Cowpea Protein Research Articles

Enhanced characterization and utilization of cowpea protein isolate: rheological and textural properties of heat-induced gels for plant-based egg omelet formulation

This study aimed to evaluate the potential of cowpea isolate (CPI) as an effective plant-based egg substitute by comparing its composition and physicochemical properties with those of commercial isolates, including mung bean, soy, and pea proteins. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis identified a distinctive vicilin-like globulin in CPI that is critical for the unique functional properties of CPI. CPI has a smaller particle size, resulting in superior gelation capacity and water retention, which are essential for egg substitute applications. Thermal analysis revealed the high stability of CPI, which is advantageous for food processing. Notably, the unique heat-induced gelation behavior of CPI, characterized by a smooth transition from solid- to liquid-like states, provided a strength not observed in its commercial counterparts. Microstructural analysis showed that the CPI gel closely resembled the egg gel, underscoring the potential of CPI to mimic the texture of eggs, particularly in omelets.

In Vitro Digestion and Fermentation of Cowpea Pod Extracts and Proteins Loaded in Ca(II)-Alginate Hydrogels.

Antioxidants derived from food by-products are known for their bioactive properties and impact on human health. However, the gastrointestinal behavior is often poor due to their degradation during digestion. The development of Ca(II)-alginate beads supplemented with biopolymers and enriched with cowpea (Vigna unguiculata) extract could represent a novel environmentally friendly technological solution to produce functional ingredients in the food industry. The present study evaluates the impact of in vitro digestion/fermentation by analyzing global antioxidant response (GAR), production of short-chain fatty acids (SCFAs) as a modulation of gut microbiota, and behavior of proton transverse relaxation times by low-field nuclear magnetic resonance (as an indicator of gelation state and characterization of microstructure). Results revealed that guar gum and cowpea protein preserved a high GAR of total phenolic compounds and antioxidant capacity by ABTS and FRAP methods after digestion/fermentation, promoting an adequate protection of the bioactives for their absorption. Alginate-based beads have great potential as prebiotics, with the guar gum-containing system contributing the most to SCFAs production. Finally, the overall higher mobility of protons observed in the intestinal phase agrees with structural changes that promote the release of phenolic compounds during this stage. Beads are excellent carriers of bioactive compounds (cowpea phenolic compounds and peptides) with potential capacities.

Antithrombotic and anticariogenic activity of peptide fractions from cowpea (Vigna unguiculata) protein hydrolysates.

Protein-derived peptide fractions can play a key role in the physiological and metabolic regulation and modulation of the body, which suggests that they could be used as functional ingredients to improve health and to reduce the risk of disease. This work aimed to evaluate the in vitro antithrombotic and anticariogenic bioactivity of hydrolysates and protein fractions obtained from cowpea (Vigna unguiculata) by biocatalysis. Cowpea protein concentrate was hydrolyzed by sequential action with two enzyme systems, Pepsin-Pancreatin or Alcalase-Flavourzyme. There was extensive enzymatic hydrolysis, with degrees of hydrolysis of 34.94% and 81.43% for Pepsin-Pancreatin and Alcalase-Flavourzyme, respectively. The degree of hydrolysis for the control treatments, without the addition of the enzymes Pepsin-Pancreatin and Alcalase-Flavourzyme was 1.1% and 1.2%, respectively. The hydrolysates were subjected to fractionation by ultrafiltration, with five cut-off points according to molecular weight (<1, 1-3, 3-5, 5-10 and >10 kDa). The Alcalase-Flavourzyme hydrolysate led to 100% inhibition of platelet aggregation, while the Pepsin-Pancreatin hydrolysate showed 77.41% inhibition, but this was approximately 100% in the ultrafiltered fractions. The highest anticariogenic activity was obtained with the Pepsin-Pancreatin system, with 61.55% and 56.07% for calcium and phosphorus demineralization, respectively. Hydrolysates and their peptide fractions from Vigna unguiculata exhibited inhibition of platelet aggregation and protection of tooth enamel and have the potential for use in the development of functional products with beneficial health effects. © 2024 Society of Chemical Industry.

