Hydrolysis Of Fraction Research Articles

Hypocholesterolemic and Antidiabetic Activity of Peptide Fractions Obtained by Ultrafiltration of Protein Hydrolysates From Sunflower (Helianthus annuus) Seeds

Peptide size is a key factor influencing their biological activity. Taking advantage of this relationship, ultrafiltration (UF) emerges as a valuable tool for fractionating complex peptide mixtures obtained from natural sources. Recent studies have focused on the application of this technology to legumes and oilseeds, with the aim of identifying peptide fractions with bioactive potential. The study is aimed at evaluating the in vitro hypocholesterolemic and antidiabetic activity of peptide fractions of protein hydrolysate from sunflower (Helianthus annuus) seeds. The results showed that the highest degree of hydrolysis values was at 120 min with both enzymes: alcalase and flavourzyme with 70.06% and 38.75%, respectively. Protein hydrolysates and peptide fractions showed hypocholesterolemic and antidiabetic activity. The UF membrane process helped to obtain low molecular weight peptide fractions such as the 1–3 kDa fraction that showed greater hypocholesterolemic activity of 79% with both enzymes (alcalase and flavourzyme), while the antidiabetic activity was higher in the peptide fractions < 1 kDa obtained from the hydrolysates with alcalase at 80 min and flavourzyme at 100 min of reaction with a 55.65% and 70.18% inhibition of α‐amylase. The best value inhibition of α‐glucosidase was 34.73% and 46.02% with the peptide fractions < 1 and > 5 kDa obtained from the hydrolysates with alcalase at 20 min and flavourzyme at 100 min. The UF allowed obtaining peptide fractions with different molecular weights with hypocholesterolemic and antidiabetic activity, which could be an alternative for the design and development of nutraceutical products to treatment of chronic degenerative diseases.

Production and fractionation of protein hydrolysate produced from fish sausage (Keropok Lekor) by-products by Lactobacillus casei fermentation: Optimisation and impact of peptide molecular weight on antioxidant and functional properties

Production and fractionation of protein hydrolysate produced from fish sausage (Keropok Lekor) by-products by Lactobacillus casei fermentation: Optimisation and impact of peptide molecular weight on antioxidant and functional properties

Enhanced antihypertensive chicken by-product hydrolysate fraction after its separation by electrodialysis with ultrafiltration membrane (EDUF)

Enhanced antihypertensive chicken by-product hydrolysate fraction after its separation by electrodialysis with ultrafiltration membrane (EDUF)

Impact of Chlorella vulgaris biomass substitution on in vitro bioaccessibility of cookies

This study aimed to produce low-fat cookies (C) by substituting Chlorella vulgaris biomass (0.5% CB1, 1.0% CB2, and 1.5% CB3) and investigating the bioavailability of minerals, total phenolic content, and antioxidant capacities of the cookies. Chlorella sp. microalgae is recognized for its high phenolic content, antioxidant capacity, and as a source of essential minerals. Extractable and hydrolyzable fractions were prepared to determine the total phenolic content and antioxidant capacity. The total phenolic content of CB samples ranged from 200.82 to 274.07 mg GAE/g, with bioaccessibility values from 32.31 to 47.26 mg GAE/g. The CUPRAC method provided the highest antioxidant capacity values (116.57-154.38 µmol TE/g), while the ABTS method showed the highest bioaccessibility values (6.76-9.21 µmol TE/g). Mineral content analysis (Na, Mg, P, K, Ca, Mn, Fe, Cu, Zn, and Se) revealed significant enhancements in the CB samples compared to controls, showing an approximate 2-fold increase in mineral bioaccessibility. Despite extensive research on microalgae-fortified foods, there is a notable gap in knowledge regarding their "in vitro bioaccessibility." This study aims to pioneer the exploration of bioaccessibility and highlight the positive impact of algae-based food consumption on human health.

