Antioxidative and functional properties of protein hydrolysate from defatted skipjack (Katsuwonous pelamis) roe

Antioxidative and functional properties of protein hydrolysate from defatted skipjack (Katsuwonous pelamis) roe, hydrolysed by Alcalase 2.4 L (RPH) with different degrees of hydrolysis (DH) at various concentrations were examined. As DH increased, the reduction of DPPH, ABTS radicals scavenging activities and reducing power were noticeable (p < 0.05). The increases in metal chelating activity and superoxide scavenging activity were attained with increasing DH (p < 0.05). However, chelating activity gradually decreased at DH above 30%. All activities except superoxide anion radical scavenging activity increased as the concentration of hydrolysate increased (p < 0.05). Hydrolysis using Alcalase could increase protein solubility to above 80% over a wide pH range (2–10). The highest emulsion ability index (EAI) and foam stability (FS) of hydrolysates were observed at low DH (5%) (p < 0.05). Concentrations of hydrolysates determined interfacial properties differently, depending on DH. The molecular weight distribution of RPH with 5%DH (RPH5) was determined using Sephadex G-75 column. Two major peaks with the molecular weight of 57.8 and 5.5 kDa were obtained. Fraction with MW of 5.5 had the strongest metal chelating activity and ABTS radical scavenging activity. The results reveal that protein hydrolysates from defatted skipjack roe could be used as food additives possessing both antioxidant activity and functional properties.     

Comparison of in vitro digestion characteristics and antioxidant activity of hot- and cold-pressed peanut meals

Due to the poor protein solubility, hot-pressed peanut meal (HPM) has less value than cold-pressed peanut meal (CPM) in the food industry. The objective of this study was to determine whether the denatured proteins in HPM were suitable for hydrolysis by digestive enzymes. The hydrolysis characteristics and antioxidant activity of HPM and CPM during in vitro digestion were compared. The results showed that HPM was hydrolysed more extensively than CPM. There were more free amino acids and small peptides with MW < 5 kDa in HPM hydrolysates. In addition, HPM hydrolysates displayed stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,20-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging activities, ORAC value and reducing power than CPM hydrolysates, which could be due to the higher content of small peptides, antioxidative amino acids and melanoidins in HPM hydrolysates. The above results indicated that HPM was a potential source of protein supplement for human consumption.     

Functionalities and antioxidant properties of protein hydrolysates from the muscle of ornate threadfin bream treated with pepsin from skipjack tuna

Functional properties and antioxidant activities of protein hydrolysates prepared from ornate threadfin bream (Nemipterus hexodon) muscle, using skipjack tuna pepsin, with different degrees of hydrolysis (DH: 10%, 20% and 30%), were evaluated. Emulsifying and foaming properties of hydrolysates were governed by their DH and concentrations used. Hydrolysates with 20% DH had the highest scavenging activities for ABTS and DPPH radicals. However, chelating activity of hydrolysates for ferrous ion increased as DH increased. Size exclusion chromatography of the hydrolysate with 20% DH using Sephadex G-25 revealed that antioxidative peptides with molecular weight of approximately 1.3 kDa exhibited the highest ABTS radical-scavenging activity. In vitro simulated gastrointestinal digestion indicated that ABTS radical-scavenging activity of the antioxidative peptides was not affected by pepsin hydrolysis, whilst further digestion by pancreatin enhanced the activity. Therefore, protein hydrolysate from the muscle of ornate threadfin bream produced by skipjack tuna pepsin can be used as a promising source of functional peptides with antioxidant properties.     