Exploring the rheological and thermal behavior of cowpea protein concentrate: Impact of pH and concentration

This study confronts challenges from global population growth, focusing on protein supply strains and the environmental impact of traditional animal sources. Delving into unexplored territory, this study investigates the combined effects of pH, concentration, and temperature on the rheological and thermal properties of cowpea protein concentrate (CPC). Steady-state and oscillatory rheological tests, along with differential scanning calorimetric analysis, were conducted to unveil the behavior of CPC solutions. The results revealed sensitivity to protein concentration and shear rate, especially at higher concentrations (>10%), and pH emerged as a critical factor, significantly influencing shear stress and viscosity. The elastic and viscous components of alkaline CPC solutions exhibit weak gel behavior, influenced by both pH and frequency. Temperature-induced changes in viscosity and gelation highlight the interaction between thermal conditions and protein structures. The thermal cycle helps explain changes in molecular interactions induced by temperature, with pH playing a role in influencing gelatinization and denaturation. The denaturation temperature (Td) in the thermograms corresponds to vicilin fractions and emphasizes the prominence of denaturation and gelation at neutral pH, where Td was pH 7 > pH 9 > pH 11. Alkaline pH induced significant structural changes, resulting in a gelation onset temperature below the denaturation onset temperature. This study provides a deeper understanding into the rheological and thermal properties of CPC solutions, emphasizing the roles of concentration, pH, and temperature, useful for various steps of industrial processing, before and after thermal gelation. These findings are important for promoting the use of cowpea proteins in the food industry in response to the evolving needs of a growing global population.

Potential use of cowpea protein hydrolysate as a dietary supplement in broiler chickens: effects on growth, intestinal morphology, muscle lipid profile, and immune status

In the current study, we tested the effects of dietary cowpea protein hydrolysate (CPH) on broilers’ growth, carcase traits, intestinal histomorphology, The fatty acid profile of breast muscle, blood biochemical parameters, and immune status. Three-day age male Ross-308 broilers (n = 300, average weight 86.72 g ± 0.20) were distributed into five groups and fed on five diets supplied with five levels of CPH at 0, 2, 4, 6, and 8 g kg−1 for 35 days. The results revealed that dietary CPH did not change (P < 0.05) the broilers’ growth performance or carcase traits, except for a quadratic decrease (P < 0.05) in FCR at concentrations 2 and 4 g kg−1 during the finisher period. The villus heights (VH) and goblet cell count (GCC) were increased (P < 0.05) in the duodenum and jejunum; however, the villus width decreased (P < 0.05) in the duodenum without showing changes (P < 0.05) in the jejunum and ileum. Dietary supplementation with 6 g CPH kg−1 increased (P < 0.05) the duodenal crypt depth (CD). Dietary supplementation with 8 g CPH kg −1 increased the VH: CD in the duodenum compared to the control; nevertheless, a quadratic (P < 0.05) change was reported in the jejunum. The VH: CD did not change (P < 0.05) in the ileum; however, dietary supplementation with 6 CPH kg−1 increased (P < 0.05) the ileal VH and GCC. The inclusion of CPH led to a linear (P < 0.05) increase in the total n-3 polyunsaturated fatty acids (PUFA), n-6 PUFA, and n-3/n-6 PUFA in broilers’ breast muscle. An increase in the assessed metabolic and immune status indices was reported, as evidenced by increases (P < 0.05) in thyroid hormones, growth hormone, total protein, albumin, globulin, lysozymes, interferon-gamma, interleukin10, and complement 3 serum levels. However, the serum glucose and leptin levels did not show significant (p > 0.05) changes in response to dietary CPH. Moreover, the dietary CPH levels 4–8 g kg−1 upregulated the immunostaining of IgG in the spleen of these groups. These findings suggested that including CPH in broiler diets could be an effective candidate for modulating the chickens’ metabolic and immune status; however, it had no increased effect on broiler growth.

