (F₁–F₁₀). Sub-fraction F₃ showed the strongest ACE inhibitory activity, being able to suppress more than 60% of initial enzyme activity at a concentration of 100 μg/mL. The amino acid sequence of peptide F₃ was determined by ESI/MS and ESI–MS/MS as Ala-Gly-Ser, and the IC₅₀ value for ACE inhibitory activity was 0.13 ± 0.03 mg/mL. Further, purified peptide F₃ maintained inhibitory activity even after in vitro digestion with gastrointestinal proteases in order to demonstrate gastrointestinal stability digestion to enable oral application. These results indicate that smooth hound protein hydrolysate possesses potent antihypertensive activity.     

Chemical and functional characterization of Gum karaya (Sterculia urens L.) seed meal

Chemical and functional characterization of the meal of Gum karaya (Sterculia urens) whole and dehulled–defatted seed were investigated. Dehulled–defatted seed meal (DDSM) contained 40.7% of protein as compared to 20.4% in the whole seed meal (WSM). Essential minerals of Ca 39.5, Fe 4.4, K 9.6 and P 995 mg/100 g were found in DDSM. A solid to solvent ratio 1:40 and extraction time of 60 min was optimum. The highest protein solubility (93.97%) was observed at pH 12 and the lowest (26.41%) protein solubility at pH 6. The solubility of protein was enhanced in the presence of 0.1 M and 0.5 M NaCl between pH rang of 2–12. The protein precipitation from an alkaline extract of pH 10 was maximum (81%) at pH 6. Polyacrylamide gel electrophoresis shows that 14 bands of SDS soluble protein subunits were observed in WSM and DDSM. A higher buffer capacity of the meal was observed in the acid medium than in alkaline medium. The DDSM had an initial moisture content (IMC) of 5.16%, which equilibrated at 32% RH and moisture content increased sharply at higher humidity (70%) which, indicated that the non-hygroscopic in nature. Water holding capacity and oil holding capacity were 111 and 81, 67 and 114 g/100 g of WSM and DDSM respectively. DDSM showed very good foam capacity (32%) and stability (75%) even after 90 min at room temperature. The emulsification capacity of DDSM was found to be 20 mL/g sample.     

Identification and quantification of phenolic compounds from sunflower (Helianthus annuus L.) kernels and shells by HPLC-DAD/ESI-MS

Phenolic compounds were extracted from defatted sunflower (Helianthus annuus L.) kernels and shells and characterised by HPLC with diode array and electrospray ionisation (ESI) mass spectrometric detection in the negative mode. Quantification of individual compounds was carried out by external calibration. Among the eleven compounds analysed 5-O-caffeoylquinic acid was predominant amounting up to 59.1 mg/100 g in the shells and 3050.5 mg/100 g in the kernels. The specific fragmentation patterns of mono- and dihydroxycinnamoylquinic acids allowed the unambiguous distinction of several stereoisomers which have not been described for sunflower seeds and seed shells so far. The total phenolic content of about 4200 mg/100 g on a dry matter basis revealed defatted sunflower meal to be a promising source of phenolic compounds that might be recovered and used as natural antioxidants. Furthermore, the press residues originating from sunflower oil extraction were shown to be still rich in phenolic antioxidants, thus, providing the opportunity to valorize these by-products in terms of sustainable agricultural production.     

Improvement of emulsifying properties of Maillard reaction products from β-conglycinin and dextran using controlled enzymatic hydrolysis

A novel emulsifier was prepared by conjugating soy β-conglycinin and dextran (MW 67 kDa) under dry-heated Maillard reaction followed by trypsin hydrolysis with the degree of hydrolysis (DH) at 2.2% and 6.5%. The emulsifying properties of β-conglycinin, β-conglycinin–dextran conjugates and hydrolysates of β-conglycinin–dextran conjugates (DH 2.2% and DH 6.5%) were investigated using zeta-potential, droplet size and creaming index of the emulsions. The results showed that hydrolysates of β-conglycinin–dextran conjugates (DH 2.2%) were capable of forming a fine emulsion (d₄₃ = 0.62 ± 0.04 μm, pH 7.0) which remained stable during 4 weeks of storage. A variety of physicochemical and interfacial properties of β-conglycinin, β-conglycinin–dextran conjugates and hydrolysates of β-conglycinin–dextran conjugates were investigated. Hydrolysates of β-conglycinin–dextran conjugates (DH 2.2%) had a much higher fraction of protein adsorption (F_ads) and a significantly lower saturation surface load (Γ_sat) compared with β-conglycinin, β-conglycinin–dextran conjugates and hydrolysates of β-conglycinin–dextran conjugates (DH 6.5%). This might be due to its higher molecular flexibility, which benefited the adsorption and unfolding of peptide molecules at the droplet interface. These might explain its markedly improved emulsifying capability. The conjugation of β-conglycinin and dextran effectively enhanced the hydrophilicity of the oil droplets surfaces and improved the steric repulsion between the oil droplets. Therefore the emulsions were still stable after 4 weeks of storage against pH, ionic strength and thermal treatment. This study demonstrated that controlled enzymatic hydrolysis of protein–polysaccharide conjugates could be an effective method for preparing favourable emulsifiers.     

