Search Results (308)

In this study, nine different Propionibacterium freudenreichii strains were isolated from Kars Gravyer produced by traditional methods in Turkey and identified by sequencing the 16S–23S intergenic region using species-specific primers. The isolated strains were examined in vitro for the presence of the β-galactosidase enzyme, autoaggregation ability, sensitivity against eight selected antibiotics and survivability under harsh conditions in order to determine their potential probiotic properties. After probiotic potentials were evaluated, an experimental design was made to optimize the production of vitamin B12 in a 3 L glass bioreactor P. freudenreichii NUV774. While all strains showed similar resistance (92–98%) to gastric juice (0.3% pepsin, pH 3.0), they showed resistance to intestinal fluid (0.1% pancreatin, 0.3% bile salt, pH 8.0) between 60% and 92%. It was determined that the viability after 3 and 6 h of incubation in 0.5% and 1% bile salt differed between strains. All isolates exhibited resistance to ciprofloxacin, ampicillin, and trimethoprim–sulphamethoxazole; however, most were sensitive to ofloxacin. Overall, P. freudenreichii strains showed resistance to the gastrointestinal tract, tolerance to pH 3.0, and high tolerance to bile salts. As a result of optimization, maximum vitamin B12 production was found to be 156.8 mg/L. The optimum operating conditions were calculated as temperature = 36.9 °C, aeration = 2.430 vvm, and agitation = 159.120 rpm. Hence, P. freudenreichii, as future probiotic strain candidates, will offer an alternative source to Lactobacillus, Bifidobacterium and some Bacillus spp. In addition, this study denoted that the alteration of the production of active vitamin B12 by P. freudenreichii occurs in a strain-dependent manner. Full article

Spirulina (Arthrospira platensis) is recognized as a high-protein microalga with potential for bioactive peptide production. In this study, S. platensis protein extract (~45% protein) was subjected to enzymatic hydrolysis using pepsin, trypsin, and chymotrypsin. A ~75% reduction in Bradford values indicated extensive protein breakdown, with degrees of hydrolysis of 15.6%, 21.4%, and 33.7% for pepsin-, trypsin-, and chymotrypsin-treated samples, respectively. SDS-PAGE confirmed the generation of low-molecular-weight peptides (<10 kDa). Hydrolysis caused only minor changes in amino acid composition, maintaining protein quality, with trypsin-hydrolysates showing the highest protein efficiency ratio (1.12) and biological value (78.83%). Antioxidant capacity increased significantly, with hydrolysates displaying a 33–68% rise in DPPH and 30–54% in FRAP activity, alongside a 33–44% reduction in lipid peroxidation. Furthermore, phytochemical content was markedly enhanced in hydrolysates compared to intact protein, with increases in total phenolic content (38–118%), total flavonoid content (59–78%), and terpenoids (24–37%). Among treatments, trypsin-SPPH (Spirulina platensis protein hydrolysate) consistently exhibited the most pronounced improvements. Collectively, these findings demonstrate that proteolysis of S. platensis proteins not only enhances antioxidant activity but also liberates bound phytochemicals, establishing S. platensis hydrolysates as promising functional food and nutraceutical ingredients. Full article

This study evaluated the in vitro antioxidant and antiproliferative activity of hemp seed (Cannabis sativa L.) protein isolate, protein hydrolysate, and its fractions. The protein hydrolysate was obtained through sequential enzymatic digestion using pepsin and pancreatin, achieving a degree of hydrolysis of 48.11%. The hydrolysate was then fractionated by ultrafiltration. Assays conducted on Caco-2 (colorectal cancer) and THP-1 (leukemia) cell lines revealed that the higher-molecular-weight fraction of (>10 kDa) exhibited the strongest, concentration-dependent antiproliferative effect, as determined by the neutral red uptake (NRU) assay for Caco-2 cells and the MTT assay for THP-1 cells. Furthermore, a significant intracellular antioxidant activity was observed, particularly in the whole hydrolysate and its low-molecular-weight fractions, as measured by the DCFH-DA assay in Caco-2 cells. The results suggest the potential application of hemp seed protein hydrolysate and its fractions as antioxidant and chemoprotective supplements in oncologic therapies. Full article