Amino acid composition and angiotensin-i converting enzyme inhibitory activity of Cowpea (&lt;i&gt;Vigna unguiculata&lt;/i&gt;) pepsin-pancreatin protein hydrolysate fractions

Legumes are high in protein and have been linked to the management and prevention of chronic diseases due to their bioactive components. This study investigated the amino acid composition of cowpea protein hydrolysate and its inhibitory impact on angiotensin-I converting enzyme (ACE-I). Cowpea (Vigna unguiculata) protein hydrolysate (CPH) was produced through sequential digestion with pepsin and pancreatin. CPH was then separated using G-50 gel filtration chromatography and reverse phase high performance liquid chromatography (RP-HPLC). The fractions' ACE-I inhibitory activity was then assessed. The IC50 value for the RP-HPLC fraction with the highest ACE-I inhibitory activity (P&lt;0.05) was 6.23 μg/mL. The amino acid profile revealed that a large proportion of hydrophobic amino acids were present in the RP-HPLC fraction containing the peptide with the highest ACE-I inhibitory activity. These findings suggest that CPH has the potential to be employed in the development of functional foods that can aid in the prevention and treatment of hypertension.

Effect of pH on Foaming Properties (FC and FS) of Protein Isolates from Two Varieties (DAS &amp; BS) of Nigerian Cultivated Solojo Cowpea (Vigna Unguiculata L. Walp)

Effect of pH on the foaming properties (FC and FS) of protein isolates from two Varieties (DAS &amp; BS) of Nigerian cultivated Solojo cowpea (Vigna Unguiculata L. Walp) was investigated. Results show that the foaming capacity (FC) followed the solubility trend, with a high foaming capacity at pH 2, which slowly reduced between pH 4 - 6 and gradual increase again with pH as the pH approached 10, where it had another high value. As shown in Figures 1-6 noticeable changes in FC were detected as the pH of the protein solution progressed from pH 2 to 10, with minimum foaming capacity at pH 4 in all cases except for DFDAS flour and BS protein isolate (pH 6). The foaming capacity value for 6 h soaked and 48 hr germinated flours, at pH 4 were observed to be greater than at pH 2 and 6. While the maximum was observed in some cases at pH 8 and some pH 10 but FFDAS Raw, 36 and 48 hr; FFBS, 6 hr; DFBS, Raw and 72 hr; BS, 6 and 24 hr germinated, all had their own max values at pH 2. Defatting was also observed to improve the foaming capacity values of all the treatment. The foaming capacity ranged between 15.19 to 121.16%; 44.76 to 127.36%; 31.23 to 110.93%; 30.84 to 121.16%; 64.97 to 210.00%; 62.67 to 222.00% for FFDAS, DFDAS, FFBS, DFBS, DAS, and BS, respectively. The pattern of FC change followed the pH dependent solubility flow of the test material, showing the importance of solubility on foam formation which confirms that solubility of protein has a great influence on the activities of foam also increasing the pH from 4 to 10 improved the foaming characteristics of flour. This observation proved the authenticity that foaming property depends on protein solubility. The foaming capacity of cowpea protein isolates was pH dependent and high at alkaline pH in comparison to acid region.

