Search Results (44)

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

Rice bean is a legume that is considered to be an important source of protein; it is still poorly understood but can produce peptides with various biological activities. These peptides can be released by enzymes during gastrointestinal passage and have a physiological effect on cancer cells. Since both the type of legume and the enzyme cocktail used are crucial for the production of peptides against certain cancer cell types, the anticancer effects of Vigna umbellata isolate and hydrolysate against MDA and SiHa cells were investigated in this study. The isolate was obtained via isoelectric precipitation, and the hydrolysate was obtained via pepsin P7012 and pancreatin P1750. The anticancer activity was determined by cell viability via the MTT assay, and a Pearson correlation coefficient was calculated to determine the relationships between the anticancer activity and total phenolic compounds, and the antioxidant activity of the isolated rice bean protein and its hydrolysate. The rice bean hydrolysate had a higher concentration of phenolic compounds than the isolate does, resulting in high antioxidant activity. The inhibition of proliferation in SiHa cells and MDA cells was twofold greater when they were exposed to the hydrolysate than when they were exposed to the isolate, with the IC₅₀ values at a concentration of 1000 μg/mL after 48 h of exposure for both MDA and SiHa cells. Rice bean protein hydrolysate is a good alternative for inhibiting the proliferation of SiHa cells and MDA cells, although further studies are needed. Full article

The current study focuses on evaluating the enzyme inhibition (acetylcholinesterase, α-amylase, α-glucosidase, pancreatin lipase), antioxidant, and antimicrobial activities of the “Modesto” apricot, “Stanley” plum, and their hybrid the “Stendesto” plum–apricot. The “Stendesto” is the only successful plum–apricot hybrid in Bulgaria. A spectrophotometric approach was used to evaluate the antioxidant activity following four complementary assays (DPPH, ABTS, FRAP, and CUPRAC). The “Stendesto” plum–apricot revealed its enhanced antioxidant potential compared to its parental lines. Apart from the free phenolics extraction, two other techniques (alkaline and acid hydrolysis) were applied to reveal the biological potential of the studied fruit. Not only free but also bound phenolic extracts were able to inhibit α-glucosidase and acetylcholinesterase, while none of the extracts inhibited lipase or α-amylase. None of the apricot extracts had antimicrobial activity, while the other fruit had limited antimicrobial activity. The proposed results undoubtedly reveal that hybrid fruits possess enhanced biological activity compared to their parents. This is a first comprehensive evaluation of hybrid fruits with reference to parental lines. This makes them an interesting research topic that should be better explored. Full article

Gastrointestinal disorders dysregulate the biochemical environment of the gastrointestinal tract by altering pH conditions during the gastric phase of digestion or by reducing the secretion of pancreatin during the intestinal part of the process. Ingested functional food could therefore lose some of its health-promoting potential apart from its nutritional value. In this work, we aimed to manufacture bread marked by decreased gluten content, using a commercial or laboratory sourdough, that could be appropriate for patients afflicted with wheat allergy, hypertension and pancreatic malfunctions. A reference sample (no sourdough) was prepared alongside wheat and wheat–rye bread samples—produced with either commercial or laboratory sourdough (L. plantarum BS, L. brevis 1269, L. sanfranciscensis 20663). We measured the QQQPP allergen content (ELISA) in bread extracts digested in vitro and determined how these extracted components affect the level of active angiotensin and alpha amylase (spectrophotometry). We then elucidated how these properties changed when physiological digestion conditions (pH and pancreatin activity) were disturbed to mimic gastric hyperacidity, hypochlorhydria or exocrine pancreatic insufficiency. The key finding was that every tested type of bread produced with laboratory sourdough exhibited pronounced angiotensin-converting enzyme inhibition. The effect was preserved even in dysregulated digestive conditions. The use of laboratory sourdough prevented an increase in allergenicity when pancreatin was restricted as opposed to the commercial sourdough, which surpassed the reference sample reading at 50% pancreatin. No statistically consistent link was reported when the inhibition of alpha amylase was assayed. In conclusion, functional bread manufactured with sourdough composed of L. plantarum BS, L. brevis 1269, and L. sanfranciscensis 20663 was shown to be potentially capable of contributing to the treatment against hypertension as evidenced by in vitro research. It was also moderately safer with regard to its allergenicity. Full article