The excessive use of synthetic preservatives poses significant threats to food safety and human health. This study systematically investigated the genetic characteristics of the Limosilactobacillus fermentum (L. fermentum) z-6 strain, the antibacterial properties of the bacteriocin-like substance (FC) it produces, and its mechanism of action. The results demonstrated that this bacteriocin-like substance exhibited remarkable antimicrobial activity and exceptional stability, maintaining high activity across a broad pH range (4.0–8.0) and withstanding heat treatment at 100 °C and UV irradiation, indicating robust environmental adaptability. Its proteinaceous nature was confirmed by its detection below 1 kDa on Tricine-SDS-PAGE and its inactivation by trypsin and pepsin. The FC showed broad-spectrum inhibition against foodborne pathogens, including Salmonella enterica serovar Typhimurium, E. coli, A. baumannii, S. aureus, P. mirabilis and L. monocytogenes. Mechanistic investigation demonstrated that the FC exerts antibacterial effects primarily through membrane disruption, as evidenced by a live-dead staining assay confirming significantly enhanced permeability in Salmonella enterica serovar Typhimurium, and scanning electron microscopy revealing distinct pore formation on bacterial surfaces. It is speculated that the FC produced by z-6, due to its excellent properties and outstanding antibacterial performance, could potentially serve as a natural biopreservative. Full article

Background: Chronic gastritis (CG) involves gastric mucosal imbalance, with H. pylori (>90% cases), acid-pepsin imbalance, and bile reflux as druggable mechanisms. FDA-approved drugs show limited efficacy against antibiotic-resistant strains and fail to target undruggable pathways (e.g., inflammation, autoimmune atrophy). Traditional Chinese Medicine (TCM), particularly Qing-Wei-Zhi-Tong micro-pills (QWZT), offers multi-target advantages, though its mechanisms remain poorly understood. Methods: The dual-algorithm integration framework predicts QWZT’s pharmacological effects to treat gastritis. For druggable processes (pathways targeted by existing drugs), the structure–target–pathway similarity algorithm quantifies QWZT similar activities to FDA drugs, validated by gastrointestinal smooth muscle experiments. For undruggable processes (novel biological mechanisms not addressed by current therapies), the multi-target perturbation algorithm predicts QWZT’s unique capacity to undruggable processes and is validated via LPS-induced inflammation in RAW264.7 and GES-1 cells. Results: Structure–target–pathway similarity algorithm identified QWZT compounds sharing prokinetic mechanisms with FDA drugs, validated by dopamine-induced relaxations and acetylcholine-induced contractions in gastrointestinal smooth muscle. Multi-target perturbation algorithm quantified QWZT’s superior disruption of undruggable immune/inflammation networks, confirmed by restored cell viability in LPS-injured GES-1 cells and significantly reduced the expression of NO, IL-6, and TNF-α in RAW264.7 cells via key compounds (paeoniflorin and berberine). Conclusions: QWZT may exert its regulatory effects on gastrointestinal smooth muscle by mediating muscarinic and dopamine receptor D2 (DRD2), and reduce the expression of NO, IL-6, and TNF-α to achieve anti-inflammatory effects, thereby effectively treating CG. The integration strategy that integrates algorithms and experiments to reveal the common and distinct mechanisms of QWZT compared to FDA-approved drugs, offering a novel approach for studying Traditional Chinese Medicine mechanisms. Full article