Modifications to functional and biological properties of proteins of cowpea pulse crop by ultrasound-assisted extraction

Natural resource depletion, negative environmental effects and the challenge to secure global food security led to the establishment of the Sustainable Development Goals (SDGs). In need to explore underutilized sustainable protein sources, this study aims at isolating protein from cowpea by ultrasound-assisted extraction (UAE), where the techno-functional characteristics of the protein isolates were studied at different sonication conditions i.e., 100 W and 200 W at processing times ranging from 5 to 20 min. The US at 200 W-10 min produced the optimal results for all properties. In this process combination, there was an increase in protein yield, solubility, water-holding capacity, foaming capacity and stability, emulsion activity and stability, zeta-potential, and in-vitro protein digestibility from 31.78% to 58.96%, 57.26% to 68.85%, 3.06 g/g to 3.68 g/g 70.64% to 83.74%, 30.76% to 60.01%, 47.48% to 64.26%, 56.59% to 87.71%, –32.9 mV to −44.2 mV and 88.27% to 89.99%, respectively and particle size dropped from 763 nm to 559 nm in comparison to control. The microstructure and secondary-structure alterations of proteins caused by sonication were validated by SEM images, SDS-PAGE, and FTIR analyses. Sonication leads to acoustic cavitation and penetrate the cell walls, improving extraction from the solid to liquid phase. After sonication, the hydrophobic protein groups were exposed and proteins were partially denatured which increased its functionality. The findings demonstrated that UAE of cowpea protein improved yield, modify characteristics to fit the needs of the food industry, and contribute to achieving SDGs 2, 3, 7, 12, and 13.

Emulsifying Capacity of Cowpea Protein Isolates. Effect of Thermal and Hydrolytic Treatment.

In this work, modifications due to the effect of thermal treatments (TT 70 and 90°C) and partial hydrolysis by digestion with alcalase (LH) on the emulsifying properties of cowpea protein isolates (CPIs) extracted at pH 8 and 10 were analyzed. In addition, the influence of protein concentration [0.1 and 1% (w/v)] was evaluated. Emulsions (O:W) were prepared and particle size, stability, interfacial composition, and microstructure were studied. Fresh emulsions formulated with TT CPIs presented lower values of volume-weighted mean droplet size (D4.3), with the increase in temperature and treatment time, compared to the untreated CPIs. After seven days of storage, D4.3 and the indexes of flocculation (FI) and coalescence (CI) increased, mainly at 90°C. On the other hand, the emulsions with LH CPIs presented lower D4.3 values compared to all the conditions tested, remaining unchanged during the storage time. The destabilization process in the TT CPIs emulsions revealed coalescence at 0.1% (w/v) and cremated-flocculation at 1% (w/v). The presence of polypeptides of low molecular mass (MM) at the interface would be responsible for the better stability found in emulsions with LH CPIs, compared to those formulated with untreated and TT CPIs. Increasing the protein concentration resulted in a significant improvement of all emulsifying properties.

Optimizatoin of Anti-Nuntritional Removal, Protein Extraction and Modification of Functional Properties of Cowpea (Vigna unguiculata L.) Protein with Sonication

Cowpea has high protein content which makes it is potential to be utilized as a protein isolate. However, cowpeas contain some anti-nutritional compounds which can lower the nutritional quality of the cowpeas. It is necessary to reduce the antinutritional contents from the cowpeas.The objective of this study was to investigate the optimum treatments for antinutritional removal and protein extraction to produce protein isolate. Removal of antinutritional was performed by a combination of soaking treatment with the duration of 24, 48, and 72 hours with 5% NaHCO3 solution and boiling for 0, 2.5, and 5 minutes. The protein extraction was conducted by adjusting the pH to various pH at 2, 4, 6, 8, 10, 11, and 12.Soaking of cowpeas seeds for 48 hours with 5% NaHCO3 significantly reduced HCN levels to 0.72 ppm, phytic acid of 9.62 mg / g, and trypsin inhibitor of 7.02 mg / g. The protein concentrate was obtained from protein extraction by using optimum pH at 12 while precipitation of protein by using pH 4 with protein concentration at 70.9%. Sonication with 80%-30 minutes indicated an increase in the water holding capacity, oil holding capacity, emulsion stability, foaming capacity, and stability rather than untreated

