Search Results (206)

The bioactive potential of dairy-derived peptides has attracted increasing interest due to their capacity to exert antioxidant and antihypertensive effects. This study investigated three artisanal cheeses manufactured with animal rennet (CTRL), Onopordum platylepis extract (OP), or a mixture of both coagulants (AR/OP) to compare their peptide profiles and associated bioactivities. Water-soluble extracts were analyzed to identify precursors and released bioactive peptides, and in vitro assays were performed to assess antioxidant activity and angiotensin-converting enzyme (ACE) inhibition. The analysis of precursors suggested a predominance of antioxidant sequences in CTRL and ACE-inhibitory precursors in OP, with AR/OP showing intermediate values. However, direct peptide identification confirmed that the AR/OP mixture produced a wider range of peptides with antioxidant activity, while OP and AR/OP exhibited similarly high levels of ACE-inhibiting peptides. These results were consistent with in vitro assays, which confirmed AR/OP as the most active sample for antioxidant potential and OP, closely followed by AR/OP, as the strongest for ACE inhibitory activity. Overall, the integration of precursor analysis, peptide identification, and experimental validation highlights the influence of the coagulant on the generation of bioactive peptides, suggesting that the use of Onopordum platylepis Murb. (O. platylepis) alone or in combination with animal rennet may enhance the functional properties of cheese. Full article

Valsartan (Val)—a lipophilic non-peptide angiotensin II type 1 receptor antagonist—is highly effective against hypertension and displaying limited solubility in water (3.08 μg/mL), thereby resulting in low oral bioavailability (23%). The limited water solubility of antihypertensive drugs can pose a challenge, particularly for rapid and precise administration. Herein, we synthesize and characterize valsartan-containing silver nanoparticles (Val-AgNPs) using Mangifera indica leaf extracts. The physicochemical, structural, thermal, and pharmacological properties of these nano-conjugates were established through various analytical and structural tools. The spectral shifts in both UV-visible and FTIR analyses indicate a successful interaction between the valsartan molecule and the silver nanoparticles. The resulting nano-conjugates are spherical and within the size range of 30–60 nm as revealed in scanning electron-EDS and atomic force micrographs. The log-normal distribution of valsartan-loaded nanoparticles, with a size range of 30 to 60 nm and a mode of 54 nm, indicates a narrow, monodisperse, and highly uniform particle size distribution. This is a favorable characteristic for drug delivery systems, as it leads to enhanced bioavailability and a consistent performance. Dynamic Light Scattering (DLS) analysis of the Val-AgNPs indicates a polydisperse sample with a tendency toward aggregation, resulting in larger effective sizes in the suspension compared to individual nanoparticles. The accompanying decrease in zeta potential (to −19.5 mV) and conductivity further supports the idea that the surface chemistry and stability of the nanoparticles changed after conjugation. Differential scanning calorimetry (DSC) demonstrated the melting onset of the valsartan component at 113.99 °C. The size-dependent densification of the silver nanoparticles at 286.24 °C correspond to a size range of 40–60 nm, showing a significant melting point depression compared to bulk silver due to nanoscale effects. The shift in Rf for pure valsartan to Val-AgNPs suggests that the interaction with the AgNPs alters the compound’s overall polarity and/or its interaction with the stationary phase, complimented in HPTLC and HPLC analysis. The stability and offloading behavior of Val-AgNPs was observed at pH 6–10 and in 40% and 80% MeOH. In addition, Val-AgNPs did not reveal hemolysis or significant alterations in blood cell indices, confirming the safety of the nano-conjugates for biological application. In conclusion, these findings provide a comprehensive characterization of Val-AgNPs, highlighting their potential for improved drug delivery applications. Full article