Continuous production of tempe-based bioactive peptides using an automated enzymatic membrane reactor

The utilization of bioactive peptides from food-grade raw materials has received increasing interest, especially for the production of functional foods. Within this study, a combination of fermentation and in vitro papain hydrolysis was devised to produce bioactive peptides exhibiting antioxidant property and angiotensin I-converting enzyme (AICE) inhibitory activity. The utilization of an automated membrane reactor system developed in this study could facilitate the continuous hydrolysis of tempe flour-rich in protein under constant flux, thus constant residence time operation. The optimum operating conditions were: [E]/[S] = 10% (w/v) and τ = 9 h. With a 10-kDa membrane filtration, the resulted antioxidant capacity and AICE inhibition were 0.033 mg AEAC/mL and 50.9%, respectively. Further separation of permeate with a 2-kDa membrane cut-off leveled off the antioxidant capacity but increased the AICE inhibitory activity (i.e., Reduction of IC₅₀ of ~40%). By this, the obtained IC₅₀ for AICE was 0.08 mg protein/mL, and this value was comparable with the IC₅₀ values reported in the literature.Industrial relevanceThe production of bioactive peptides from soybean through a combination of fermentation and in vitro enzymatic hydrolysis is limitedly reported. In this work, fermented soybean (tempe) was further hydrolyzed continuously using an automated membrane reactor. The hydrolysis under optimum operating conditions could increase the antioxidant activity and AICE inhibition of permeate. This work is expected to increase the commercialization of tempe-based food products by using safer technological approaches (i.e., fermentation and membrane technology).     

Inhibition of the angiotensin-converting enzyme by grape seed and skin proanthocyanidins extracted from Vitis vinífera L. cv. País

The influence of two extracts of grape skin and seeds from Vitis vinífera L. cv. País (Chilean black grapes), rich in proanthocyanidins (PAs), was evaluated on the inhibition of the angiotensin I-converting enzyme (ACE), and the inhibition was related to the type and number of subunits of the polymeric PAs chain. Size exclusion chromatography was used to purify the extract and its characterization was made by acid catalysis depolymerization followed by HPLC. ACE activity was measured by quantitative HPLC, measuring the hyppuric acid (HA) produced from the hydrolysis of hippuryl-l-histidyl-l-leucine (HHL) by ACE. Structural compositions differed significantly between both extracts: the skin extracts do not exhibit epicatechin (EC) and epicatechin gallate (ECG), and the seed extracts did not present epigallocatechin (EGC). Skin extracts have a higher mean degree of polymerization (mDP) than seed extracts and a higher inhibition power (IC₅₀ = 0.14 ± 0.03 μM and IC₅₀ = 0.480 ± 0.03 μmol/L), respectively. The catechin (IC₅₀ = 1495 ± 90 μmol/L) and epicatechin (IC₅₀ = 1772 ± 121 μmol/L) monomers exerted lower inhibition than the either grape extract. The structural differences between grape skin and seed PAs could influence the ACE inhibition capacity. The larger inhibitory power of skin extract was associated to greater OH availability, higher mDP and the presence of EGC.     

Angiotensin I-converting enzyme (ACE) inhibitory peptide derived from Tenebrio molitor (L.) larva protein hydrolysate