Optimization of Hydrolysis Conditions and Bio-functional Properties of Whey Protein Hydrolysate Obtained from Ash Gourd (Benincasa hispida) Protease

Background: Enzymatic hydrolysis alters the bio-functionality of whey proteins by generating bioactive peptides. Plant serine proteases as an alternative to animal and microbial proteases, recently considered useful for whey protein hydrolysate preparation to enhance nutritional and bio-functional properties. Objectives: The present study thus aims at optimization of whey protein hydrolyzing conditions employing ash gourd protease (AGP) and evaluation of 2, 2 diphenyl -1-picrylhydrazyl (DPPH) radical scavenging (antioxidant) and angiotensin I- converting enzyme (ACE) inhibition activities and potency of the generated hydrolysate. Methods: Optimization was accomplished using response surface methodology and generating treatment combinations applying central composite design in two steps: first temperature (50-90°C) and pH (6-9) was optimized adding same concentration of AGP (0.3% v/v) at reconstituted WPC (0.6% protein) and then E/S ratio (2-6 %v/v) and hydrolysis time (1-6h) was optimized at reconstituted WPC (4% protein). The effect of hydrolysis time at optimized condition was also evaluated with response to degree of hydrolysis, antioxidant and ACE inhibition activities of the generated hydrolysate and its ultra-centrifuged fractions ≤3kDa and >3kDa. The AGP (Adjusted to activity 2U/mL) obtained after partially purification by three phase partitioning method was used for hydrolysis experiments. Results: The optimum conditions for enzymatic hydrolysis of whey protein to achieve maximum degree of hydrolysis (13.99%) and DPPH radical scavenging activity (23.05%) were observed at 7.48 and 66.40°C. Similarly, the enzyme performed optimally at enzyme by substrate ratio of 5.85% (v/v) for 6 hours of hydrolysis, providing 19.31 % of degree of hydrolysis and 47.71 % of DPPH radical scavenging activity. AGP hydrolyzed whey protein compared to un-hydrolysed whey protein had significantly higher degree of hydrolysis, DPPH radical scavenging and ACE inhibition activity. The lower molecular weight peptides fractions ≤3kDa was found to be more effective (1.45 times) than the peptide fractions >3kDa however was less effective (1.19 times) than the whole hydrolysate regarding antioxidant activity. Similarly, ≤3kDa fraction of hydrolysate had 1.9 times higher ACE inhibition potential as compared to its whole hydrolysate. Conclusion: Overall, the study showed that the AGP employed whey protein hydrolysate has potential for use as a natural antioxidant and ACE inhibition agent.

Trideoxycytidine Diphosphate Promotes Neural Stem Cell Proliferation and Neurogenesis in Mice.

Food-derived nucleic acids exhibit various biological activities and may act as nutrients. Oral ingestion of the nucleic acid fraction (NAF) of salmon milt extract hydrolysates enhances cognitive function in mice, although their active ingredients have not yet been identified, and detailed mechanisms of action are unknown. To identify active ingredients enhancing cognitive function contained in the NAF and its possible underlying mechanism. Because the NAF is rich in trinucleotides, proliferative effects of all 64 types of trideoxyribonucleotides were examined in primary cultured neural stem cells (pcNSCs). The active trideoxyribonucleotide was administered intrahippocampally (5 μg) in 7-wk-old male Institute of Cancer Research (ICR) mice (n = 4-6) or orally (1 mg) 3 times a week for 2 wk in 6-wk-old male ICR mice (n = 5), followed by evaluating neurogenesis and cognitive function by immunohistochemical analysis and spatial recognition test, respectively. The mechanism of action was examined by proteomic analysis of trideoxyribonucleotide-treated pcNSCs using the software DAVID, followed by western blot analysis in pcNSCs and hippocampus. Among all trideoxyribonucleotides, only trideoxycytidine diphosphate (CCC) significantly promoted NSC proliferation (P < 0.05), whereas exposure to putative metabolites of CCC did not affect the proliferation. Intrahippocampal or oral CCC administration in mice increased doublecortin-positive cells in hippocampus (P < 0.05) and enhanced spatial memory (P < 0.05). Proteomic analysis revealed significant alterations in expression of PI3K-Akt signaling-related proteins, including eEF1A2 (P < 0.05). Intrahippocampal CCC administration significantly increased the Akt phosphorylation (p-Akt/Akt) (P < 0.05), which was abolished with the PI3K-Akt inhibitor, LY294002 (P < 0.05). CCC exposure increased p-Akt/Akt (P < 0.05) in pcNSCs, whereas LY294002 or small interfering RNA for eef1a2 suppressed CCC-induced increase in p-Akt/Akt (P < 0.05) and cell proliferation (P < 0.05). A simple food-derived structural molecule CCC promotes NSC proliferation through eEF1A2/PI3K-Akt signaling pathway, thereby enhancing neurogenesis.

