Search Results (268)

Postprandial hyperglycemia represents a critical therapeutic target in type 2 diabetes mellitus (T2DM), requiring interventions that simultaneously address glycemic dysregulation and oxidative stress. This study evaluated the anti-hyperglycemic and antioxidant properties of Sclerocarya birrea leaf crude extract (CE) and biosynthesized silver nanoparticles (AgNPs). Phytochemical screening, nanoparticle characterization (UV–Vis, XRD, TEM, SEM, DLS, FTIR), enzyme inhibition assays (α-amylase, α-glucosidase, DPP-IV), glucose dynamics in Caco-2 cells, and antioxidant assays (DPPH, total antioxidant capacity, H₂O₂ cytoprotection) were performed. Phytochemical analysis identified flavonoids, tannins, alkaloids, and terpenoids as major constituents of Sclerocarya birrea leaf extract. AgNPs exhibited spherical morphology (36.8 ± 8.6 nm, n = 100 particles analyzed), face-centered cubic crystallinity (crystallite size: 32.1 nm), and characteristic surface plasmon resonance at 451 nm. Both formulations inhibited α-amylase (CE IC₅₀: 14 µg/mL; AgNPs IC₅₀: 14.07 µg/mL, n = 3) and α-glucosidase (CE IC₅₀: 15.96 µg/mL; AgNPs IC₅₀: 15.82 µg/mL, n = 3), showing substantial inhibition, though less potent than acarbose. Uniquely, AgNPs demonstrated selective DPP-IV inhibition (IC₅₀: 220.5 µg/mL, n = 3, p < 0.001 vs. CE), completely absent in CE. In antioxidant assays, DPPH scavenging potency was comparable between formulations (CE IC₅₀: 23.45 µg/mL; AgNPs IC₅₀: 22.26 µg/mL, n = 3), while CE achieved higher maximal scavenging at the tested concentrations. Conversely, AgNPs provided superior intracellular cytoprotection against H₂O₂-induced oxidative stress in kidney cells (80.2 ± 2.1% viability at 76 µg/mL vs. CE 69.8 ± 3.4% at 190 µg/mL, n = 3, p < 0.001), representing a 2.5-fold dose advantage. Neither formulation significantly altered glucose uptake or SGLT1 expression in intestinal epithelial cells (p > 0.05, two-way ANOVA, n = 3). These findings establish S. birrea-based formulations, particularly AgNPs, as promising multifunctional candidates for managing postprandial hyperglycemia and oxidative complications in T2DM. They also highlight nanotechnology-enhanced phytomedicine as an innovative therapeutic strategy. Full article

Background/Objectives: Collagen hydrolysates are widely used as nutritional ingredients for skin and joint health; however, growing evidence indicates that collagen may also exert beneficial effects on cardiometabolic pathways. Short peptides have been shown to modulate angiotensin-converting enzyme (ACE) and dipeptidyl peptidase IV (DPP-IV), key regulators of blood pressure and glucose homeostasis. This study aimed to assess the dual ACE- and DPP-IV inhibitory and GLP-1 stimulation activities, respectively of a tripeptide-enriched formulation (CH). The study was performed using a benchmark collagen hydrolysate (BCH) as reference. Methods: ACE and DPP-IV inhibitory activities were evaluated using in vitro enzymatic assays. Cellular compatibility and in situ DPP-IV inhibition were assessed in Caco-2 intestinal cells, while glucagon-like peptide-1 (GLP-1) secretion was measured in STC-1 enteroendocrine cells. The degree of hydrolysis was determined by OPA assay, and nanoLC–HRMS was used to characterize and compare the proteomic profiles of the samples. Results: Both hydrolysates exhibited dose-dependent ACE and DPP-IV inhibition; however, CH showed significantly higher inhibitory activity at comparable concentrations. CH also reduced cellular DPP-IV activity in Caco-2 cells and stimulated GLP-1 secretion in STC-1 cells, whereas BCH showed limited or non-significant cellular effects. Peptidomic analysis revealed an enrichment of short- and medium-length peptides in CH, while BCH contained a higher proportion of long peptides (>2000 Da). Consistently, CH exhibited a 1.7-fold higher degree of hydrolysis than BCH. Conclusions: The tripeptide-enriched collagen hydrolysate demonstrated superior enzymatic and cellular bioactivity compared with the benchmark formulation, supporting its potential as a multifunctional bioactive ingredient for health applications. Full article