Short peptides from different sources have been proven to be very efficient inhibitors of angiotensin I-converting enzyme (ACE), an enzyme with a major role in the regulation of blood pressure. In order to obtain ACE-inhibitory peptides from Tenebrio molitor (L.) larva, enzymatic hydrolysis was performed using Alcalase. The hydrolysate (DH 20 %) with the highest ACE inhibition was fractionated into four peaks according to their relative molecular weight by gel filtration on Sephadex G-15. The IC₅₀ value of peak 2 (with the relative molecular weight 180–500 Da) was 0.23 mg/mL, while it was 0.39 mg/mL of hydrolysate before fractionation determined by reversed-phase high performance liquid chromatography (RP-HPLC). Antihypertensive activity of peak 2 was proven by single oral administration to spontaneously hypertensive rats (SHR). Multiple dose oral administration (100, 200, and 400 mg/kg body weight) to SHR led to a significant decrease in blood pressure for peak 2. The reduction of systolic blood pressure even reached 27 mm Hg at 4 h post-administration at a dose of 400 mg/kg body weight. Additionally, further separation and purification of the ACE-inhibitory peptides from peak 2 were performed using RP-HPLC on C₁₈ column. Then a novel peptide Tyr–Ala–Asn was identified by tandem mass spectrometry with an IC₅₀ value of 0.017 mg/mL. The result of research indicates that Tenebrio molitor (L.) larva protein may be a suitable candidate to prepare ACE-inhibitory peptides which could be used for food industry and nutraceuticals.     

Oil content and fatty acid composition of some underutilized legumes from Nigeria

Three underutilized legumes from Nigeria, Brachystegia eurycoma, Tamarindus indica and Mucuna flagellipes, have been subjected to standard analytical techniques in order to evaluate proximate composition, physicochemical properties and contents of nutritional valuable elements and fatty acids of the seeds and oils. The proximate analysis indicated that the oil content was 5.87 ± 0.30, 7.20 ± 0.45 and 3.77 ± 0.21 g/100 g for B. eurycoma, T. indica and M. flagellipes, respectively. The seeds are rich in protein and carbohydrate, the protein content ranging from 11.82 ± 0.25 g/100 g–24.94 ± 0.18 g/100 g dry matter. These compare favourably with high protein animal sources like oyster, beef, pork and marine fishes. The iodine value of two of the oils place them in the non-drying group of oils, while the composition of all the oils compare well with those of rape seed, sesame, sunflower and groundnut seed oils. This suggests their use as edible oils.Analyses of the oils for fatty acids indicate that the oils contain linoleic acid which is one of the three essential fatty acids. The dominant fatty acids however are linoleic, palmitic, oleic and stearic acids with oleic acid having the highest percentage 24.13–31.50%. Eight nutritional valuable minerals were determined in the seed flours. The seeds are rich in potassium 52.1 mg/100 g–131 mg/100 g. They also contain significant concentration of iron 4.55 mg/100 g–8.20 mg/100 g.     

Inhibition of angiotensin I-converting enzyme by wheat gliadin hydrolysates

A tryptic gliadin hydrolysate was fractionated into peptide fractions, which were assigned to either the central domain (CD) or terminal domains (TD) of gliadins. The domains were expected to contain amino acid (AA) sequences which, when released from the parent protein, inhibit the angiotensin I-converting enzyme (ACE), which plays a key role in regulating blood pressure. A proline (Pro) poor TD related fraction, containing the smallest peptides, showed the highest ACE inhibitory activity (IC₅₀ = 0.33 mg/ml). Additional peptidases were selected based on their in silico predicted ability to release ACE inhibitory peptides. Further hydrolysis of the tryptic hydrolysate fractions with thermolysin, Clarex, Alcalase and Esperase increased ACE inhibitory activities. Immobilised Ni²⁺-ion affinity chromatography (IMAC) purification of a TD related peptide fraction obtained by sequential hydrolysis with trypsin and thermolysin yielded a fraction with an IC₅₀ value of 0.02 mg/ml. This IMAC fraction was enriched in histidine and hydrophobic AA (Pro, Val, Ile, Leu and Phe).     

Preparation and antihypertensive activity of peptides from Porphyra yezoensis

This research was to develop a antihypertensive peptide, an efficient angiotensin converting enzyme (ACE) inhibitor (ACEI), from Porphyra yezoensis. Seven commercial enzymes were screened and then enzymatic hydrolysis conditions were optimised. The results showed that alcalase was the most effective for hydrolysis and its optimum conditions for achieving the highest antihypertensive activity of peptide were 1.5% substrate, 5% alcalase, pH 9.0, and temperature of 50 °C at a hydrolysis time of 60 min. The antihypertensive peptide produced under the optimum conditions had a high ACE inhibition rate of 55.0% and a low IC₅₀ value of 1.6 g/l and remained at high stability at temperatures of 4, 25, and 37 °C, pH values of 2.0 and 8.0, and after pepsin and trypsin treatments. Major proteins from P. yezoensis were glutelin, albumin, and gliadin. The albumin and glutelin had higher hydrolysis rates than the gliadin, but the IC₅₀ value of glutelin was the lowest, which indicated that the antihypertensive peptide from glutelin was more functional.     