Oxidative stability of mahi mahi red muscle dipped in tilapia protein hydrolysates

The oxidative stability of mahi mahi red muscle dipped in tilapia protein hydrolysates was evaluated. Alkali solubilised tilapia protein isolate was hydrolysed using Flavourzyme to 13% degree of hydrolysis. Whole tilapia protein hydrolysate and ultrafiltered fraction <10 kDa were used as dip solutions. Mahi mahi red muscle was dip treated either for 2 or 4 min and stored at 4 °C. Lipid hydroperoxides (PV), thiobarbituric acid reactive substances (TBARS) and a∗ value were measured at regular intervals. Results showed that dip treatments significantly decreased (p < 0.05) the formation of PV and TBARS over 90 h storage time. There was no significant (p > 0.05) difference between WH and <10 kDa fractions, and between 2 and 4 min treatments. Red colour of treated samples measured as a∗ value decreased with storage time, but was not significantly different from the control. It could be concluded that dip treatment for 2 min in whole tilapia protein hydrolysate may be used as a potential antioxidant treatment for improving oxidative stability of fish fillets.     

Response surface optimisation for the extraction of phenolic compounds and antioxidant capacities of underutilised Mangifera pajang Kosterm. peels

The optimum extraction conditions for highest recovery of total phenolics content (TPC) and antioxidant capacities (AC) were analysed for Mangifera pajang peels (MPP), using response surface methodology. The effects of ethanol concentration (X₁: 20–80%), extraction temperature (X₂: 30–65 °C) and liquid-to-solid ratio (X₃: 20–50 mL/g) on the recovery of total phenolics (Y₁) and antioxidant capacity (Y₂) were investigated. A second order polynomial model produced a satisfactory fitting of the experimental data with regard to total phenolic content (R² = 0.9966, p < 0.0001) and antioxidant capacity (R² = 0.9953, p < 0.0001). The optimum extraction conditions for TPC were 68%, 55 °C and 32.7 mL/g, and for AC were 68%, 56 °C and 31.8 mL/g, respectively. Predicted values for extraction of TPC and AC agreed well with the experimental values. Liquid chromatography–mass spectrometry of the optimally obtained extracts from MPP revealed the major phytochemicals as mangiferin, gallic acid, catechin and epicatechin.     

Evaluation of antioxidant activities and total phenolic contents of typical malting barley varieties

Fourteen typical malting barley varieties from China were evaluated for their DPPH radical, ABTS radical cation and superoxide anion radical scavenging activities, reducing power, metal chelating activities, and total phenolic contents (TPC). All barley samples exhibited significant antioxidant activities determined by different assays, and contained significant levels of phenolic compounds. Gan4 and Wupi1 barley exhibited the highest DPPH radical scavenging activity, ABTS radical cation scavenging activity and reducing power. Gan4 and Humai16 barley showed the highest TPC, whereas the highest superoxide anion radical scavenging activity and metal chelating activity were found in Huaimai19 and Ken3 barley, respectively. The Pearson correlation analysis revealed that the TPC showed strong correlations with DPPH radical scavenging activity, ABTS radical cation scavenging activity, and reducing power (P < 0.01), whereas its correlations with superoxide anion radical scavenging activity and metal chelating activity were poor (P > 0.05). Moreover, DPPH radical scavenging activity, ABTS radical cation scavenging activity and reducing power were well positively correlated with each other (P < 0.01). Principal component analysis (PCA) was applied to understand the interrelationships among the measured antioxidant activity evaluation indices, and to gain an overview of the similarities and differences among the 14 barley varieties.     

Antioxidant and functional properties of gelatin hydrolysates obtained from skin of sole and squid

Antioxidant and functional properties were evaluated for gelatin hydrolysates obtained from sole and squid skin gelatin by Alcalase, with a degree of hydrolysis of ∼35% and ∼50%, respectively. Both hydrolysates mainly consisted of peptides below 6.5 kDa, together with peptidic material from around 16 to 6.5 kDa. Moreover, the squid hydrolysate showed a peptide band of around 26 kDa. Antioxidant properties of both gelatins were highly increased by hydrolysis, especially ABTS and metal chelating abilities. The squid hydrolysate showed the highest antioxidant capacity by FRAP, ABTS and metal chelating assays in spite of the lower content in hydrophobic amino acids. Both gelatin hydrolysates had a good solubility (over 95%). The emulsifying activity index (EAI) decreased with increasing concentration. Conversely, the foam expansion increased with increasing concentration. However, both foam and emulsion stabilities were not apparently affected by the concentration of hydrolysate. In the case of the sole hydrolysate, which showed a lower degree Pro and Lys hydroxylation, foam stability was very poor, and 50% of foam expansion was lost after 5 min at all concentrations.     

Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

Characterization of the pectic polysaccharides from pumpkin peel

The alcohol insoluble polysaccharide (AIP) was extracted from pumpkin (Cucurbita moschata Duch) peels, and they were fractionated subsequently into water soluble pectic substance (WSP), EDTA soluble pectic substance (ESP) and alkali soluble pectic substance (ASP) fractions. ASP fraction (24.78 and 46.07 g/100 g) contained the highest total sugar and uronic acid contents compared with WSP (17.78 and 4.89 g/100 g) and ESP (18.43 and 23.99 g/100 g) fractions. WSP fraction had 2 peaks with a small one close to 205 kDa and a major one close to 13 kDa, while ESP and ASP fractions had 3 peaks with higher molecular weight distributions than WSP. The sugar peaks in the ion exchange chromatography were detected in a low concentration of ammonium acetate buffer, but uronic acid peaks were detected in a high concentration of ammonium acetate buffer and 0.2 mol/l NaOH. ESP had higher glucose retardation effect than that of WSP and ASP, while WSP and ASP had higher bile acid retardation effects than that of ESP. AIP fractions activated growth in Lactobacillus brevis, Bifidobacterium bifidum and Bifidobacterium longum, whereas they inhibited growth in Escherichia coli and Clostridium perfringens in RCM medium.     

Preparation and antioxidant activity of enzymatic hydrolysates from purple sea urchin (Strongylocentrotus nudus) gonad

Purple sea urchin (Strongylocentrotus nudus) gonad was treated separately with neutral protease, papain, pepsin and trypsin. The resultant hydrolysates were fractionated using a series of ultrafiltration membranes (molecular weight cut-offs of 10, 5, 3 and 1 kDa). Five fractions were prepared from each hydrolysate and the corresponding molecular weight ranges were below 10 kDa, 5-10 kDa, 3-5 kDa, 1-3 kDa and below 1 kDa. The peptide fractions were evaluated for antioxidant activity by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reducing power assay. Results indicated that all peptide fractions possessed DPPH radical scavenging capacity and reducing power in a dose-dependent manner. For all four hydrolysates, the below 1 kDa fractions exhibited the highest DPPH radical scavenging capacity. The below 1 kDa fractions prepared with neutral protease, papain and pepsin, and the 1-3 kDa fraction prepared with trypsin showed the highest reducing capacity among corresponding hydrolysates.     

Effects of washing and membrane removal pretreatments on the antioxidant properties of grass carp (Ctenopharyngodon idella) protein hydrolysates produced by in vitro digestion

The effects of washing and membrane removal pretreatments on the antioxidant properties of grass carp protein hydrolysates prepared through in vitro digestion were investigated. Furthermore, antioxidant hydrolysate was fractionated using ultrafiltration membranes (10, 5, 3 and 1 kDa). Oxygen radical antioxidant capacity (ORAC), DPPH and ABTS‐scavenging activity in a gastrointestinal digest produced from pretreated minced grass carp was increased 1.74‐fold, 1.08‐fold and 1.72‐fold, respectively, compared to untreated minced carp. Compared to the alkaline protease hydrolysate, ORAC, ferric reducing antioxidant power, ABTS‐ and DPPH‐scavenging activity in a gastrointestinal digest prepared from pretreated minced carp were reduced by 11.5%, 60.9%, 16.3% and 78.4%, respectively. The ultrafiltration fraction (<1 kDa) displayed the highest antioxidant activity. The size of molecular weight and the amount of hydrophobic and aromatic residues in hydrolysates played an important role in antioxidant activity. Low‐molecular‐weight fish hydrolysates could serve as a potential source of functional ingredients for promoting health.     