Background: Sirtuin 3 (SIRT3), a mitochondrial NAD⁺-dependent deacetylase, plays a central role in regulating mitochondrial metabolism, oxidative stress, and cell survival. Although SIRT3 has been implicated in angiogenesis, apoptosis, and inflammation, its global proteomic impact on the brain remains unclear. This study aimed to systematically characterize alterations in angiogenesis-, apoptosis-, chemokine-, and cytokine-related proteins in the brains of SIRT3 knockout (SIRT3 KO aka SIRT3⁻/⁻) mice compared with wild-type (WT) controls. Methods: Adult male C57BL/6 WT and SIRT3 KO mice were analyzed using proteome profiler antibody microarrays covering 53 angiogenesis factors, 21 apoptosis markers, 28 chemokines, and 111 cytokines. Protein expression changes were quantified by chemiluminescence imaging and densitometric analysis. Results: The results showed a distinct suppression of angiogenic proteins (amphiregulin, angiogenin, DPPIV, GM-CSF, IGFBP-2, IGFBP-3, IL-1β, PDGF-AA, PDGF-BB, proliferin, serpin F1, thrombospeondin-2, TIMP-4, and VEGF-B), activation of both pro-apoptotic (BAD, cytochrome c, Smac/DIABLO, HIF-1α, Fas, TNF R1, and TRAILR2) and anti-apoptotic, stress-related proteins (Bcl-x, catalase, HO/HMOX2, HSP27, HSP70, and MCL1) in the SIRT3 KO animals compared with the WT controls. Notably, SIRT3 deficiency was associated with increased expression of inflammatory mediators linked to glial activation and neurodegeneration (BLC/CCL13, LIX/CXCL5, MIG/CXCL9, chitinase 3-like 1, CCL22/MDC, IL-6, myeloperoxidase, osteopontin, RBP4, Reg3G, and TNF-α), alongside disturbed proteins involved in immune surveillance and vascular remodeling (6Ckine/CCL21, chemerin, DF, EGF, fractalkine/CX3CL1, HGF, IGFBP-6, IL-16, and I-TAC). Conclusions: Collectively, these findings demonstrate that SIRT3 is a key regulator of mitochondrial-dependent vascular, apoptotic, and neuroimmune pathways in the brain, and that its loss creates a molecular environment consistent with heightened vulnerability to neurodegenerative processes. Full article

Background: To battle the side effects of anticancer Pt(II) drug cisplatin, the development of photoactivatable and/or intracellular reduction-activatable Pt(IV) prodrugs has become a promising strategy. Methods: Herein, two novel Pt(IV) prodrugs, namely, cis,cis,trans-[Pt^IV(NH₃)₂(Cl)₂(OH)(probenecid)]) (SPP) and cis,cis,trans-[Pt^IV(NH₃)₂(Cl)₂(probenecid)₂] (DPP) bearing mono- and di-probenecid at the axial positions of oxoplatin have been synthesized via covalently linking of carboxylate group in probenecid, which is a well-established clinic drug by inhibiting organic anion transporter 1 (OAT1) to reduce cisplatin-induced nephrotoxicity, with the axial hydroxyl group(s) in oxoplatin. The promising cytotoxicity of SPP and DPP against MCF-7, T47D breast cancer cells and the MDA-MB-231 triple-negative breast cancer cells was evaluated, and the mechanism of action of the two Pt(IV) prodrugs was investigated by apoptosis assay and Western blot assay. Results: SPP exhibits a comparable cytotoxicity to cisplatin against MCF-7 and T47D breast cancer cells, while it shows 2.1-fold higher cytotoxicity than cisplatin against MDA-MB-231 cells. DPP was shown to be more cytotoxic than SPP, and exhibits 8.7-, 7.5-, and 2.3-fold higher cytotoxicity than cisplatin against MCF-7, T47D, and MDA-MB-231 cells, respectively. Apoptosis assays revealed a similar early-apoptotic cell death mechanism to cisplatin for both SPP and DPP. The enhanced cellular and nuclear uptake of DPP compared to cisplatin contributes to its promising anticancer efficacy. DPP can bind to OAT1 in cancer cells, which may synergistically enhance the cytotoxicity of the Pt(IV) anticancer prodrugs. Conclusions: The direct conjugation of probenecid to the axial positions of oxoplatin confers the resulting Pt(IV) prodrugs a multitargeting property, significantly promoting the cytotoxicity of the resulting Pt(IV) complexes. This finding provides a practical strategy for drug design and cancer treatment based on platinum complexes. Full article

