Physicochemical, antioxidant, antimicrobial, and in vitro cytotoxic activities of corn pollen protein hydrolysates obtained by different peptidases

The applications of protein hydrolysates as food preservatives and nutraceutical ingredients have attracted much attention because of their beneficial effects. The interest in these ingredients has shifted toward their biological activities with benefits to human health. Bioactive peptides are known as antioxidant agents that could promote health-promoting effects and prolong food shelf-life beyond their basic nutritional value. Thus, the aim of this study was to investigate antioxidant, antimicrobial, and in vitro cytotoxic properties of corn pollen protein (CPP) hydrolysates obtained by different enzymes. Proteolytic activity in terms of degree of hydrolysis (DH) and SDS-PAGE analysis was measured in pancreatin (H-Pan), pepsin (H-Pep), and trypsin (H-Tri) hydrolysates. Amino acid composition, antioxidant and antimicrobial activities, and cytotoxicity of hydrolysates were evaluated. DH and SDS-PAGE revealed higher proteolytic activity of pepsin compared to other enzymes. Amino acid analysis showed that the functional amino acids such as antioxidant types were most predominant in H-Pep compared to two other samples. Antioxidant activity of hydrolysates was found to be affected by the type of enzyme and the concentration of hydrolysates. There was a significant difference (p < 0.05) between antioxidant activity of different hydrolysates. The highest antioxidant activity in terms of Trolox equivalent antioxidant capacity (0.23–2.75 mM), DPPH (33.3%–64.8%), and hydroxyl (33.7%–63.2%) radical scavenging activities, chelation of iron (33.2%–62.5%) and copper (30.2%–50.5%) metals, and total antioxidant activity (0.65–0.85) was obtained for H-Pep followed by H-Pan and H-Tri samples. Antibacterial tests showed that pepsin-hydrolyzed protein was not significantly (P > 0.05) effective against E. coli at any concentrations, however, it showed significant (P < 0.05) concentration-dependent effect against S. aureus (with inhibition zones of 15–25 mm). Cytotoxicity results revealed that CPP, as a nonhydrolyzed protein, did not generally show antiproliferative activity, however, a significant (P < 0.05) ability of H-Pep hydrolysate in decreasing HT-29 colon cancer cell line viability was seen in a concentration-dependent manner (the lowest cell viability of 32% at 5 mg/mL). Overall, investigating the application of protein-based hydrolysates is one of the possible strategies that govern their applied intentions as preservatives and nutraceuticals in the food and pharmaceutical industries

Predictive modeling of oleuropein release from double nanoemulsions: An analytical study comparing intelligent models and Monte Carlo simulation

The objective of this study was to evaluate the release of oleuropein (OLP) from double nanoemulsions stabilized with polymeric complexes. Initially, W1/O nano-emulsions loaded with OLP were prepared and re-emulsified into an aqueous phase (W2), which included a complex of whey protein concentrate (WPC)/pectin, to form W1/O/W2 emulsions. The microstructure of the final double emulsions was analyzed by scanning electron microscopy (SEM), and particles with smooth, comparatively spherical, and somewhat asymmetrical surfaces with a size range of 100–200 nm were observed, which were compatible with dynamic light scattering (DLS) data. The release trend of OLP was determined by fitting it to several empirical models including zero order, first order, Higuchi, Hixson-Crowell, Korsemeyer-Peppas, Baker-Lonsdale, and utilizing intelligent modeling techniques such as Fuzzy Logic (FL) and Artificial Neural Networks (ANNs). Among the mathematical models, the zero order equation had the highest coefficient of determination (R2 = 0.988), while the first order equation had the lowest root-mean-square error (RMSE = 0.0176) and sum of squared errors (SSE = 0.0009) for the goodness of fit of the model, when considering the release trend of OLP. FL and ANNs proved effective in modeling controlled release of OLP-loaded nanocarriers, achieving high R2 values. Additionally, Monte Carlo (MC) simulation showed potential for evaluating the release process when compared to other methods

Enhancing the bio-polymeric stabilization of spray-dried Chlorella phenolic-rich extract: Analysis of its physicochemical, functional, structural, and biological properties

In this study, the spray-drying process of Chlorella extract was investigated. The influence of different carriers including maltodextrin (MD), acacia gum (AG), whey protein concentrate (WPC) on the physicochemical, functional, flowability and color properties was analyzed. Powder obtained using MD revealed the highest bulk and tapped density (0.613 and 0.750 g/mL, respectively), the highest solubility (96.87 %) and hygroscopicity (29.87 %). Dry powder attained by MD, showed the highest content of total phenols (0.413 mg GAE/mL) and lowest particle size (12.70 μm). All produced powders revealed fair to good cohesiveness and flowability. Besides, the spray-drying processes using (MD -WPC) showed higher maintenance of the antioxidant activity. SEM images revealed the presence of particles of different sizes and wrinkles. Evaluation of the chemical structure (FTIR) indicated the encapsulation and distribution of the phenolic compounds of the extract in the matrix of the carriers. Research findings can confirm the possibility to use encapsulated Chlorella extracts as an abundant source of natural antioxidants in the enrichment and formulation of various beneficial food products. Finally, the results showed the efficiency of the spray-drying process on the production of dry powder extract of Chlorella with appropriate physical properties, stability and antioxidant properties

Protection of navy-bean bioactive peptides within nanoliposomes: morphological, structural and biological changes