Nanoemulsions (NEs) offer a promising strategy for delivering lipophilic cannabidiol (CBD) to protect skin from particulate matter (PM)-induced damage. In this study, CBD-loaded oil-in-water NEs based on Brij^® O10 (polyoxyethylene (10) oleyl ether) and olive oil were prepared by the phase inversion temperature (PIT) method and characterized. A 20% w/w Brij^® O10 formulation (B20) remained clear and stable for 30 days. CBD solubility was markedly enhanced in Brij^® O10 micelles and further increased in NEs, exceeding theoretical predictions and indicating synergistic solubilization in the oil–surfactant system. CBD incorporation lowered the PIT and induced nonlinear changes in droplet size with oil content. All formulations exhibited nanoscale droplets by dynamic light scattering and transmission electron microscopy, moderately low zeta potentials consistent with nonionic steric stabilization, and maintained physical stability despite increased turbidity at higher oil levels. In a full-thickness human ex vivo skin model exposed to PM, both blank and CBD-loaded NEs reduced interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) in PM-exposed skin, with CBD-loaded NEs providing additional reductions and uniquely restoring procollagen type I C-peptide (PIP) relative to their blanks. Overall, PIT-based CBD NEs enhance CBD solubilization and protect human ex vivo skin from PM-induced inflammation and extracellular matrix degradation. Full article

This study aimed to identify novel angiotensin-converting enzyme (ACE)-inhibitory peptides from royal jelly proteins (RJPs) by integrating in silico digestion, virtual screening, and in vitro evaluation. Three major royal jelly proteins (MRJP1-3) were subjected to in silico digestion using 16 enzymatic systems, yielding 1411 unique peptides. Virtual screening based on predicted bioactivity, toxicity, water solubility, and ADMET profiles resulted in the selection of 27 candidate peptides. Molecular docking revealed strong binding affinities for these peptides compared with the positive control captopril, among which PYPDWSFAK and RPYPDWSF exhibited potent ACE-inhibitory activity, with IC₅₀ values of 110 ± 1.02 μmol/L and 204 ± 0.61 μmol/L, respectively. Kinetic analysis indicated that PYPDWSFAK acts as a mixed-type ACE inhibitor. Docking visualization demonstrated that PYPDWSFAK forms multiple hydrogen bonds with key residues in the ACE active pocket and directly coordinates with the catalytic Zn²⁺ ion. Cellular assays showed that PYPDWSFAK was non-cytotoxic, suppressed Ang II–induced endothelial cell migration, restored NO and ET-1 balance, and enhanced SOD and GSH-Px activities. Overall, this study enriches the repertoire of ACE-inhibitory peptides derived from royal jelly proteins. Furthermore, PYPDWSFAK is identified as a promising ACE-inhibitory peptide with potential for incorporation into natural antihypertensive ingredients or functional foods. Full article

Umami peptides were screened and identified from yak bone collagen for the first time by in silico analysis, molecular docking, and electronic tongue. Twenty proteases with known cleavage sites were used for the simulated proteolysis, and results indicated that “pepsin + papain” was the optimal enzymatic strategy for yak bone collagen to generate peptides with potential umami taste. Moreover, 82 novel unreported peptides with umami taste were found from the simulated hydrolysate, among which 9 peptides exhibited high binding affinity with the T1R1/T1R3 receptor (both -CDOCKER energy and CDOCKER interaction energy > 40 kcal/mol) via molecular docking. Subsequently, six novel umami peptides were identified through sensory evaluation and electronic tongue analysis, including VY, VM, SL, SN, VN, and IS (umami sensory score > 5). These peptides were also in silico characterized with high hydrophobicity, good water solubility, non-toxicity, non-allergenicity, good intestinal absorption, and good oral bioavailability. Furthermore, the identified peptides could bind with the key residues (such as HIS281 and LEU304) within the Venus flytrap domain of the T1R3 subunit of receptor T1R1/T1R3 through hydrogen bonds and electrostatic attractions to generate umami perception. This study revealed the mechanism of umami peptides identified from yak bone collagen and provides a novel approach for the development of umami peptides from animal sources. Full article

The presence of antioxidants in food contributes to the preservation of its taste and technological qualities, preventing its spoilage for a longer time, which is important at all stages of production and storage. The major antioxidants are vitamins, proteins (primarily, enzymes), peptides, amino acids, fatty acid residues of lipids, etc. There is currently an explosive growth in the development of methods for assessing the content and effectiveness of particular antioxidants but not the total antioxidant activity (AOA) in raw milk and food systems. This article provides a critical overview of the most important AOA methods, their mechanisms and applicability, advantages, and limitations (primarily, for antioxidants of milk and dairy products). Among all the antioxidant indicators of milk, the simplest and sufficiently informative is the detection of the total amount of water-soluble antioxidant (TAWSA), which is confirmed by comparison of numerous publications and practical results of various methods (as summarized in this review). It is important to emphasize that the TAWSA of milk is an “integral characteristic” of the most valuable biosubstances (possessing AOA) together. Therefore, the TAWSA method is recommended for assessing AOA in raw milk as an “integrated indicator” in dairy husbandry. Full article