POTENTIALITY OF PROTEIN HYDROLYSATE FROM ANADARA GRANOSA AS NUTRACEUTICAL AGENT: ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES

Objective: This study aims to extract protein hydrolysate from Anadara granosa and assess its impact on protein solubility, antioxidant, and antibacterial activities. Methods: Several methods were used, including the isolation of enzyme bromelain, protein extraction from A. granosa, and the breakdown of protein using trypsin and bromelain. Together with its protein solubility, antioxidant activity (IC50) against free radicals (DPPH), the protein hydrolysate's antibacterial activity (MIC and inhibition zone) against Staphylococcus aureus and Escherichia coli was evaluated. Results: With a molecular weight of 10 kDa and an IC50 of 83.81 mg/ml, the trypsin protein hydrolysate fraction showed remarkable antioxidant activity after 5 h of incubation. At a dosage of just 0.25 mg/ml, the A. granosa protein and its corresponding hydrolysate had inhibitory zones against S. aureus and E. coli that were comparable to those observed in samples treated with amoxicillin. Using trypsin as an enzyme for 3 or 5 h produced the strongest hydrolyzed product. The trypsin hydrolysate was better than the bromelain hydrolysate because of its antioxidant and antibacterial activities. Conclusion: Based on the results, antioxidant and antibacterial activities, and protein solubility were influenced by enzymatic hydrolysis.

Moisturizing Effects of Alcalase Hydrolysate Fractions from Haliotis discus Viscera, a Marine Organism, on Human Dermal Fibroblasts, HaCaT Keratinocytes, and Reconstructed Human Skin Tissues.

Haliotis discus, an abalone, is a marine gastropod mollusk that has been cultivated globally owing to its nutritional value and high market demand. However, the visceral parts of H. discus are typically discarded as by-products, highlighting the need to explore their potential value in developing cosmeceuticals and pharmaceuticals. This study investigated the potential moisturizing effects of H. discus visceral tissues. Various hydrolysates from H. discus viscera tissue were evaluated for proximate composition, radical scavenging, and hyaluronidase inhibition activities. Alcalase hydrolysate was isolated using gel filtration chromatography (GFC), and its moisturizing effects were tested on human dermal fibroblasts (HDF), HaCaT keratinocytes, and reconstructed human skin tissue. The Alcalase hydrolysate showed the highest extraction yield, radical scavenging, and hyaluronidase inhibition activities. The Alcalase hydrolysate GFC fraction 1 increased collagen synthesis-related molecules, including procollagen type 1 in HDF and hyaluronic acid-related molecules in HaCaT cells. These moisturizing effects were confirmed in reconstructed human skin tissues by increased levels of aquaporin 3 and filaggrin. Fraction 1 consisted of two main peptides: DNPLLPGPPF and SADNPLLPGPPF. In conclusion, H. discus Alcalase hydrolysate and its fractions have potential moisturizing properties and can be used as cosmeceuticals.

Development and validation of QuEChERS-HPLC method for simultaneous analysis of 5-hydroxymethylfurfural, furfural, and p-cresol in brewery spent grain's hydrolysate

Developing efficient methods to analyze inhibitors resulting from the breakdown of the lignocellulosic matrix in brewery spent grain is crucial for selecting optimal pretreatment strategies and monitoring these compounds during biodigestion processes. However, to date, these determinations have been based on separate methodologies that have not been subjected to the analytical validation stage. This study optimized QuEChERS extraction methods combined with High-Performance Liquid Chromatography (HPLC) analysis to determine 5-hydroxymethylfurfural (5-HMF), furfural, and p-cresol, simultaneously, in the semisolid and liquid fraction of the BSG hydrolysate. The analytical methods were validated by AOAC and INMETRO guidelines. All analytes showed good linearity, with correlation coefficients (R2) greater than 0.97, and low limits of quantification. Recoveries at low, medium, and high levels were 94-116%, repeatability was 0.6-6.8%, and intermediate precision was 0.5-7.6%. The method proved efficient when applied to a set of seven samples. It will contribute to monitoring inhibitory compounds present in biodigestion systems even at low concentrations, which is currently a challenge.