Background/Objectives: This study aimed to evaluate the antidiabetic potential of five extracts/sub-extracts and five known cycloartane saponins [astragalosides (AST) I, II, III, IV, and cyclocanthoside E] from Astragalus noeanus (AN), using four specific diabetes-related molecular targets. Methods: Four diabetes-associated in vitro and in silico targets—protein tyrosine phosphatase 1B (PTP1B), dipeptidyl peptidase IV (DPP IV), α-amylase, and advanced glycation end-products (AGEs)—were employed to obtain comprehensive antidiabetic activity profiles. Additionally, the antioxidant and prebiotic capacities of the extracts/sub-extracts were assessed in vitro. A cycloartane saponin was isolated and structurally characterized. Quantitative analyses of total flavonoids, total saponins, and high-performance thin-layer chromatography (HPTLC) were performed to profile the chemical constituents of the plant material. Results: Among the extracts/sub-extracts, the aqueous extract (ANW) exhibited the highest inhibitory effects against all four diabetes-related targets, with inhibition percentages ranging from 83.70% to 93.49%. The methanol extract (ANM) demonstrated significant prebiotic activity comparable to standard controls on two Lactobacillus strains. The chloroform extract (ANC) showed the highest flavonoid content and exhibited the strongest antioxidant activity across all assays. ANM yielded the highest saponin content (3250 mg escin equivalent/g). HPTLC quantification revealed that AST IV was the predominant saponin in ANM (14.28 μg/mg) after cyclocanthoside E (117.27 ± 6.71 μg/mg). Among the saponins, AST IV displayed the most potent inhibition in diabetes-related enzyme assays, surpassing reference drugs acarbose and vildagliptin at equivalent concentrations. AST III also demonstrated considerable activity, ranking just below AST IV. Molecular docking studies identified AST II and AST III as the most promising ligands, exhibiting superior binding affinities and stronger hydrogen bonding and hydrophobic interactions with target proteins. Cyclocanthoside E was isolated from A. noeanus and evaluated for its antidiabetic effects for the first time, with its structure confirmed by NMR and LC-HRMS analyses. Conclusions: This study highlights Astragalus noeanus as a promising source for safe and effective antidiabetic agents. The potent activity of the aqueous extract, along with AST IV and AST III, warrants further investigation through clinical trials to validate their therapeutic potential in diabetes management. Full article

The objective of this study was to explore the effect of the assembly sequences of wall materials on the structure and properties of Antarctic krill peptide (AKP)-loaded ovalbumin (OVA)–chitosan (CS) nanoparticles (NPs). Two AKP-loaded NPs (CS/OVA-AKP and OVA/CS-AKP) were prepared by changing the sequences of OVA and CS. The results confirmed that CS/OVA-AKP had a smaller particle size (291 nm vs. 320 nm), lower polydispersity index (0.233 vs. 0.282), higher absolute zeta potential (34.4 mV vs. 32.1 mV), and higher encapsulation efficiency (81.6% vs. 75.4%) than OVA/CS-AKP. X-ray diffraction analysis confirmed that AKP was encapsulated in an amorphous state within the NPs. Fourier transform infrared spectroscopy and three-dimensional (3D) fluorescence spectroscopy revealed that electrostatic interactions, hydrogen bonding, and hydrophobic interactions were the primary driving forces for nanoparticle formation, with CS/OVA-AKP demonstrating a stronger OVA fluorescence quenching effect. Compared with OVA/CS-AKP, CS/OVA-AKP exhibited better redispersibility, and CS/OVA-AKP showed greater stability under various environmental factors (thermal treatment, salt concentration, pH, and storage time). During simulated gastrointestinal digestion, CS/OVA-AKP effectively protected AKP from gastric degradation and showed a higher AKP release rate in simulated intestinal fluid (61.1%) than OVA/CS-AKP (53.0%). The release followed the Korsmeyer–Peppas model, with OVA/CS-AKP exhibiting non-Fickian diffusion (n = 0.7500), and CS/OVA-AKP approached Case II transport (n = 0.9889), indicating erosion-controlled release behavior. CS/OVA-AKP also demonstrated higher hypoglycemic activity, with inhibition rates of 41.1%, 37.5%, and 36.1% for α-glucosidase, α-amylase, and DPP-IV, respectively. These findings underscore the important influence of wall-material assembly sequences on the structure and properties of AKP-loaded NPs, offering valuable insights for the development of bioactive peptide delivery systems. Full article