Antioxidant activity and phenolic content of 16 raisin grape (Vitis vinifera L.) cultivars and selections

Six raisin grape cultivars and 10 new raisin grape selections were analyzed for antioxidant activity (ABTS assay) and for total and individual phenolic compounds. Samples were freeze–dried and values are reported on a dry weight basis. Antioxidant activity across the 16 samples ranged from 7.7 to 60.9 μmol Trolox/g DW, with A95-27 exhibiting the greatest activity. Total phenolic content, determined in gallic acid equivalents using the Folin–Ciocalteau assay, ranged from 316.3 to 1141.3 mg gallic acid/100 g DW and was strongly correlated (r = 0.990) with antioxidant results. Concentrations of individual phenolics were determined by HPLC. trans-Caftaric acid was the predominant compound in all samples. A95-15 contained the lowest concentration (153.5 μg/g DW) of caftaric acid, while Fiesta contained the highest concentration (598.7 μg/g DW). Selections A56-66, A95-15, and A95-27 had much higher levels of catechin (86.5–209.1 μg/g DW) and epicatechin (126.5–365.7 μg/g DW) than the other samples.     

Use of pyloric caeca extract from bigeye snapper (Priacanthus macracanthus) for the production of gelatin hydrolysate with antioxidative activity

Proteolytic activity of pyloric caeca extract (PCE) from bigeye snapper (Priacanthus macracanthus) was studied. The highest activity was observed at 55 °C and pH 8.0 when casein, N^α-Benzoyl-dl-arginine-p-nitroanilide (BAPNA) and N^α-p-Tosyl-l-arginine methyl ester hydrochloride (TAME) were used as the substrates. The activity was inhibited markedly by 1 mg/ml soybean trypsin inhibitor, whereas E-64, pepstatin A and EDTA exhibited a negligible effect on activity. The results suggested that a trypsin-like enzyme was most likely the major proteinase in PCE. As determined by activity staining, two proteolytic activity bands with apparent molecular weights of 55 and 24 kDa were found. Gelatin hydrolysate from bigeye snapper skin prepared using PCE exhibited the increases in 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities and ferric reducing antioxidative power (FRAP) as degrees of hydrolysis (DHs) increased (P < 0.05). Hydrolysates derived from gelatin using Alcalase combined with PCE showed the highest ABTS radical scavenging activity (P < 0.05). Gelatin hydrolysate prepared using Alcalase in combination with Neutrase or PCE at 500 and 1000 ppm, respectively, retarded the oxidation in both linoleic acid oxidation and lecithin liposome oxidation systems. The antioxidative peptide of gelatin hydrolysate had a molecular weight of 1.7 kDa.     

Antihypertensive effect of a bovine lactoferrin pepsin hydrolysate: Identification of novel active peptides

The potential of bovine lactoferrin (LF) as a source of antihypertensive peptides has been examined. For this purpose, LF pepsin hydrolysate with molecular mass lower than 3 kDa (LFH < 3 kDa) was prepared and orally administered to spontaneously hypertensive rats (SHR), resulting in reduced systolic blood pressure in a significant and maintained manner up to 24 h after administration. LFH < 3 kDa was further fractionated by semi-preparative high performance liquid chromatography (HPLC) and 38 peptides, contained in the active fractions, were identified by using an ion trap mass spectrometer. Based on the peptide abundance, a total of 11 peptides were chemically synthesized and their ACE inhibitory activity tested. Only three of them, corresponding to peptides of sequences LIWKL, RPYL and LNNSRAP exerted in vitro inhibitory effects on angiotensin I converting enzyme (ACE) activity and had a 50% inhibitory concentration (IC₅₀) of 0.47, 56.5 and 105.3 μM, respectively. The three peptides also showed antihypertensive effects in SHR and remarkably the effect of LIWKL remained significant for up to 24 h post-administration, similarly LFH < 3 kDa and the captopril control. The two most potent in vitro inhibitory peptides showed ex vivo inhibitory effect on ACE-dependent vasoconstriction as well. In conclusion, three novel LF-derived peptides and a pepsin LFH < 3 kDa lowered blood pressure and exhibit potential as orally effective antihypertensive compounds.     