The increasing consumption of fish products has resulted in significant waste generation, with sardine scales (Sardina pilchardus) representing a notable by-product of the canning industry. This work investigates the sustainable valorization of these scales through collagen extraction for applications in cosmetics and nutrition. Collagen was extracted using acid and pepsin-assisted methods and characterized by spectroscopic and imaging techniques (UV–Vis, FTIR, PXRD, SEM, and SDS-PAGE). Subsequent enzymatic hydrolysis with papain produced low-molecular-weight peptides. Biological assays revealed enhanced antioxidant activity of the hydrolyzed peptides compared to native collagen, while no antimicrobial effects were detected. Permeation studies in Caco-2 cells indicated moderate intestinal absorption (~6.4% in 6 h). These findings support the potential of sardine-derived collagen as a bioactive ingredient aligned with circular economy principles. Full article

The growing demand for functional foods has stimulated the search for novel microbial strains with probiotic potential, such as Veillonella atypica ATCC 17744, which has been emerging as a promising strain. Therefore, the present study aimed to perform an in vitro characterization of this strain, focusing on safety aspects and functional properties such as stress tolerance (pH, bile salts, and simulated gastrointestinal conditions), adhesion capacity (hydrophobicity, auto-aggregation, and biofilm formation), anti-pathogenic activity, antioxidant activity, antibiotic susceptibility, and enzymatic synthesis ability (gelatinase, lipase, catalase, and hemolytic activity). Stress tolerance assays revealed that this strain is sensitive to pH values below 4.00; however, no reduction in cell viability was observed at pH 3.00 in the presence of pepsin or 0.3% and 0.6% bile salts. Hydrophobicity testing showed moderate tolerance to toluene and low tolerance to xylene. Regarding biofilm synthesis, this strain formed a weak biofilm after 48 h of incubation. No anti-pathogenic activity was observed against Streptococcus aureus or Escherichia coli, and it exhibited low antioxidant activity in the DPPH assay. Regarding its safety properties, this strain was sensitive to all tested antibiotics and did not synthesize gelatinase, lipase, catalase, or exhibit β-hemolytic activity. Therefore, Veillonella atypica ATCC 17744 presents promising characteristics supporting its potential application in the development of functional food formulations, although further studies are required to ensure its safety for human consumption. Full article

The study presents the potential of milkfish frame, a by-product of milkfish processing, as a source of dipeptidyl peptidase IV (DPP-IV) inhibitory and antioxidant peptides with potential applications in type 2 diabetes management. Proteomic analysis identified key proteins, including 65 kDa warm temperature acclimation protein 1 and myosin heavy chain. In silico prediction (BIOPEP-UWM) guided the selection of proteases for generating DPP-IV inhibitory peptides. Enzymatic hydrolysates were produced and evaluated for bioactivity. Among the treatments, pepsin hydrolysis (2% v/v, 8 h) yielded the highest peptide content (283.64 mg/g), soluble protein (86.46%), and DPP-IV inhibitory activity (68.47%). The resulting milkfish frame pepsin hydrolysate (MFH) was further enhanced through ultrafiltration and simulated gastrointestinal digestion, which improved the DPP-IV inhibitory and antioxidant capacities. Cytotoxicity assays confirmed that MFH (0–100 μg/mL) was non-toxic to FL83B hepatocytes after 24 h. Moreover, treating TNF-α-induced FL83B cells with 10 μg/mL MFHs improved cell viability, reducing the toxicity induced by TNF-α in cells. These findings show that MFHs exhibit promising antidiabetic potential and could serve as natural alternatives to synthetic drugs for type 2 diabetes management. This also demonstrates the valorization of fish processing by-products into functional food ingredients, advancing sustainable approaches in food innovation. Full article