Prediction and Evaluation of Bioactive Properties of Cowpea Protein Hydrolysates

Cowpea protein hydrolysates were prepared using thermolysin, alcalase, and trypsin and analysed for bioactive properties, and then, the release of bioactive peptides was investigated in silico. It was found that the degree of hydrolysis reached 48% after 24 h hydrolysis with alcalase. The hydrolysate prepared using alcalase showed higher ACE inhibitory (62%) and DPPH scavenging activity (19%). SDS-PAGE analysis revealed that vignin was the major protein in cowpea protein isolate. In silico analysis indicated the presence of potential bioactive peptides with potent bioactivity in the primary structure of proteins. The 3D structure of proteins was built, upon which bioactive peptides were mapped using their location in the primary structure. The secondary structure and solvent accessible surface around each bioactive peptide were then calculated. On this basis, the higher degree of hydrolysis and bioactive properties of cowpea protein hydrolysate prepared by alcalase were explained, and structural factors influencing the release of bioactive peptides were investigated.

Physicochemical and nutritional characteristics of nutrition bar fortified with cowpea protein

Physicochemical and nutritional characteristics of nutrition bar fortified with cowpea protein

Allergenic risk assessment of cowpea and its cross-reactivity with pea and peanut.

Novel protein sources can represent a risk for allergic consumers. The aim of this study was to evaluate the allergenicity of cowpea (Vigna unguiculata), an increasingly consumed legume and potential new industrial food ingredient which may put legume-allergic patients at risk. Children with allergy to legumes associated to peanut (LP group: n= 13) or without peanut allergy (L group: n= 14) were recruited and sensitization to several legumes including cowpea was assessed by prick tests and detection of specific IgE (sIgE). Cowpea protein extract was analyzed by SDS-PAGE and immunoblotting, IgE-reactive spots were subjected to mass spectrometry. IgE-cross-reactivity between cowpea, pea, and peanut was determined using ELISA inhibition assays. Basophil activation tests were performed to evaluate sensitivity and reactivity of patient basophils toward legumes. Prick tests and sIgE levels to cowpea were positive in 8/14 and 4/13 patients of the L group and in 9/13 and 10/13 patients of the LP group, respectively. Four major IgE-binding proteins were identified as vicilins and seed albumin. Cowpea extract and its vicilin fraction strongly inhibited IgE-binding to pea and peanut extract. Peanut, lentil, and pea were the strongest activators of basophils, followed by cowpea, soybean, mung bean, and lupin. A majority of patients with legume allergy were sensitized to cowpea proteins. Four novel allergens were identified in cowpea, among which storage proteins were playing an important role in IgE-cross-reactivity, exposing legume-allergic patients to the risk of clinical cross-reactivity to cowpea and thus adding cowpea to the group of nonpriority legumes that are not subjected to allergen labeling such as chickpea, pea, and lentil.

Comparison of grain protein profiles of Brazilian cowpea (Vigna unguiculata) cultivars based on principal component analysis

This study aims to compare the grain protein profile of four Brazilian cowpea cultivars (BRS Aracê, BRS Itaim, BRS Pajeú, and BRS Xiquexique) by two-dimensional electrophoresis (2-DE) and principal component analysis (PCA). 2-DE efficiently separate cowpea protein profiles, showing high homogeneity among the four cultivars. In addition, the principal component analysis indicated that there is a difference in abundance of proteins among the cultivars. The cultivars BRS Aracê and BRS Xiquexique, both biofortified in iron and zinc, were separated from the cultivars BRS Itaim and BRS Pajeú. These results demonstrate that protein profiles can be used to discriminate cowpea varieties.Graphical

Leveraging Indian pulses for plant‐based meat: functional properties and development of meatball analogues