Improved Antioxidant, Antihypertensive, and Antidiabetic Activities and Tailored Emulsion Stability and Foaming Properties of Mixture of Corn Gluten and Soy Protein Hydrolysates Via Enzymatic Processing and Fractionation.

Bioactive peptides and protein hydrolysates have gained considerable attention in the food industry and functional food markets due to their diverse health effects, including antioxidant, antihypertensive, and antidiabetic properties. This study aimed to produce combined soy and corn protein hydrolysates using Alcalase (Al), modification of Al-hydrolysates through sequential hydrolysis using Flavourzyme (Al-FL), cross-linking of Al-hydrolysates using microbial transglutaminase (MTGase) (Al-TG), and fractionation of Al-hydrolysates by ultrafiltration (UF) with molecular weight (MW) cut-off of 100 (Al-F4), 30 (Al-F3), 10 (Al-F2), and 2 kDa (Al-F1). Notably, the < 2 kDa fraction (Al-F1) showcased exceptional biological activities, including antioxidant (81.54% DPPH, 98.02% ABTS), antihypertensive (95.45%), and antidiabetic effects (44.72% α-glucosidase, 77.52% α-amylase), linked to its high hydrophobic amino acid content and low molecular weights (111 and 263 Da). Conversely, the higher molecular weight fraction (Al-TG) excelled in emulsion and foam stability, attributed to its balanced amino acid profile and larger peptides (1385-7057 Da). Our findings reveal that specific protein hydrolysate fractions, particularly Al-F1 and Al-TG, are promising for applications in food and pharmaceutical formulations due to their enhanced biological and functional properties.

Precooked Jack Bean [Canavalia ensiformis (L.) DC] Sprout: Generation of Dipeptidyl Peptidase-IV Inhibitory Peptides during Simulated Digestion.

Bioactive peptides generated from jack bean sprouts are reported to function as dipeptidyl peptidase IV (DPP-IV) inhibitors. However, no studies have investigated the effect of precooking followed by simulated digestion using pepsin-pancreatin to increase DPP-IV inhibitory peptide generation in jack bean sprouts. Therefore, the present study aimed to explore the generation of DPP-IV inhibitory peptides from precooked jack bean [Canavalia ensiformis (L.) DC] sprouts during simulated digestion with pepsin-pancreatin. The results showed that peptide fractions of the sample hydrolysate with molecular weight <1 kDa exhibited the strongest DPP-IV inhibitory activity (84.77%±0.49%) after simulated digestion. This activity was slightly greater than that (74.12%±0.85%) observed prior to simulated digestion. These findings demonstrate that the DPP-IV inhibitory activity of precooked jack bean sprouts can be retained following simulated digestion. Moreover, our investigation revealed the sequences of two novel peptides following simulated digestion with critical amino acids. The presence of alanine and glycine at the penultimate N-terminus of AAGPKP and LGDLLK confirmed the presence of DPP-IV inhibitors. Both peptide sequences are nontoxic and interact with the catalytic sites of enzymes through hydrogen bonds.

Bioactive Properties of Enzymatic Gelatin Hydrolysates Based on In Silico, In Vitro, and In Vivo Studies.

This current study aims to analyze the potential bioactivities possessed by the enzymatic hydrolysates of commercial bovine, porcine, and tilapia gelatins using bioinformatics in combination with in vitro and in vivo studies. The hydrolysate with superior inhibition of angiotensin converting enzyme (ACE) activity was used to treat the D-galactose (DG)-induced amnesic mice. In silico digestion of the gelatins led to the identification of peptide sequences with potential antioxidant, ACE-inhibitory, and anti-amnestic properties. The results of in vitro digestion revealed that the <1 kDa peptide fraction of porcine gelatin hydrolysate obtained after 1 h digestion with papain (PP) (PP1, <1 kDa) potently inhibited ACE, acetylcholinesterase, and prolyl endopeptidase activities at 87.42%, 21.24%, and 48.07%, respectively. Administering the PP1 to DG-induced amnesic mice ameliorated the spatial cognitive impairment and Morris water maze learning abilities. The dentate area morphology in the PP1-treated mice was relatively similar to the control group. In addition, PP1 enhanced the antioxidant capacity in the DG-induced amnesic mice. This study suggests that PP1 could serve as a potential treatment tool against oxidative stress, hypertension, and neurodegenerative diseases.