This work evaluates the effect of raw material and protease enzymes on the antioxidant and antidiabetic potential of fish by-product hydrolysates. For this, mackerel (Scomber scombrus) and sardine (Sardina pilchardus) by-products were hydrolyzed using papain, pepsin, and Protamex^TM. Pepsine produced hydrolysates with a lower degree of hydrolysis (34%) and longer peptide chain lengths (2.9), regardless of the raw material. The highest DH was found for the sardine by-products hydrolyzed with papain and Protamex^TM, exceeding 55% for both enzymes. The mackerel by-product hydrolysates exhibited higher antioxidant activity, while the sardine samples showed more potent antidiabetic effects. Accordingly, sardine by-products and pepsin would be preferable for producing hydrolysates with antidiabetic potential, and mackerel by-products, hydrolyzed papain, and Protamex^TM would be useful for producing antioxidant peptides. This study demonstrates the potential of Atlantic mackerel and sardine waste as a source of bioactive peptides and the opportunity for revalorizing these by-products. Full article

The tree bean (Parkia timoriana), an underutilized legume valued for its nutritional profile, represents a potential source of bioactive peptides for diabetes management. To our knowledge, this is the first study to identify and characterize DPP-IV inhibitory peptides derived from tree bean seed protein hydrolysates. The tree bean proteins were digested with trypsin, thermolysin, chymotrypsin, pepsin, and simulated gastrointestinal (SGI) enzymes, among which SGI hydrolysis yielded the highest degree of hydrolysis (14%) and strongest DPP-IV inhibitory activity (IC₅₀ = 1289 ± 58 µg/mL). Guided by DPP-IV inhibitory assays, sequential fractionation using strong cation exchange and RP-HPLC yielded the most potent fraction, H5, with an IC₅₀ of 949 ± 50 µg/mL. After peptide identification and synthesis, APLGPF (AF6) emerged as the most potent inhibitor, with an IC₅₀ of 396 ± 18 µM. Enzyme kinetics revealed a non-competitive inhibition mechanism, corroborated by molecular docking, which indicated binding at an allosteric site of DPP-IV. Furthermore, AF6 remained stable under simulated gastrointestinal digestion and enzymatic exposure, highlighting its resistance to proteolysis. Taken together, these findings highlight P. timoriana as an underexplored source of peptides with DPP-IV inhibitory activity and identify AF6 as a promising lead for developing functional foods or nutraceuticals aimed at type 2 diabetes management. 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

Thanks to its tolerance to drought, sorghum is a cereal crop that is extensively cultivated in the sub-Saharan region. Its good nutritional value makes it an interesting raw material for the food industry, although several anti-nutritional features pose a challenge to exploiting its full potential. In this study, we evaluated whether the process of germination may represent a way of improving the macro- and micro-molecular profile of sorghum, lowering the content of anti-nutritional factors, and promoting the synthesis of bioactive compounds. Germination for 48 and especially 72 h promoted the hydrolysis of starch and proteins, enhanced antioxidant activity, increased the content of polyphenols, mainly flavonols and flavanones, and promoted the conversion of γ- to α-isomers of tocopherols. At the same time, it significantly reduced the concentration of phytates and linoleic acid, enhancing pepsin activity and contributing to the inaugural examination of the impact of sprouted sorghum on digestive protease activity. These findings could help to promote the utilization of sprouted sorghum as a premium ingredient for food products, providing significant nutritional advantages. Full article

Bacteriocins are ribosomal synthesized antimicrobial peptides produced by bacteria, but their low yields limit industrial applications as food preservatives. This study aimed to optimize the culture conditions of Pediococcus acidilactici CCFM18 and investigate the biological properties of the bacteriocin. The culture temperature, initial pH, and culture time significantly affected the growth of P. acidilactici CCFM18 and bacteriocin production. The optimal culture conditions determined through response surface methodology (RSM) were a culture temperature of 35 °C, an initial pH of 7.0, and a growth time of 16 h. Under these conditions, bacteriocin production reached 1454.61 AU/mL, representing a 1.8-fold increase compared to pre-optimization levels. Biological characterization revealed that the bacteriocin exhibited strong thermal stability (up to 100 °C for 30 min) and pH stability (pH 2–9), but was sensitive to proteolytic enzymes, including pepsin, trypsin, papain, and protease K. The bacteriocin demonstrated antimicrobial activity against both Gram-positive and Gram-negative bacteria, including Enterococcus faecalis and Escherichia coli. These findings provide a theoretical basis for the industrial production and application of the bacteriocin. Full article