To enhance the screening efficiency of bioactive peptides, an AI-driven approach was employed to screen DPP-IV inhibitory peptides from goat blood proteins by an integrated in silico, in vitro, and machine learning strategy. Furthermore, the inhibitory mechanism of DPP-IV inhibitory peptides was elucidated by kinetics, molecular docking and simulation. Additionally, their in vitro digestive stability was assessed. In silico results revealed that goat blood proteins were promising precursors of DPP-IV inhibitory peptides, while bromelain was the optimal protease. Their peptide sequences were further identified by peptidomics and predicted by self-developed machine learning models (LightGBM) to identify the potent DPP-IV inhibitory peptides. Two novel DPP-IV inhibitory peptides were identified (FPL and FPHFDL). Enzyme kinetics, molecular docking and molecular simulation data indicated that FPL served as a competitive inhibitor, whereas FPHFDL was a non-competitive inhibitor. Overall, the integrative in silico and in vitro strategy is feasible for rapid screening of DPP-IV inhibitory peptides from goat blood proteins. Full article

Background/Objectives: Essential amino acids’ (EAAs) biological effects depend on both gastrointestinal stability and intestinal bioavailability. A commercially available EAA blend has previously shown to be highly bioaccessible and able to inhibit the DPP-IV enzyme both directly and at a cellular level following simulated digestion in vitro. In light with this consideration, the present study aimed to evaluate the intestinal in vitro bioavailability of GAF subjected to INFOGEST digestion (iGAF) and to investigate the metabolic effects of its bioavailable fraction on muscle cells using an integrated Caco-2/C2C12 co-culture model. Methods: Differentiated Caco-2 cell lines were treated with iGAF, and amino acid transport was quantified by ion-exchange chromatography. The basolateral fraction containing bioavailable EAAs was used to treat differentiated C2C12 myotubes for 24 h. Western blot analyses were performed to assess the activation of anabolic and metabolic pathways, including mTOR, Akt, GSK3, AMPK and GLUT-4. Results: More than 50% of each EAA present in iGAF crossed the Caco-2 monolayer, with BCAAs and phenylalanine particularly enriched in the basolateral fraction. Exposure of C2C12 myotubes to the bioavailable iGAF stimulated mTORC1 activation and increased the phosphorylation of Akt and GSK3, indicating an enhanced anabolic response. At a cellular level, iGAF also elevated the p-AMPK/AMPK ratio, suggesting activation of energy-sensing pathways. Moreover, GLUT4 protein levels and glucose uptake were significantly increased. Conclusions: The study focuses exclusively on a cellular model, and results suggested that iGAF is highly bioavailable in vitro and that its absorbed fraction activates key anabolic and metabolic pathways of skeletal muscle cells, enhancing both protein synthesis signaling and glucose utilization in vitro. Full article

During gastrointestinal digestion, dietary proteins are hydrolyzed into peptides and free amino acids that modulate enteroendocrine function and satiety-related hormone secretion along the gut–brain axis, thereby contributing to obesity prevention. We investigated whey protein concentrate (WPC) as a source of bioactive peptides and evaluated the effects of its digests on cholecystokinin (CCK) secretion in STC-1 enteroendocrine cells by integrating the standardized INFOGEST in vitro digestion protocol, peptidomics (LC–MS/MS), and in silico bioactivity prediction. In STC-1 cells, the <3 kDa intestinal peptide fraction exhibited the strongest CCK stimulation, positioning these low-molecular-weight peptides as promising bioactive components for satiety modulation and metabolic health applications. Peptidomic analysis of this fraction identified short sequences derived primarily from β-lactoglobulin (β-La) and α-lactalbumin (α-La), enriched in hydrophobic and aromatic residues, including neuropeptide-like sequences containing the Glu–Asn–Ser–Ala–Glu–Pro–Glu (ENSAEPE) motif of β-La f(108–114). In silico bioactivity profiling with MultiPep predicted antihypertensive, angiotensin-converting enzyme (ACE)–inhibitory, antidiabetic, dipeptidyl peptidase-IV (DPP-IV)–inhibitory, antioxidant, antibacterial, and neuropeptide-like activities. Overall, digestion of WPC released low-molecular-weight peptides and amino acids that enhanced CCK secretion in vitro; these findings support their potential use in nutritional strategies to enhance satiety, modulate appetite and energy intake, and improving cardiometabolic health. Full article