Bioactive peptides derived from native proteins modulate physiological processes in the metabolic pathways. Given that multiple protocols in the literature mimic the digestion of dietary components, gathering studies that use such models directed at protein digestion processes is critical. This systematic review aimed to gather evidence that adopted adequate experimental models to simulate human protein digestion. The databases searched were PubMed, Web of Science, ScienceDirect, Embase, Virtual Health Library, and Scopus. A total of 1985 articles were found, resulting in 20 eligible in vitro studies. The Office of Health Assessment and Translation was used to evaluate methodological quality. Seven studies used plant-based protein sources, twelve used animal protein sources, and one used both. The duration of the oral phase varied, although 60% of the studies employed a protein digestion period of 120 min. Amylase, pepsin, and pancreatin enzymes were utilized in 40% of the studies, with pH levels of 7, 3, and 7, respectively, during the oral, gastric, and intestinal phases. The INFOGEST harmonized static model was adopted by 65% of the studies; INFOGEST is the most effective model for simulating gastrointestinal protein processes in humans and can be used to answer several research questions because it describes experimental conditions close to the human physiological situation. Full article

This study aims to investigate the probiotic properties of various isolated strains of Lactiplantibacillus plantarum. Specifically, the focus is on examining the expression of the glutamic acid decarboxylase (GAD) gene and its role in the production of gamma-aminobutyric acid (GABA), a promising postbiotic metabolite. The investigation includes comprehensive analyses of morphology, genetics, resilience against bile, NaCl, and simulated pancreatin juice (SPJ), carbohydrate fermentation patterns, antibacterial activity, susceptibility to antibiotics, and the presence of β-D-galactosidase and GAD enzymes. Six L. plantarum strains exhibited remarkable resilience against bile, NaCl, and SPJ, as well as susceptibility to antibiotics and antagonistic behavior against pathogens. These strains also showed the presence of β-D-galactosidase. Additionally, five L. plantarum strains were found to harbor the gad gene. Further biochemical analysis of four specific L. plantarum strains revealed promising profiles consisting of antibiotics, vitamins, hormones, and a diverse array of metabolites with potential immunotherapeutic properties. This study highlights the substantial potential of Lactiplantibacillus plantarum in generating beneficial postbiotic metabolites. The identified strains offer exciting avenues for further exploration, with potential applications in functional foods and pharmaceuticals. This research opens up possibilities for harnessing the probiotic and postbiotic potential of L. plantarum to develop novel products with health-promoting properties. Full article

Tableting of biomolecules is a challenging formulation phase due to their sensitivity to various process parameters, such as compression pressure, process dynamics, or the temperature generated. In the present study, pancreatin was employed as a model enzyme mixture, which was formulated in tablet form utilizing the synergistic effects of brittle and plastic excipients (dibasic calcium phosphate and microcrystalline cellulose, respectively). The effect of varying compaction pressure and lubricant concentration on the generated temperature and enzymatic activity was evaluated. The tablets were analyzed for pancreatin content and the activity of two enzymes (protease and amylase) using pharmacopoeial tests. This study indicated that the formulations proposed here allow tableting over a wide range of compaction pressures without adversely affecting pancreatin content and its enzymatic activity. Full article