Peptides are notable cosmetic ingredients owing to their diverse biological activities and beneficial effects on skin health. Therefore, multifunctional peptides capable of simultaneously exerting antioxidant, whitening, and anti-wrinkle effects are highly desirable. In this study, a scalable and cost-effective chemical synthesis strategy was used for the rapid design and synthesis of linear peptide sequences with skin bioactivity using solid-phase peptide synthesis. Subsequently, liquid-phase peptide synthesis was used to enhance the proteolytic stability and develop a cyclic peptide, cyclic CYGSR (CR5), which was subjected to in vitro biological evaluation. CR5 showed high biocompatibility in water-soluble tetrazolium salt-1 (WST-1) assays, maintaining over 90% cell viability at concentrations up to 400 μg/mL. In the 2,2-Diphenyl-1-picrylhydrazy (DPPH) assay, CR5 exhibited strong antioxidant activity with 83.18% radical scavenging at 200 μg/mL. It also showed 97.79% tyrosinase inhibition at 800 μg/mL, confirming significant whitening potential. Moreover, CR5 inhibited matrix metalloproteinase-1 (MMP-1) expression by 73.55% and increased type I procollagen expression by 44.68% at 400 μg/mL, demonstrating its anti-wrinkle potential. Additionally, molecular docking and dynamic simulation demonstrated stable binding of the peptide to tyrosinase and MMP-1. Collectively, CR5 possesses multifunctional properties with excellent biocompatibility, highlighting its potential as a novel cosmetic active ingredient. 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

A growing scientific interest in bioactive compounds from sea cucumbers is contributing to a broader recognition even in regions where their consumption is not common. This study evaluated the biological potential of a Holothuria forskali extract obtained through different extraction methods, including water extraction, ethanol–water extraction, and enzymatic hydrolysis. The hydrolysate (H), rich in low-molecular-weight peptides, yielded the highest antioxidant (30.6 ± 0.6 mg VitC Eq/g sample for ABTS and 10.7 ± 0.1 mg GAEs/g sample for Folin-reactive substances) and ACE-inhibitory (82.6%) activities. Based on these results, the hydrolysate was selected for encapsulation in two nanostructured delivery systems for comparative purposes: chitosan nanoparticles (NPs) and rapeseed lecithin liposomes (LPs). Both nanostructures were characterized in terms of size, ζ-potential, and polydispersity and subjected to simulated in vitro gastrointestinal digestion (GIDv) to assess their stability and mucoadhesive properties. After digestion, antioxidant activity increased in both systems, particularly in liposomes. Although encapsulation initially reduced ACE-inhibitory activity, gastrointestinal digestion restored or enhanced it, especially in liposomal formulations (≈37% inhibition). The mucoadhesive potential of the nanostructures after DGIv, focusing on their interactions with mucin, was assessed. Liposomal digests significantly increased viscosity in the presence of mucin, while chitosan nanoparticles decreased it, suggesting the formation of soluble complexes with reduced hydrodynamic volume. Electrostatic and hydrogen bonding interactions between chitosan and mucin were particularly evident in the NPH formulation. The rheological synergism parameter (Δη) revealed more negative values for NPs and NPHs, indicating stronger mucoadhesive interactions compared to controls and suggesting their suitability for mucosal delivery. These findings support the use of H. forskali hydrolysates as a source of functional bioactive compounds and highlight the potential of chitosan-based nanocarriers for enhancing their stability, bioaccessibility, and mucoadhesive properties in functional food or nutraceutical applications. Full article