Type 2 diabetes mellitus (T2DM) poses a critical global health burden, necessitating safer multi-target therapies. We pioneer the exploration of novel bioactive peptides from Tenebrio molitor larvae—an underexplored, sustainable, and edible insect protein—through proteomics-guided screening and bioassays. Six unique peptides (DK-7, WK-6, GR-7, FK-8, SK-6, and DK-8) demonstrated significant α-glucosidase and dipeptidyl-peptidase IV (DPP-IV) inhibitory effects, and significant glucose consumption enhancement in insulin-resistant HepG2 cells. Molecular docking revealed a binding topology where peptides interacted with α-glucosidase at its active sites (Glu271, Arg643, Arg647, Arg653, Tyr733, Lys765, and Glu767) and with DPP-IV at active residues (Phe357, Tyr547, Trp629, Asp729, and Gln731) through dual hydrogen-bond networks and hydrophobic interactions, establishing a novel inhibition mechanism. We wish to propose that insect-derived biopeptides have potential value as next-generation therapeutics, simultaneously advancing sustainable drug discovery and approximating functional food bioresources to biomedicine. Full article

This study investigated the potential of scale-up mango leaf extract (MLE) as a treatment for diabetes, a global public health concern. MLE was prepared by boiling in water, yielding 12.07% (w/w), with a bioactive mangiferin content of 165.67 ± 10.88 μg/g in the crude powder. Mechanistically, MLE demonstrated a hypoglycemic effect by stimulating glucagon-like peptide-1 (GLP-1) secretion in NCI-H716 L-cells. This occurred through activation of the MAPK signaling pathway, evidenced by increased p-ERK1/2, p-p38, and p-c-Jun expression, and the Wnt signaling pathway, shown by increased β-catenin and decreased GSK-3β and Axin1 expression, consistent with molecular docking. In a type 2 diabetic rat model, MLE administration (40 mg/kg) significantly reduced metabolic parameters, including fasting blood glucose (FBG), body weight, cholesterol (CHOL), triglycerides (TGs), and HbA1c. Notably, MLE lowered serum insulin and the HOMA-IR index, and reduced serum dipeptidyl peptidase-IV (DPP-IV) levels, resulting in increased serum GLP-1, comparable to the drug sitagliptin. These findings suggest that MLE has great potential to lower blood glucose by inducing GLP-1 secretion via MAPKs and Wnt signaling pathways, positioning it as a promising candidate for alternative diabetes treatment or development as a dietary supplement. Full article

The objective of this work was to evaluate the antidiabetes potential of protein hydrolysates derived from “vaina morada” black bean (Phaseolus vulgaris L.). Bioactive peptide sequences were identified after in silico digestion. The biological activities and molecular interactions of peptides with targeted enzymes were assayed. The degree of hydrolysis and protein profile were evaluated throughout the processing stages, including protein extraction, hydrolysis, and dialysis. Biological potential assays, including antioxidant potential (DPPH and ABTS•+), and inhibition of α-amylase and α-glucosidase enzymes, were performed. Identified bioactive peptides showed potential for inhibiting ACE and DPP-IV, as well as exhibiting antioxidant potential. Molecular docking indicated that several peptide sequences showed equal or stronger binding affinities compared to acarbose. Notably, sequence VNDNGEPTL exhibited binding energies of −10.0 kcal/mol (α-amylase) and −11.8 kcal/mol (α-glucosidase). Protein hydrolysates showed the lowest IC₅₀ (113.16 µM TE/mg for ABTS•+), while dialyzed protein hydrolysates demonstrated the strongest activity for DPPH (IC₂₅ of 38.83 µM TE/mg). Also, the dialyzed hydrolysate demonstrated the highest enzyme inhibition, with IC₅₀ values of 0.78 mg/mL for α-amylase and 0.60 mg/mL for α-glucosidase. “Vaina morada” black bean protein hydrolysates are a rich source of multifunctional peptides, supporting their potential application in functional food formulations aimed at preventing or managing type 2 diabetes. Full article