The regulation and characterization of nanomaterials in foods are of great interest due to the potential risks associated with their exposure and the increasing number of applications where they are used within the food industry. One factor limiting the scientifically rigorous regulation of nanoparticles in foods is the lack of standardized procedures for the extraction of nanoparticles (NPs) from complex matrices without alteration of their physico-chemical properties. To this end, we tested and optimized two sample preparation approaches (enzymatic- and alkaline-based hydrolyses) in order to extract 40 nm of Ag NP, following their equilibration with a fatty ground beef matrix. NPs were characterized using single particle inductively coupled plasma mass spectrometry (SP-ICP-MS). Fast sample processing times (<20 min) were achieved using ultrasonication to accelerate the matrix degradation. NP losses during the sample preparation were minimized by optimizing the choice of enzymes/chemicals, the use of surfactants, and the product concentration and sonication. The alkaline approach using TMAH (tetramethylammonium hydroxide) was found to have the highest recoveries (over 90%); however, processed samples were found to be less stable than the samples processed using an enzymatic digestion based upon pork pancreatin and lipase (≈60 % recovery). Low method detection limits (MDLs) of 4.8 × 10⁶ particles g⁻¹ with a size detection limit (SDL) of 10.9 nm were achieved for the enzymatic extraction whereas an MDL of 5.7 × 10⁷ particles g⁻¹ and an SDL of 10.5 nm were obtained for the alkaline hydrolysis. Full article

This study was undertaken to investigate the enzymatic hydrolysis of lentil starch concentrates from conventional cooked seeds (CCLSC) by the action of different types of enzymes, including pancreatin (PC-EHSC), heat-stable α-amylase (HS-EHSC), β-amylase (βA-EHSC), amyloglucosidase (AMG-EHSC), and multi-enzymes (βA-HS-AMG-EHSC); their multi-scale structural characteristics of the enzymatic hydrolysis products of lentil starch concentrates were compared. The morphological features distinguished among different samples. The Fourier-transform infrared spectroscopy and solid-state ¹³C CP/MAS NMR spectral features indicated the possible formation of a binary and ternary complex among amylose, protein and lipids. The X-ray diffraction results revealed that the V-type characteristic diffraction peaks were more obvious for samples including PC-EHSC and βA-EHSC, which was in line with their lowest polydispersity index (DP_n). PC-EHSC and βA-EHSC also showed an increased peak intensity of the scattering maximum on the small-angle X-ray scattering spectra, whereas CCLSC exhibited an overall lower peak intensity within the studied q range of scattering. The highest XRD crystallinity and the lowest DP_n value obtained for PC-EHSC indicated that the starch polymers modified by pancreatin could produce glucan chains with a comparatively homogenous M_w distribution that are readily recrystallized by hydrogen bonding through chain aggregation. Comparatively, the lowest relative crystallinity for HS-EHSC obtained from XRD suggested that thermostable α-amylolysis was unfavorable for the formation of starch structure with a higher degree of molecular order. This study could provide useful information for the needed research to obtain a deeper understanding of the impact of different amylolysis actions on the structural organization of starch hydrolysates and to provide a theoretical foundation for the development of fermentable enzymatically hydrolyzed starch with well-tailored physiological properties. Full article

In vitro digestion methods that can accurately predict the estimated GI (eGI) values of complex carbohydrate foods, including biscuits, are worth exploring. In the current study, standard commercial biscuits with varied clinical GI values between 9~30 were digested using both the INFOGEST and single-enzyme digestion protocols. The digestion kinetic parameters were acquired through mathematical fitting by mathematical kinetics models. The results showed that compared with the INFOGEST protocol, the $AU{R}_{180}$ deduced from digesting using either porcine pancreatin or α-amylase showed the best potential in predicting the eGI values. Accordingly, mathematical equations were established based on the relations between the $AU{R}_{180}$ and the GI values. When digesting using porcine pancreatin, $GI=$ 1.834 + 0.009 $\times AUC{R}_{180}$ ($R^2=$ 0.952), and when digesting using only α-amylase, $GI=$ 6.101 + 0.009 $\times AUC{R}_{180}$ $(R^2=$0.902). The $AU{R}_{180}$ represents the area under the curve of the reducing-sugar content normalized to the total carbohydrates versus the digestion time in 180 min. The in vitro method presented enabled the rapid and accurate prediction of the eGI values of biscuits, and the validity of the formula was verified by another batch of biscuits with a known GI, and the error rate of most samples was less than 30%. Full article