SummaryWorldwide, the development of next‐generation plant protein‐based meat analogues has gained popularity because of plant proteins' reliable, sustainable, functional and nutritional benefits. In this work, we study the functional properties of green gram (GG), horse gram (HG) and cowpea protein concentrates (CPPCs) obtained using the alkaline/isoelectric precipitation method and developed the conventional and plant‐based deep‐fried meatballs. The results indicated that all protein concentrates exhibited superior functional properties. All protein concentrates exhibited a typical U‐shaped solubility curve from pH 2 to 9. Emulsification, foaming and gelling properties were found to correlate well with the protein solubility (PS) profile. The least gelation capacity for GGPC was 15%, while HGPC and CPPC required 20% protein to form a gel. Water absorption capacity (WAC), emulsification properties were significantly different for all protein concentrates, but not oil absorption, foaming and protein solubility. A plant‐based deep‐fried meatball formulated using the ratio of 20:20:20 for GGPC: HGPC: CPPC was found to be closely related to the conventional meatballs in terms of taste, flavour and colour. The oil absorbed in plant‐based deep‐fried meatballs was ~4% lower than in conventional deep‐fried meatballs. This research allowed a more in‐depth understanding of the functional characterisation of pulse protein concentrates (PPCs) obtained by alkaline‐isoelectric precipitation and their potential use in developing plant protein‐based meatball analogues. In conclusion, Indian pulses could potentially offer sustainable options for plant protein‐based meat formulations.

Legume protein concentrates from green gram, cowpea, and horse gram

In recent years, owing to the rise in awareness regarding sustainability challenges associated with meat production, the trend of plant protein-based meat substitutes as the sustainable and healthier alternative is growing. The present work focuses on the extraction of plant proteins from Indian pulses as a first step towards contributing to this trend. Herein, experiments were designed to develop processes for securing pulse protein concentrates from green gram, horse gram, and cowpea via wet methods, namely alkaline extraction, acid extraction precipitation, water extraction precipitation, and micellization precipitation. The proximate composition of the pulses was determined, the protein yield, protein recovery, and protein purity were estimated to assess the suitability of the wet processing methods. The results showed that the alkaline extraction exhibited the highest yield ranging from 13.5% to 18%, the highest protein recovery varied from 58% to 67%, and the highest purity (cowpea protein concentrate ~79%, horse gram protein concentrate ~79%, and green gram protein concentrate ~87%) compared to other methods. The extraction method significantly influenced the yield and recovery, but not the purity. These results indicated that alkaline extraction is preferable for the purification of proteins from the selected pulses. Novelty impact statement The protein solubility is more, and co-precipitation of non-protein components was significantly less at pH 9. Alkaline extraction exhibited the highest yield of protein, recovery, and protein purity. Alkaline extraction is preferable for the purification of proteins from the pulses.

Optimation 1,2 formulation of meat analog from cowpea (Vigna unguiculata L Walp) protein curds and cocoyams (Xanthosoma sagittifolium) modification starch as filler

Meat analog is a plant protein product that comes from non-meat materials. The materials were used in this research were cowpea curd protein (20−30%) and cocoyam modification starch (20−30%) by autoclaving-cooling. This research aimed to ascertain the optimum formulation of cowpea curd protein as a raw material and cocoyam modification starch as a filler in the meat analog. The data obtained were processed with response surface methodology (RSM) to find the optimum formulation. The optimum results of the central composite design (CCD) showed that the moisture response had a 2FI (2-factor interaction) model with a final equation of Y=52.05−1.73X1+1.16X2+1.84X1X2, where X1X2 Curd protein*modified starch. The ash content had a 2FI model with a final equation of Y=2.79−0.71X1+0.43X2−1.02X1X2, the fat content had a linear model with a final equation of Y=2.59+0.11X1+1.26X2, the protein content had a linear model with a final equation of Y=18.26+0.62X1+0.056X2, the carbohydrate content had a linear model with a final equation of Y=24.54+1.17X1−2.38X2, the texture value had a mean model with a final equation of Y=0.072, and the color value had a mean model with a final equation of Y=30.41. The optimum condition of cowpea curd protein was added by 30% and cocoyam modification starch was added by 30% a with response of water content 50.92%, ash content 2.11%, protein content 18.97%, fat content 4.18%, carbohydrate content 23.82%, texture 0.075 (mm/gr/dt), and color 22.44 (L).