Pilot-scale production of low molecular weight peptides from corn wet milling byproducts and the antihypertensive effects in vivo and in vitro

A pilot-scale production was developed to enhance the value of proteins from corn gluten meal (CGM). Corn protein isolate (CPI) with high protein content (90.68%) was obtained through heat treatment of CGM (150 kg) in aqueous alkaline solution. Two-step enzymatic hydrolysis and multistage separation were applied to enrich corn oligopeptides (COP) with low molecular weights, 96.77% of which were less than 1000 Da. The greatest antihypertensive effect of COP treatment in spontaneously hypertensive rats (SHR) was observed at a dose of 0.45 g/kg. One major ACE-inhibitory peptide, Ala-Tyr, was identified and quantified (9.16 ± 0.08 mg/g) from COP. The ACE inhibitory activity of Ala-Tyr (IC₅₀ = 0.037 mg/ml) was over 27 times higher than that of COP (IC₅₀ = 1.020 mg/ml). The results indicate that COP may be a source of natural antihypertensive compounds that could be used for drugs or functional food ingredients.     

Separation of angiotensin I‐converting enzyme inhibitory peptides from bovine connective tissue and their stability towards temperature,pH and digestive enzymes

Bovine collagen was isolated from connective tissue, a by‐product in the meat processing industry and characterised by SDS‐PAGE. Alcalase and papain were employed to generate collagen hydrolysates with different degree of hydrolysis (DH). In vitro angiotensin I‐converting enzyme (ACE) inhibitory activities were evaluated and the two most potent hydrolysates from each enzyme were separated by two‐step purification. Both alcalase‐catalysed and papain‐catalysed hydrolysates exhibited strong ACE inhibitory capacities with IC₅₀ values of 0.17 and 0.35 mg mL^?1, respectively. Purification by ion‐exchange chromatography and gel filtration chromatography revealed higher ACE inhibitory activities in one fraction from each enzyme with IC₅₀ values of 3.95 and 7.29 μg mL^?1. These peptide fractions were characterised as 6‐12 amino acid residues by MALDI‐TOF/MS. The peptides retained their activity (>90%) after exposure to processing temperature and pH and in vitro simulated gastrointestinal digestion. The present results demonstrated that collagen peptides can be utilised for developing high value‐added ingredients, for example ACE inhibitory peptides.     

Composition and antioxidative activities of supercritical CO2-extracted oils from seeds and soft parts of northern berries

The present study investigated the composition and the antioxidative activities of oils from the seeds and the soft parts of a range of northern berries extracted by supercritical CO₂. The seed oils of the species of Rubus, Vaccinium, Empetrum, Fragaria and Hippophaë were rich in linoleic (18:2n-6, 34-55% of total fatty acids) and ??-linolenic (18:3n-3, 29-45% of total) acids with n-6:n-3 ratios of 1:1-1:2. The seed oils of the species Ribes contained, in addition to linoleic and ??-linolenic acids, ??-linolenic (18:3n-6) and stearidonic (18:3n-4) acids. In seed oils from European rowanberry (Sorbus aucuparia L.) and snowball berry (Viburnum opulus L.), linoleic and oleic (18:1n-9) acids together exceeded 90% of the total fatty acids. The sea buckthorn (SB) pulp oil had palmitoleic (16:1n-7), palmitic (16:0) and oleic acids as the major fatty acids. The SB pulp oil and snowball berry seed oil were rich in ??-tocopherol (120 and 110 mg/100 g oil, respectively), whereas raspberry seed oil contained a high level of ??-tocopherol (320 mg/100 g oil). Seed oils of cranberry (180 mg/100 g oil), Arctic cranberry (190 mg/100 g oil) and lingonberry (120 mg/100 g oil) are rich sources of ??-tocotrienol. The berry seed oils and the SB pulp oil showed varying peroxyl radical scavenging efficacies (300-2300 ??mol ??-tocopherol equivalent per 100 g oil) and inhibitory effects on perioxidation of microsomal lipids (250-1200 ??mol trolox equivalent per 100 g oil) in vitro. The peroxyl radical scavenging activity positively correlated with the total content of tocopherols and tocotrienols of the oils (r = 0.875, P = 0.001). The SB seed oil and pulp oil were active in scavenging superoxide anions produced by xanthine-xanthine oxidase system and inhibited Cu²⁺-induced LDL oxidation in vitro. The SB oils also protected purified DNA and rat liver homogenate from UV-induced DNA oxidation in vitro. The current research suggests potential of supercritical CO₂-extracted oils from northern berries as nutraceuticals and ingredients of functional foods.