Arthrospira platensis contains high protein content and quality, which makes it a promising feed source for livestock animals. However, this microalga presents a recalcitrant peptidoglycan cell wall, and its main proteins form protein-pigment complexes attached to the algal thylakoid membrane. The purpose of the present study was to evaluate the effect of mechanical/physical pretreatments (bead milling, extrusion, freeze-drying, heating, microwave and sonication) combined with commercial enzymes (trypsin or pancreatin) on the degradation of A. platensis proteins. Protein degradation was assessed through the quantification of total protein and protein fractions (18–26 kDa, 40–48 kDa and others) on SDS-PAGE gels and the evaluation of the coefficient of protein degradation (CPD). The results showed that no significant differences were found among treatments for CPD values, except for an increase related to 18–26 kDa protein (phycocyanin subunits) with extrusion combined with pancreatin. In addition, extrusion and microwave caused a decrease of total protein in algal supernatant probably due to a denaturation/aggregation and reduction of solubility. Overall, extrusion is a promising pretreatment for A. platensis cell wall disruption and protein bioaccessibility. Further studies could elucidate how the effect of extrusion on protein solubility affects the activity of peptidases on protein degradation. Full article

The germination of lamtoro gung has been shown to increase the angiotensin-I-converting enzyme inhibitory (ACE-I) activity in previous studies. The 48 h germinated flour had the highest ACE-I activity. Administration of the gastrointestinal digestion (GID) simulation with commercial enzymes was expected to increase the ACE-I activity. However, the GID simulation to increase ACE-I in the germinated lamtoro gung flour has not been found. Therefore, this study aimed to evaluate the GID simulation of ACE-I peptides in sprouted lamtoro gung flour. This study also identified and characterised the peptide with the ACE-I activity. The GID simulation was performed using commercial pepsin (pH 2) and pancreatin (pH 7.5). Both simulations occurred at 37 °C for 240 min. The degree of hydrolysis, peptide concentration, and ACE-I activity was analysed. Samples with the highest ACE-I activity were then fractionated and identified, to determine the peptide responsible for the ACE-I activity. The 180 min GID simulation in the test sample showed the highest ACE-I activity (89.70%). This result was supported by an increased degree of hydrolysis (DH) and peptide concentrations throughout the GID simulation. The <1 kDa peptide fraction had the highest inhibitory activity and had the most elevated peptide portion (54.69%). Peptide sequences containing crucial amino acids were found in the <1 kDa peptide fraction. PRPPKPP, PPPPPGARAP, and PFPPSNPPP had proline in the C and N terminal residues. The peptides obtained also had other biological activities, such as a DPP IV inhibitor, an alpha-glucosidase inhibitor, and antioxidative activity. Based on the toxicity prediction, those peptides are non-toxic and safe to consume. Full article