This study aimed to develop innovative functional gummy candies enriched with protein hydrolysates derived from porcine liver, enhancing their antioxidant and antimicrobial properties. First, the overall consumer acceptability (OA) was assessed to determine the most suitable combination of gummy matrix components. Selected combinations were then analyzed for antioxidant activity (ABTS•+, DPPH•), antimicrobial effects, microbiological safety, and physicochemical characteristics. The incorporation of liver hydrolysates significantly increased antioxidant capacity. The highest activity was observed in sample GC5Pa24Ag, hydrolyzed with papain for 24 h and formulated with agar, showing ABTS•+ and DPPH• scavenging activities of (67.6 ± 0.98 µmol/g) and (49.14 ± 1.00%), respectively (p ≤ 0.05). Pepsin hydrolyzed samples (GC2Pe3Gl, GC2Pe6Gl, GC2Pe24Gl) exhibited significantly larger inhibition zones against Listeria monocytogenes ATCC 13932, Escherichia coli ATCC 25922, and Salmonella enterica subsp. enterica serovar Typhimurium ATCC 14028 compared to the control (p < 0.05). Among all, GC5Pa24Ag demonstrated the broadest antimicrobial activity, with a 29.0 ± 0.2 mm inhibition zone against all tested pathogens. These findings suggest that porcine liver hydrolysates can be successfully incorporated into confectionery products to create functional gummies with potential health benefits, offering antioxidant protection and antimicrobial effects in a consumer-friendly form. Full article

Background/Objectives: A nanostructured membrane of polycaprolactone (a synthetic polymer) was synthesized using an electrospinning technique aiming to enhance its hydrophilicity and rate of degradation by surface modification via aminolysis. Since polycaprolactone nanofibrous films are naturally hydrophobic and with slow degradation, which restricts their use in biological systems, amino groups were added to the fiber surface using the aminolysis technique, greatly increasing the wettability of the membranes. Methods: Polycaprolactone nanofibrous membranes were synthesized via the electrospinning technique and surface modification by aminolysis. Trypsin, pepsin, and pancreatin were conjugated onto the aminolyzed PNF surface to further strengthen biocompatibility by enhancing the hydrophilicity, porosity, and biodegradation rate. SEM, FTIR, EDX, and liquid displacement method were performed to investigate proteolytic efficiency and morphological and physical characteristics such as hydrophilicity, porosity, and degradation rates. Results: Enzyme activity tests, which showed a zone of clearance, validated the successful enzyme conjugation and stability over a wide range of pH and temperatures. Scanning electron microscopy (SEM) confirms the smooth morphology of nanofibers with diameters ranging from 150 to 950 nm. Fourier transform infrared spectroscopy (FTIR) revealed the presence of O–H, C–O, C=O, C–N, C–H, and O–H functional groups. Energy-dispersive X-ray (EDX) elemental analysis indicates the presence of carbon, oxygen, and nitrogen atoms owing to the presence of peptide and amide bonds. The liquid displacement technique and contact angle proved that Pepsin-PNFs possess notably increased porosity (88.50% ± 0.31%) and hydrophilicity (57.6° ± 2.3 (L), 57.9° ± 2.5 (R)), respectively. Pancreatin-PNFs demonstrated enhanced enzyme activity and degradation rate on day 28 (34.61%). Conclusions: These enzyme-conjugated PNFs thus show improvements in physicochemical properties, making them ideal candidates for various biomedical applications. Future studies must aim for optimization of enzyme conjugation and in vitro and in vivo performance to investigate the versatility of these scaffolds. Full article