High hydrostatic pressure- or heat-induced gelation of cowpea proteins at low protein content: Effect of calcium concentration

The effect of different calcium levels (10–40 mM) during protein denaturation on the characteristics of cowpea protein gels obtained by heating or high hydrostatic pressure treatment (HHPT) at low protein content (7.5%w/w) was studied.Calcium allowed the formation of self-standing gels. For the lowest levels of treatments (70 °C and 400 MPa) calcium avoided a complete protein denaturation, which in turn resulted in weak matrices. For the highest levels (90 °C and 600 MPa) the complete protein denaturation in the presence of calcium allowed that this ion strengthened the matrices by establishing associations between exposed reactive sites. For HHPT, hardness and elastic module increased for each increase in calcium concentration, while for heating a plateau was reached from 15 mM. Differences in interactions that stabilized aggregates (more hydrophobic ones for heating and more calcium bridges for pressurizing) could explain the differences between heat- and HHPT-induced gels. Most of the gels belonged to the aggregate type, which entails opacity and low water holding capacity.

Indigestible cowpea proteins reduced plasma cholesterol after long-term oral administration to Sprague-Dawley rats

Cowpea protein isolate (CPI) was subjected to various dry and wet heat pretreatments followed by sequential digestion with pepsin and pancreatin; the undigested residues were isolated as the indigestible cowpea proteins (ICPs). All the ICPs exhibited in vitro bile acid-binding capacity but ICP from the slow cooling-induced gelation had the highest yield (68%) and was used for rat feeding experiments to determine effect on plasma total cholesterol (TC). Groups consisting of 3 male and 3 female Sprague-Dawley rats each were fed hypercholesterolemic diets that contained casein only or casein that was partially substituted with ICP of CPI for 6 weeks. Results showed diet that contained 5% (w/w) ICP was more effective in preventing TC increase (1.8 mmol/L) when compared to increases of 9.34 and 4.15 mmol/L for CPI and casein only diets, respectively.Graphical abstract

Structural and antioxidant properties of cowpea protein hydrolysates

Cowpea protein isolates extracted at pH 8.0 (I8) and pH 10.0 (I10) were hydrolysed by alcalase to obtain limited (I8LH and I10LH) and extensive (I8HH and I10HH) hydrolysates. Hydrolysis degree was 2.3% for I8LH, 4.6% for I10LH, 25.8% for I8HH and 24.1% for I10HH. The effect of hydrolysis was assessed by using solubility-pH profile assay, exclusion chromatography, gel electrophoresis, infrared and fluorescence spectroscopy and surface hydrophobicity. Subsequently, ABTS.+, ORAC and HORAC methods were applied to measure antioxidant activity in all samples. Electrophoresis, chromatography and surface hydrophobicity showed different profiles for I8LH and I10LH, presumably, due to the initial extraction pH of protein isolates. Limited hydrolysis increased antioxidant activity in both I8LH and I10LH with values higher than 70% for ABTS.+, 60% for ORAC and 45% for HORAC when compared to their respective isolates. Concerning extensive hydrolysis, I8HH and I10HH yielded similar effects when considering physicochemical, functional and antioxidant properties. Both I8HH and I10HH showed increments above 80% for ABTS.+, 80% for ORAC and 70% for HORAC in comparison to their respective isolates. These results indicate that limited and extensive alcalase hydrolysis of both isolates increases antioxidant activity regardless of the pH for initial protein extraction. Finally, peptides with a molecular weight between 1.8 and 6.5 kDa (I8HH and I10HH from FPLC soluble fractions) exhibited the highest ORAC activity. Thus, cowpea alcalase hydrolysates emerge as viable ingredients to formulate foods with antioxidant properties.