This study evaluated the nutritional, functional, biological, and sensory potential of proteins derived from Yarrowia lipolytica biomass and their enzymatic hydrolysates for food applications. Three strains were cultivated under bioreactor conditions, with strain JII1c selected for its superior biomass yield and protein content. Its amino acid composition was rich in lysine and branched-chain amino acids, with protein quality indices (CS = 37.8%, EAAI = 36.17%) confirming value in plant-based diets. Proteins were isolated and hydrolysed using a non-commercial serine protease from Cucurbita ficifolia, which enhanced solubility (NSI: 19.4 → 49.2%), water and oil absorption, and emulsion stability. Hydrolysates showed notable biological activities, including ACE (71.8%), DPP-IV (52.3%), and α-glucosidase (67.4%) inhibition, indicating potential metabolic benefits. Sensory evaluation of extrudates confirmed improvements in aroma, texture, and flavour when hydrolysates were incorporated. The use of a plant-derived protease demonstrates a sustainable approach to producing bioactive peptides. Y. lipolytica hydrolysates emerge as promising clean-label ingredients that combine nutritional quality with techno-functional performance, supporting their integration into health-oriented and sustainable food products. Full article

This study aimed to formulate a carrier system to improve the oral bioaccessibility of goat casein peptides (GCAPS). Goat casein was hydrolyzed with papain and subsequently purified to obtain bioactive peptide fractions (GCAPS) with potent hypoglycemic activity. On this basis, spherical GCAPS-loaded nanocarrier systems were developed, including liposomes (GCAPS-LS) and niosomes (GCAPS-NS). Among them, GCAPS-NS exhibited higher encapsulation efficiency (94.98 ± 3.01%) and a smaller particle size (89.81 ± 8.97 nm) than GCAPS-LS. FT-IR analysis confirmed successful peptide encapsulation. Simulated gastrointestinal digestion experiments demonstrated that GCAPS-NS significantly improved GCAPS retention and DPP-IV inhibition. In vivo results from high-fat diet-induced (HFD) insulin-resistant mice demonstrated that GCAPS-NS effectively ameliorated metabolic abnormalities by including adiposity, enhancing GLP-1 levels and suppressing hsCRP expression, thereby contributing to improved glycemic homeostasis. Moreover, GCAPS-NS intervention resulted in a significant enrichment of Akkermansia and a reduced Firmicutes/Bacteroidetes ratio, suggesting its beneficial role in alleviating HFD gut dysbiosis. These findings indicated that goat casein peptides held great potential as a functional food for the management of type 2 diabetes. Full article

The sustainable revalorization of porcine blood is crucial due to the large volumes daily generated in slaughterhouses. The aim of this study was to obtain a novel ingredient rich in free amino acids and bioactive peptides from the sequential hydrolysis of porcine blood. Porcine blood was hydrolyzed with Alcalase 4.0 L and Protana™ Prime enzymes, followed by molecular weight fractionation (<10 kDa) and spray-drying. The antioxidant, hypoglycemic, and anti-inflammatory bioactivities of the resulting hydrolysate (PBSH) were studied in vitro. Further fractionation by reversed-phase high-performance liquid chromatography (RP-HPLC) was performed to isolate the most bioactive fraction based on polarity. Peptides from fraction 1 (F1) were identified using LC-MS/MS and analyzed in silico. Finally, some peptides were synthesized, and their bioactivity was subsequently assessed. PBSH hydrolysate showed antioxidant activity with IC₅₀ values of 2.09, 135.05, and 26.73 mg/mL for ABTS, FRAP, and DPPH assays, respectively. Additionally, PBSH exhibited hypoglycemic, anti-inflammatory, and immunomodulatory potential through the inhibition of DPP-IV (82.78%), NEP (84.72%), TACE (50.79%), and MGL (69.08%) enzymes at a concentration of 20, 20, 100, and 20 mg/mL, respectively. Peptides PDDFNPS, FPPKPKD, DNPIPK, GHLDDLPG, and GDL were identified in the most polar and bioactive fraction (F1) and proved a synergistic hypoglycemic effect at a concentration of 1 mmol/L. The peptide PDDFNPS exhibited multifunctional properties with 56.43% inhibition of DPP-IV and 83.54% inhibition of NEP. PBSH resulted in a novel functional ingredient for animal feed as it contains a variety of identified bioactive peptides and a high amount of free amino acids. Full article