Alpha-lactalbumin is a whey protein that is a cheese-making industrial residue of high biological value. The antihypertensive capacity of three peptides obtained from the simulated gastrointestinal digestion of alpha-lactalbumin hydrolysates was studied. The alpha-lactalbumin hydrolysis was performed using the Alcalase enzyme and was subsequently subjected to a simulated digestion process using pepsin and pancreatin enzymes to mimic digestion conditions. The peptides were identified from a RP-HPLC fractionation of the digest and subsequent identification by mass spectrometry analysis. Three peptides from the alpha-lactalbumin sequence were obtained: IWCKDDQNPH (P1), KFLDDDLTDDIM (P2), and DKFLDDDLTDDIM (P3). The in vitro antihypertensive activity of the peptides was determined by studying the inhibition of the angiotensin-converting enzyme, with P1 being the only peptide with antihypertensive activity detected by this methodology (IC₅₀ = 3.91 ± 0.2 mg/mL). In order to correlate the structural (molecular dynamics simulations) and physicochemical properties with potential mechanisms of antihypertensive capacity, in silico methods were performed. The peptides P1, P2, and P3 had a negative global charge and were hydrophilic. After molecular modeling, the peptide structures were submitted to a refinement based on an energy minimization and further molecular dynamics simulation to assess their global size and conformational space. After a 50-nanosecond simulation, the global structures, solvated and immersed in an ionic water solution similar to that of blood, were studied in their solvent-accessible surfaces. A secondary structure (alpha-helix) was observed in the P1 peptide, but in general, all peptides showed an extended folding. The surfaces were charge code colored and in a visual inspection it could be conjectured that all of them exposed the charge, mainly a negative charge, to the solvent surface, in agreement with the GRAVY index, which was also evaluated. In conclusion, the structure and amino acid composition of peptide 1 assessed by in silico studies agrees with the antihypertensive activity obtained by the in vitro study. Full article

Jackfruit leaf protein concentrate (LPC) was hydrolyzed by pepsin (H–Pep) and pancreatin (H–Pan) at different hydrolysis times (30–240 min). The effect of the enzyme type and hydrolysis time of the LPC on the amino acid composition, structure, and thermal properties and its relationship with the formation of O/W emulsions were investigated. The highest release of amino acids (AA) occurred at 240 min for both enzymes. H–Pan showed the greatest content of essential and hydrophobic amino acids. Low β-sheet fractions and high β-turn contents had a greater influence on the emulsifier properties. In H–Pep, the β-sheet fraction increased, while in H–Pan it decreased as a function of hydrolysis time. The temperatures of glass transition and decomposition were highest in H–Pep due to the high content of β-sheets. The stabilized emulsions with H–Pan (180 min of hydrolysis) showed homogeneous distributions and smaller particle sizes. The changes in the secondary structure and AA composition of the protein hydrolysates by the effect of enzyme type and hydrolysis time influenced the emulsifying properties. However, further research is needed to explore the use of H–Pan as an alternative to conventional emulsifiers or ingredients in functional foods. Full article

The health benefits of probiotics are beyond doubt. The positive effects of lactobacilli and bifidobacteria on the function of many body systems have been repeatedly proven by various studies. To completely realize the potential of probiotic microorganisms, the strains should be tested by the greatest combination of characteristics that contribute to the wellness of the host. In this work, for the first time, a combined assessment of the probiotic properties and vitamin B-producing potential of various species and strains of bifidobacteria and lactobacilli was carried out. The presence of an additional advantage, such as vitamin-producing ability, can prevent vitamin deficiency both at the level of the consumption of fermented foods, when the enrichment will occur naturally on the spot, and during colonization by these intestinal strains, when synthesis will occur in vivo. To select potential probiotics, the stress tolerance ability of 16 lactic acid bacteria and bifidobacteria strains to low pH values, bile, and proteolytic enzymes, as well as their ability to autoaggregate, were studied under conditions of modeling the gastrointestinal tract in vitro. The ability of the strains to extracellularly accumulate water-soluble B vitamins was evaluated by capillary electrophoresis. Among the tested strains of bifidobacteria, B. adolescentis VKPM AC-1662 is of interest; it was characterized by the greatest stress tolerance ability and the ability to autoaggregate, in addition to the extracellular synthesis of riboflavin and pyridoxine. Among lactic acid bacteria, L. sakei VKPM B-8936 demonstrated the greatest tolerance to low pH, L. plantarum VKPM B–11007 to duodenal conditions, L. acidophilus VKPM B-2213 to pepsin, and L. salivarius VKPM B–2214 to pancreatin. The highest percentage of autoaggregation was observed in L. salivarius VKPM B-2214, which also accumulated the largest amount of pantothenic acid, but it was sensitive to stress conditions. The obtained results could be used to create new products enriched with probiotics and B vitamins. Full article