While the starfish species Asterias pectinifera (Ap) and Asterias amurensis (Aa) are considered ecological threats to marine environments and the fishing industry, recent studies have identified them as rich sources of highly water-soluble, non-toxic collagen peptides. Mitochondrial dysfunction is a key driver of cellular senescence and skin aging, yet the therapeutic potential of marine-derived extracts in modulating mitophagy remains largely unexplored. In this study, we investigated whether starfish-derived extracts could mitigate senescence-associated phenotypes in human dermal fibroblasts (HDFs) through the modulation of mitophagy. Treatment with Ap- or Aa-derived extracts led to reduced senescence-associated β-galactosidase (SA-β-gal) activity, decreased expression of matrix metalloproteinase-1 (MMP-1), and suppression of pro-inflammatory cytokines including interleukin-6 (IL-6) and interleukin-8 (IL-8). Ap- or Aa-derived extracts significantly increased mitophagy in HDFs stably expressing mitochondrial-targeted Keima (HDF-mtKeima), while knockdown of PINK1, the essential regulator of mitophagy, abolished the mitophagy-inducing effects of Ap- or Aa-treatment, indicating that Ap- or Aa-derived extracts activate PINK1/Parkin-dependent mitophagy pathways. Importantly, PINK1 knockdown reversed starfish-induced suppression of MMP-1 and p21, demonstrating its crucial role in regulating senescence-associated gene expression. Additionally, Ap or Aa treatments significantly reduced reactive oxygen species (ROS) accumulation, improved mitochondrial function, and enhanced both basal and maximal respiratory capacity in senescent HDFs. These findings highlight that extracts derived from starfish promote mitophagy through PINK1-dependent mechanisms, exhibiting significant anti-senescence effects in HDFs. This suggests their potential application in the development of novel cosmeceuticals with skin-protective and rejuvenating properties. Full article

Background/Objectives: Nanocrystals (NCs) are a relatively underexplored yet adaptable platform with broad potential for various applications. Currently, the surface modification of NCs leads to the development of versatile platforms capable of enhancing targeted delivery potential and supporting the advancement of precision medicine. With this in mind, this study focused on the design and surface functionalization of a resveratrol (RSV) NC selected for its antioxidant and neuroprotective effects. Methods: The design of the RSV NC was assessed by the Quality by Design approach. With the aim of intranasal administration, we assessed the RSV NC functionalization with a cationic poly (amino acid) belonging to the class of cell-penetrating peptides. Both naked and surface-modified RSV nanosuspensions were characterized in terms of mucoadhesion, behavior in artificial cerebrospinal fluid, crystallinity, solubility, and storage stability. The scavenging activity (%) of neat RSV and its nanosized forms was measured using the DPPH assay. Results: RSV NCs were successfully designed, producing truncated cubic crystals (~240 nm) with an ~80% drug content. Functionalization was efficiently achieved with poly-l-arginine hydrochloride as revealed by DSC and FTIR and resulted in a positively charged nanosuspension. Nanonization technology improved drug solubility in water and did not affect RSV scavenging activity. Technological characterization demonstrated that both nanosuspensions present suitable properties for intranasal administration in terms of particle size, mucoadhesive tendency, and stability in artificial cerebrospinal fluid. An MTT assay revealed the safety of all treatments in human microglia (HMC3) cells. Conclusions: RSV NCs’ functionalization enhanced their brain delivery potential, establishing a promising platform to improve therapeutic outcomes in neurodegenerative diseases. Full article

Biologically active peptides can be obtained with various research methods, depending on the starting material, biological activity, and intended use. To use the most efficient method, it is worth combining in silico and in vitro experiments. Among the tools that can support an in silico analysis are databases such as the Antimicrobial Peptide Database (AMPD) or BIOPEP-UWM. The aim of this study was to make an in silico hydrolysis of peptides with anticancer properties selected from the AMP database, using pepsin, trypsin, and chymotrypsin. Most peptides obtained had properties inhibiting ACE and dipeptidyl peptidase IV activity. Among the resulting peptides, those with the sequence AR, CF, ER, TF, IY, ER, AW, GF, TW, SK and IM are potentially resistant to peptidase from microbial action. An analysis of the peptides’ characteristics showed that peptides with the sequence AR, EK, ER and SK are well-soluble in water and have high affinity for protein and ligand binding. Peptides with the sequence TF, IL and PF are unstable. Thermostable peptides are PGL, IL, GL, IY, VF, PL, IM and QL. The results of the study may be used to design in vitro experiments. Full article