Abstract This study aimed to produce bioactive peptides from navy-bean protein with alcalase and pepsin enzymes (30–300 min) and to load them into a nanoliposome system to stabilize and improve their bioavailability. The degree of hydrolysis and biological activities (scavenging of DPPH, OH, and ABTS free radicals, reducing power, and chelating metal ions) of navy-bean protein were affected by the type of enzyme and hydrolysis time. The average particle size (83–116 nm), PDI (0.23–0.39), zeta potential (− 13 to − 20 mV), and encapsulation efficiency (80–91%) of nanoliposomes were influenced by the type and charge of peptides. The storage temperature and the type of loaded peptide greatly affected the physical stability of nanocarriers and maintaining EE during storage. The FTIR results suggested the effect of enzymatic hydrolysis on the secondary structures of protein and the effective placement of peptides inside polar-regions and the phospholipid monolayer membrane. SEM images showed relatively uniform-sized particles with irregular structures, which confirmed the results of DLS. The antioxidant activity of primary peptides affected the free radical scavenging of loaded nanoliposomes. Liposomes loaded with navy-bean peptides can be used as a health-giving formula in enriching all kinds of drinks, desserts, confectionery products, etc. Graphical Abstrac

Technological, nutritional, and biological properties of apricot kernel protein hydrolyzates affected by various commercial proteases

Abstract The effect of enzymatic hydrolysis of apricot kernel protein with different proteases (Alcalase, pancreatin, pepsin, and trypsin) on the amino acid content, degree of hydrolysis (DH), antioxidant, and antibacterial characteristics of the resulting hydrolyzates was investigated in this study. The composition of amino acids (hydrophobic: ~35%; antioxidant: ~13%), EAA/TAA ratio (~34%), and PER index (~1.85) indicates the ability of the hydrolyzate as a source of nutrients and antioxidants with high digestibility. Enzymatic hydrolysis with increasing DH (from 3.1 to a maximum of 37.9%) led to improved solubility (especially in the isoelectric range) and changes in water‐ and oil‐holding capacity. The highest free radical scavenging activity of DPPH (83.3%), ABTS (88.1%), TEAC (2.38 mM), OH (72.5%), NO (65.7%), antioxidant activity in emulsion and formation of TBARS (0.36 mg MDA/L), total antioxidant (1.61), reducing power (1.17), chelation of iron (87.7%), copper (34.8%) ions, and inhibition of the growth of Escherichia coli (16.3 mm) and Bacillus cereus (15.4 mm) were affected by the type of enzymes (especially Alcalase). This research showed that apricot kernel hydrolyzate could serve as a nutrient source, emulsifier, stabilizer, antioxidant, and natural antibacterial agent in functional food formulations

Modification of Whey Proteins by Sonication and Hydrolysis for the Emulsification and Spray Drying Encapsulation of Grape Seed Oil

In this study, whey protein concentrate (WPC) was sonicated or partially hydrolyzed by Alcalase, then examined as an emulsifier and carrier for the emulsification and spray drying of grape seed oil (GSO)-in-water emulsions. The modification treatments increased the free amino acid content and antioxidant activity (against DPPH and ABTS free radicals), as well as, the solubility, emulsifying, and foaming activities of WPC. The modified WPC-stabilized emulsions had smaller, more homogeneous droplets and a higher zeta potential as compared to intact WPC. The corresponding spray-dried powders also showed improved encapsulation efficiency, oxidative stability, reconstitution ability, flowability, solubility, and hygroscopicity. The morphology of particles obtained from the primary WPC (matrix type, irregular with surface pores) and modified WPC (reservoir type, wrinkled with surface indentations), as well as the oxidative stability of the GSO were influenced by the functional characteristics and antioxidant activity of the carriers. Changes in the secondary structures and amide regions of WPC, as well as the embedding of GSO in its matrix, were deduced from FTIR spectra after modifications. Partial enzymolysis had better results than ultrasonication; hence, the WPC hydrolysates are recommended as emulsifiers, carriers, and antioxidants for the delivery and protection of bioactive compounds

Stabilization of antioxidant thyme-leaves extract (Thymus vulgaris) within biopolymers and its application in functional bread formulation

The aim of this study was to stabilize the antioxidant compounds of thyme (Thymus vulgaris) extract by spray-drying process and MD, WPC and MD-WPC carriers at different temperatures (120 and 150°С). Then, the optimal encapsulated sample was used in different ratios (1, 2, 3 and 5% w/w) to fortify pan-breads. The highest production yield (∼68 %) was obtained in the extract encapsulated with MD-WPC at 150°С. The values of moisture (3.2–4.1 %) and water activity (0.28–0.33) indicated the microbial stability of the powders. The functional characteristics of the particles (solubility and wettability) were affected by the composition and nature of the carriers. Also, the use of WPC led to an increase in physical stability, encapsulation efficiency (∼84 %), TPC (4.4 mgGAE/g) and antioxidant activity (∼77 %). The results of SEM and staining of the samples respectively indicated the effect of the process parameters on the morphology and the placement of the phenolic compounds within carrier's matrix. The physicochemical, textural characteristics, color indices (crumb and crust) and antioxidant activity of fortified breads were affected by the ratio of added free and spray-dried extract. The evaluation of sensory indices (color, texture, chewiness, taste and overall acceptance) indicated the ability to fortify bread with 3 % spray-dried extract