Angiotensin converting enzyme inhibitory hydrolysate and peptide fractions from chicken skin collagen, as modulators of lipid accumulation in adipocytes 3 T3-L1, after in vitro gastrointestinal digestion

Inhibitory activity against angiotensin-converting enzyme (IAACE) by chicken skin collagen hydrolysate (CSCH) and their peptide fractions before and after in-vitro gastrointestinal digestion, were evaluated; as well as their ability to modulate lipid accumulation in 3T3-L1 adipocytes. Before digestion, peptide fraction <1kDa (F4) showed the highest IAACE (p<0.05) followed by CSCH. After these samples were digested, F4 presented an IAACE with IC50 similar to its digest (DF4) (188.84 and 220.03μg/mL, respectively), which was 2-fold lower (p<0.05) than IC50 of fraction <1kDa from post-digested hydrolysate (FDH) (388.57μg/mL). Nine peptides were identified as the potential ACE inhibitors in F4 and DF4. Addition of DF4 (800μg/mL) reduced(p<0.05) lipid accumulation by 83% within preadipocytes. A 45-60% reduction of lipid accumulation within differentiated adipocytes was obtained by adding FDH and DF4 (regardless the concentration). These results, digested CSCH and F4 with IAACE may be considered as potential adjuvants for obesity treatment.

Bioactive Peptides from Meretrix lusoria Enzymatic Hydrolysate as a Potential Treatment for Obesity in db/db Mice.

Obesity is acknowledged as a significant risk factor for cardiovascular disease, often accompanied by increased inflammation and diabetes. Bioactive peptides derived from marine animal proteins show promise as safe and effective anti-obesity agents by regulating adipocyte differentiation through the AMPK signaling pathway. Therefore, this study aims to investigate the anti-obesity and anti-diabetic effects of bioactive compounds derived from a Meretrix lusoria Protamex enzymatic hydrolysate (MLP) fraction (≤1 kDa) through a 6-week treatment (150 mg/kg or 300 mg/kg, administered once daily) in leptin receptor-deficient db/db mice. The MLP treatment significantly decreased the body weight, serum total cholesterol, triglycerides, and LDL-cholesterol levels while also exhibiting a beneficial effect on hepatic and serum marker parameters in db/db mice. A histological analysis revealed a reduction in hepatic steatosis and epididymal fat following MLP treatment. Furthermore, poor glucose tolerance was improved, and hepatic antioxidant enzyme activities were elevated in MLP-treated mice compared to db/db control mice. Western blot analysis showed an increased expression of the AMPK protein after MLP treatment. In addition, the expression of lipogenic genes decreased in db/db mice. These findings indicate that bioactive peptides, which are known to regulate blood glucose levels, lipid metabolism, and adipogenesis, could be beneficial functional food additives and pharmaceuticals.

Anticancer Activities of Bromelain Hydrolysate of Soy Protein Against Breast Cancer Cells MCF-7

Soybeans contain proteins that have the potential to produce anticancer bioactive peptides. This study aims to determine the anticancer activity of soy protein hydrolysate against MCF-7 breast cancer cells. Soybean protein hydrolyzed by bromelain enzyme 0.5% (w/v) at the optimum temperature and pH for protein hydrolysis using the Bergmeyer and Grassl method. The degree of hydrolysis value of protein hydrolysate was determined by the Alder-Niesen method and the protein profile was analyzed by SDS-PAGE. The hydrolysate with the best degree of hydrolysis value was analyzed for anticancer activity against MCF-7 breast cancer cells by the Presto Blue assay method, and fractionation of protein hydrolysates by gel filtration chromatography (Sephadex G-15). The molecular weight of the peptide was characterized by LCMS/MS. Soy protein hydrolysis using 0.5% (w/v) bromelain enzyme was optimum at 65 ºC and pH 7.0 for 4 hours, with a hydrolysis degree value of 20.57%. The SDS-PAGE analysis showed that the protein hydrolysates had quite thick protein bands in the range of &lt;35 kDa with an IC50 value of 70.37 mg/mL. Based on the LCMS/MS results, the peptide from fractionation has a molecular weight of 5.133 kDa.