The aim of this study was to design a poorly water-soluble electrostatic nanocomplex of semaglutide (SMG) with protamine sulfate (PS) and zinc ions (Zn) for prolonged subcutaneous delivery. Complexation of SMG with the cationic peptide PS increased the lipophilicity (logP) proportionally from −4.7 to 0.3, particularly in the presence of Zn. The optimized nanocomplex exhibited spherical morphology, an amorphous state, a particle size of 196.0 nm, and a zeta potential of −45.7 mV. In an in vitro dissolution test under sink conditions, native SMG showed rapid drug release with 98% dissolution within 24 h. In contrast, the nanocomplexes showed markedly delayed release, with a concentration-dependent relationship between PS/Zn contents and SMG release rate, exhibiting 19% drug release over 7 days in the optimized formula. These findings suggest that the proposed nanocomplex is a promising system for long-acting injectable delivery of SMG, potentially enhancing patient compliance in patients with obesity or type 2 diabetes. Full article

The growing population and increasing concerns about food security and sustainability demand innovative solutions to minimize food waste and transform by-products into functional ingredients valuable to the food sector. Brewery by-products, including brewer’s spent grain (BSG) and brewer’s spent yeast (BSY), are underutilized resources despite their high protein contents and potential as sustainable food ingredients. This study aimed to transform BSG and BSY into protein hydrolysates (BSGH and BSYH, respectively) through enzymatic hydrolysis and thus add value to these brewery industry by-products to be used in the food industry. These protein hydrolysates were incorporated into non-alcoholic malt beverages at three different concentrations, and their effects on the physicochemical properties, including color, kinematic viscosity, turbidity, foaming capacity and foam stability, of the non-alcoholic malt beverages were evaluated. Both BSGH and BSYH exhibited higher water solubility (WS) and lower water binding capacity (WBC) values when compared to their native non-hydrolyzed forms, enhancing their suitability as ideal ingredients for protein supplementation of a wide range of food and beverage products. The production of peptides of varying sizes underscored the effectiveness of enzymatic hydrolysis which resulted in an increase in cysteine and methionine levels in BSYH but a decrease in BSGH. The addition of BSGH and BSYH increased the kinematic viscosity and turbidity but reduced the lightness values in color of the non-alcoholic malt beverages. When the properties of the protein hydrolysates were compared, BSYH was more effective than BSGH in forming foams and maintaining their stability for longer periods. These findings highlight the potential of brewery by-products, after enzymatic hydrolysis, as protein-rich ingredients that can support more sustainable food systems and contribute to the nutritional enhancement of various low-protein food and beverage products. Full article

The global transition toward plant-based diets has intensified the search for sustainable protein alternatives, positioning hemp-based meat analogs (HBMAs) as a promising solution due to their exceptional nutritional profile and environmental benefits. This comprehensive review critically examines hemp protein research, focusing on extraction technologies, nutritional excellence, functional innovation, and sustainable processing approaches for meat analog development. Hemp seeds contain 25–30% protein, primarily consisting of highly digestible edestin and albumin proteins that provide a complete amino acid profile comparable to soy and animal proteins. The protein exhibits superior digestibility (>88%) and generates bioactive peptides with demonstrated antioxidant, antihypertensive, and anti-inflammatory properties, offering significant health benefits beyond basic nutrition. Comparative analysis reveals that while alkaline extraction-isoelectric precipitation remains the industrial standard due to cost-effectiveness ($2.50–3.20 kg⁻¹), enzymatic extraction and ultrasound-assisted methods deliver superior functional properties despite higher costs. Hemp protein demonstrates moderate solubility and good emulsifying properties, though its gelation capacity requires optimization through enzymatic hydrolysis, high-pressure processing, or strategic blending with complementary proteins. Processing innovations, particularly high-moisture extrusion combined with protein blending strategies, enable fibrous structures closely mimicking conventional meat texture. Hemp protein can replace up to 60% of soy protein in high-moisture meat analogs, with formulations incorporating wheat gluten or chickpea protein showing superior textural attributes. Despite advantages in nutritional density, sustainability, and functional versatility, HBMAs face challenges including sensory limitations, regulatory barriers, and production scaling requirements. Hemp cultivation demonstrates 40–50% lower carbon footprint and water usage compared with conventional protein sources. Future research directions emphasize techniques and action processes, developing novel protein modification techniques, and addressing consumer acceptance through improved sensory properties for successful market adoption. Full article