Antioxidant properties of dried product of ‘haba-nori’, an edible brown alga, Petalonia binghamiae (J. Agaradh) Vinogradova

Dried ‘haba-nori’ Petalonia binghamiae, a brown alga, is a traditional food in the fisheries towns in Japan. To determine the antioxidant properties of the dried P. binghamiae, assays for antioxidant activities, including ferrous-reducing power, ferrous ion chelating, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and scavenging of a superoxide anion radical-generated by non-enzymatic system were tested in this study. A water extract solution contained total phenols at about 75 μmol phloroglucinol equivalents/g dry sample and showed strong antioxidant activities in the reducing power, DPPH radical and superoxide anion radical scavenging assays. The antioxidant activities were detected in high-molecular (>100 kDa), 10–30 kDa, and low-molecular (<5 kDa) fractions and were correlated with, not only phenolic compounds, but also brown compounds. The radical- scavenging activities were increased by heat treatment at 121 °C for 1 h. These results suggest that P. binghamiae is both a useful seafood and a healthy food with antioxidant activity.     

Antioxidant activity of three edible seaweeds from two areas in South East Asia

Total phenolic content (TPC) and antioxidant activity (AOA) of 50% aqueous methanol extracts of the marine algae, Padina antillarum, Caulerpa racemosa and Kappaphycus alvarezzi were studied. TPC was measured using Folin-Ciocalteu method while 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP), ferrous ion chelating (FIC) assay and beta carotene bleaching (BCB) assay were used to study their AOA. P. antillarum was found to have the highest TPC, 2430±208 mg gallic acid equivalents (GAE) per 100 g dried sample and ascorbic acid equivalent antioxidant capacity (AEAC), 1140±85 mg AA/100 g. C. racemosa and K. alvarezzi displayed lower TPC and AEAC. C. racemosa had 144±22 mg GAE/100 g dried sample of TPC and 14.3±2.0 mg AA/100 g of AEAC, while K. alvarezzi had 115±35 mg/100 g dried sample of TPC and 37.8±16.8 mg AA/100 g of AEAC. In addition, P. antillarum displayed the highest reducing power, 15.7±2.6 mg GAE/g and highest chelating ability. C. racemosa and K. alvarezzi exhibited lower reducing power, 0.737±0.423 mg GAE/g and 0.561±0.269 mg GAE/g, and lower chelating ability. However, the AOA of these three seaweeds as assessed by BCB assay were equally high.     

Purification and identification of novel antioxidant peptides from enzymatic hydrolysates of sardinelle (Sardinellaaurita) by-products proteins

In order to utilise sardinelle (Sardinellaaurita) protein by-products, which is normally discarded as industrial waste in the process of fish manufacturing, heads and viscera proteins were hydrolysed by different proteases to obtain antioxidative peptides. All hydrolysates showed different degrees of hydrolysis and varying degrees of antioxidant activities. Hydrolysate generated with crude enzyme extract from sardine (Sardinapilchardus) displayed high antioxidant activity, and the higher DPPH radical-scavenging activity (87 ± 2.1% at 2 mg/ml) was obtained with a degree of hydrolysis of 6%. This hydrolysate was fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P₁–P₈). Fraction P₄, which exhibited the highest DPPH scavenging activity, was then fractionated by reversed-phase high performance liquid chromatography (RP-HPLC). Seven antioxidant peptides were isolated. The molecular masses and amino acids sequences of the purified peptides were determined using ESI-MS and ESI-MS/MS, respectively. Their structures were identified as Leu-His-Tyr, Leu-Ala-Arg-Leu, Gly-Gly-Glu, Gly-Ala-His, Gly-Ala-Trp-Ala, Pro-His-Tyr-Leu and Gly-Ala-Leu-Ala-Ala-His. The first peptide displayed the highest DPPH radical-scavenging activity (63 ± 1.57%; at 150 μg/ml) among these peptides.     