Angiotensin-I-converting enzyme and renin inhibitions by antioxidant shrimp shell protein hydrolysate and ultrafiltration peptide fractions

Angiotensin-I-converting enzyme and renin inhibitions by antioxidant shrimp shell protein hydrolysate and ultrafiltration peptide fractions

Phosphomonoesterase and phosphodiesterase activities in the eastern Mediterranean in two contrasting seasonal situations

Abstract. Hydrolysis of dissolved organic phosphorus by marine planktonic microorganisms is a key process in the P cycle, particularly in P-depleted, oligotrophic environments. The present study assessed spatiotemporal variations in phosphomonoesterase (PME) and phosphodiesterase (PDE) activities using concentration kinetics in the eastern Mediterranean Sea in two contrasting situations: the end of winter (including a small bloom period) and autumn. The distribution and regulation of the maximum hydrolysis rate (Vm) and half-saturation constant (Km) of both ectoenzymes were assessed in relation to the vertical structure of the epipelagic layers. PME reached its maximum activities (Vm) after the addition of 1 µM MUF-P (4-methylumbelliferyl phosphate), whereas, for PDE, it was necessary to add up to 50 µM bis(4-methylumbelliferyl)phosphate (bis-MUF-P) to reach saturation state. On average, the Km of PDE was 33 ± 25 times higher than that of PME. The Vm of PME and Vm of PDE were linearly correlated. Conversely to the Km values, Vm values were on the same order of magnitude for both ectoenzymes, with their ratio (Vm PME : Vm PDE) ranging between 0.2 and 6.3. Dissolved organic phosphorus (DOP) and the phosphomonoesterase hydrolysable fraction of DOP explained most of the lack of variability in Vm PME and Vm PDE. On the contrary, Vm of both phosphohydrolase enzymes was inversely correlated to the concentration of dissolved inorganic phosphorus. The particular characteristics of concentration kinetics obtained for PDE (saturation at 50 µM, high Km, high turnover times) are discussed with respect to the possible unequal distribution of PDE and PME among the size continuum of organic material and accessibility of phosphodiesters.

Soluble and hydrolyzable phenolic compounds in date fruits (Phoenix dactylifera L.) by UPLC-QTOF-MS/MS and UPLC-DAD

Soluble and hydrolyzable phenolic compounds in date fruits (Phoenix dactylifera L.) by UPLC-QTOF-MS/MS and UPLC-DAD

First report of antioxidant potential of peptide fraction derived from Colaconema formosanum (Rhodophyta) protein hydrolysates

First report of antioxidant potential of peptide fraction derived from Colaconema formosanum (Rhodophyta) protein hydrolysates

Protein identification and potential bioactive peptides from pumpkin (Cucurbita maxima) seeds.

Pumpkin is an economically important crop all over the world. Approximately, 18%-21% of pumpkins, consisting of peels and seeds by-products, are wasted during processing. In addition, the seeds are rich in protein and have the potency of bioactive peptide production. This study aims to recognize the proteins and investigate the potential bioactive peptides from pumpkin (Cucurbita maxima) seeds. Pumpkin seeds were subjected to hot air drying (HAD) at 55°C for 12 h and freeze-drying (FD) at -80°C for 54 h before they were powdered, analyzed, and precipitated by isoelectric point to obtain pumpkin seed protein isolates (PSPI). PSPI comprised 11S globulin subunit beta, 2S seed storage albumin, and chaperonin CPN60-1. To generate hydrolysate peptides, PSPI was hydrolyzed using papain, pepsin, and bromelain. FD group pepsin hydrolysates had the highest peptide content of 420.83 mg/g. ACE inhibition and DPP-IV inhibition activity were analyzed for each enzymatic hydrolysate. The pepsin hydrolyzed sample exhibited the highest ACE inhibition of 70.26%, and the papain hydrolyzed sample exhibited the highest DPP-IV inhibition of 30.51%. The simulated gastrointestinal digestion (SGID) conducted by pepsin and pancreatin increased ACE inhibitory activity from 76.93% to 78.34%, and DPP-IV inhibited activity increased from 58.62% to 77.13%. Pepsin and papain hydrolysates were fractionated using ultrafiltration to measure ACE and DPP-IV inhibition activity. The highest free radical scavenging abilities were exhibited by the <1 kDa hydrolysate fractions with 78.34% ACE inhibitory activities and 79.55% DPP-IV inhibitory activities. This research revealed that pumpkin seeds had the potency to produce bioactive peptides.