Antioxidant properties of peptide fractions from silver carp (Hypophthalmichthys molitrix) processing by-product protein hydrolysates evaluated by electron spin resonance spectrometry

Silver carp processing by-product protein is usually discarded as an industrial solid waste. In this study the protein was recovered using a pH-shift method, after which seven commercial proteases were separately employed to prepare antioxidative hydrolysates. Among the hydrolysates, pepsin hydrolysates, which had the highest free radical-scavenging activity, were further separated into five peptide fractions, SCPH-I (>10 kDa), SCPH-II (5–10 kDa), SCPH-III (3–5 kDa), SCPH-IV (1–3 kDa), and SCPH-V (<1 kDa), by using ultrafiltration. The antioxidative properties of the peptide fractions were investigated, using a free radical-scavenging assay, by electron spin resonance. The results show that SCPH-V had the highest scavenging effects on DPPH (1,1-diphenyl-2-picrylhydrazyl), hydroxyl and superoxide anion radicals. SCPH-V had potent antioxidant activity in the prevention of the peroxidation of linoleic acid and alleviation of H₂O₂-induced oxidative stress in human intestinal epithelial caco-2 cells. The results indicated that the antioxidant capacity of silver carp by-product hydrolysates could be enhanced by ultrafiltration.     

Bioactivity of hydrolysates obtained from bovine casein using artichoke (Cynara scolymus L.) proteases

The objective of this work was to obtain casein hydrolysates with aspartic proteinases present in extracts from the artichoke flower (Cynara scolymus L.) and evaluate their antioxidant, antimicrobial, and angiotensin-I converting enzyme (ACE) inhibitory activity in vitro. The casein hydrolysates produced by the action of C. scolymus had elevated antihypertensive and antioxidant activity due to their high hydrophobic peptide content (93.84, 96.58, and 90.54% at 2, 4, and 16 h of hydrolysis, respectively). Hydrolysis time and molecular weight (<3 kDa) had a significant influence on the hypertensive and antioxidant activity of the hydrolysates, which were greater at hydrolysis times of 4 and 16 h and corresponding to the <3 kDa fractions. The <3 kDa fraction of the 16 h hydrolysate had an ACE inhibitory activity with a half-maximal inhibitory concentration (IC₅₀) of 71.77 µg peptides per mL; DPPH and ABTS^•+ radical scavenging activities of 6.27 µM and 6.21 mM Trolox equivalents per mg of peptides, respectively; and iron (II) chelation activity with an IC₅₀ of 221.49 µg of peptides per mL. Antimicrobial activity against Enterococcus faecalis was also observed in the hydrolysates. From the peptide sequences identified in the hydrolysates, we detected 22 peptides (from the BIOPEP database) that were already in their bioactive form (AMKPWIQPK, AMKPWIQPKTKVIPYVRYL, ARHPHPHLSFM, DAQSAPLRVY, FFVAPFPEVFGK, GPVRGPFPII, KVLPVPQK, LLYQEPVLGPVRGPFPIIV, MAIPPKKNQDK, NLHLPLPLL, PAAVRSPAQILQ, RELEELNVPGEIVESLSSSEESITR, RPKHPIKHQ, RPKHPIKHQGLPQEVLNENLLRF, SDIPNPIGSENSEK, TPVVVPPFLQP, VENLHLPLPLL, VKEAMAPK, VLNENLLR, VYPFPGPIH, VYQHQKAMKPWIQPKTKVIPYVRY, VYQHQKAMKPWIQPKTKVIPYVRYL) and are reported to display antioxidant, antimicrobial, and ACE inhibitory activity. We also identified 12,116, 14,513, and 25,169 peptide sequences in the hydrolysates at 2, 4, and 16 h, respectively, that were contained in the primary sequence, and these are reported to display ACE inhibitory, antioxidant, dipeptidyl peptidase IV inhibition, antithrombotic, opioid, immunomodulation, antiamnesic, anticancer, chelating, and hemolytic bioactivity.