Optimization of extraction parameters of protein isolate from milk thistle seed: Physicochemical and functional characteristics.

In the current study, optimization of milk thistle protein extraction parameters was carried out in terms of purity and yield. In addition, the characterization of proteins isolated from milk thistle seeds was conducted. The optimal conditions for achieving the highest purity of protein (MTP) from milk thistle seeds were identified as extraction pH 9.47, temperature 30°C, and extraction time 180 min. Conversely, optimal values for overall protein yield (MTY) were determined at extraction pH 12, temperature 50°C, and extraction time 167 min. The proteins obtained under these two sets of conditions (MTP and MTY) demonstrated comparable oil absorption capacity (OAC), foaming, and emulsifying capabilities, as well as stability, aligning with findings from previous studies on seed protein. Both proteins had the highest protein solubilities at pH 11. Both proteins' zeta potentials were closest to zero at pH 4, demonstrating their closeness to the isoelectric point. MTP and MTY had poorer antioxidant capabilities than the other protein isolates/concentrates. MTP and MTY contain high ß sheet concentrations that might enhance thermal stability and lower the digestibility of proteins. In conclusion, the protein extraction process demonstrated a high potential for achieving both substantial yield and remarkable purity with some decent technological and functional properties, thus holding promise for various applications in diverse fields.

Utilisation of an active branching sucrase from Lactobacillus kunkeei AP-37 to produce techno-functional poly-oligosaccharides.

Glucansucrase AP-37 was extracted from the culture supernatant of Lactobacillus kunkeei AP-37 and characteristics of the glucan produced by the active glucansucrase in terms of structural and functional roles were determined in this study. A molecular weight around 300 kDa was observed for glucansucrase AP-37 and its acceptor reactions with maltose, melibiose and mannose were also conducted to unveil the prebiotic potential of the poly-oligosaccharides formed via these reactions. The core structure of glucan AP-37 was determined by 1H and 13C NMR and GC/MS analysis which revealed that glucan AP-37 was a highly branched dextran composing of high levels of (1 â†’ 3)-linked α-d-glucose units with low levels of (1 â†’ 2)-linked α-d-glucose units. The structural features of the glucan formed, demonstrated that glucansucrase AP-37 was an α-(1 â†’ 3) branching sucrase. Dextran AP-37 was further characterised by FTIR analysis and XRD analysis demonstrated its amorphous nature. A fibrous compact morphology was observed for dextran AP-37 with SEM analysis whereas TGA and DSC analysis revealed its high stability as no degradation was observed up to 312 °C. Finally, the prebiotic potential of the dextran AP-37 and the gluco-oligosaccharides produced with the acceptor reaction of α-(1 â†’ 3) branching sucrase AP-37 were determined and promising results were found for the gluco-oligosaccharides to act as prebiotics.

Egg Yolk-Free Vegan Mayonnaise Preparation from Pickering Emulsion Stabilized by Gum Nanoparticles with or without Loading Olive Pomace Extracts.

The yolk-free mayonnaise was formed by Pickering emulsions stabilized by free and encapsulated olive pomace extracts (OPEs) in rocket seed [rocket seed gum nanoparticle (RSGNP)] and chia seed gum nanoparticles at different nanoparticle concentrations. The yolk-free mayonnaise and the control mayonnaise samples were compared in terms of appearance, microstructural, droplet size, emulsion stability, rheological, oxidative stability, and sensory properties. The droplet size decreased by increasing the nanoparticle concentration in yolk-free mayonnaise samples. The yolk-free mayonnaise samples prepared with OPE-loaded gum nanoparticle showed shear-thinning, solid-like and recoverable characteristics, which increased as the increase in the nanoparticle concentration. The emulsion stability and capacity increased by increasing the nanoparticle concentration in the yolk-free mayonnaise samples. OPE-loaded gum nanoparticle-stabilized yolk-free mayonnaise samples exhibited higher IP (induction period) values than the control samples. OPE-RSGNP 1% mayonnaise was observed to be the closest sample to the control sample with its sensory properties, general acceptability, and similar microstructural and rheological properties. The results of this study indicated that Pickering emulsions stabilized by gum nanoparticles could be used as healthy alternatives to the egg yolk in conventional mayonnaise.

The effects of different protease treatments on the techno-functional, structural, and bioactive properties of bovine casein.

In this study, bovine sodium caseinate (NaCas) was hydrolyzed with four proteases, alcalase, savinase, subtilisin A, and flavourzyme. In addition to the structural changes occurred through the enzymatic hydrolysis, the solubility, oil binding capacity, zeta potential, emulsification properties, and in vitro antioxidant capacity, anti-carcinogenic and antidiabetic properties of hydrolysates were determined. FTIR combined with hierarchical cluster analysis (HCA) made in Amide I region enable to classification of the samples based on the changes of the secondary structure depending on the enzyme type and degree of fragmentation. Technological properties of NaCas were enhanced through the enzymatic hydrolysis, and those were more prominent in serine-type enzymes, regardless of the enzyme type, all hydrolysates showed high antioxidant capacities. All hydrolysates, specifically those produced by savinase and alcalase, reduced the viability of the carcinogenic Caco-2 cells in a dose-dependent manner and showed a very low level of cytotoxicity against healthy HEK-293 cells. The hydrolysis treatment made a significant contribution to the antidiabetic activity of NaCas. Particularly alcalase and savinase hydrolysates suppressed the activity of α- amylase and α- glucosidase. Therefore, the generated milk protein hydrolysates could be used in functional food developments for specific dietary purposes.

Extraction of Natural Gum from Cold-Pressed Chia Seed, Flaxseed, and Rocket Seed Oil By-Product and Application in Low Fat Vegan Mayonnaise.

This study involves the modeling of rheological behavior of the gum solution obtained from cold-pressed chia seed (CSG), flaxseed (FSG), and rocket seed (RSG) oil by-products and the application of these gums in a low-fat vegan mayonnaise formulation as fat replacers and emulsifier. CSG, FSG, and RSG solutions showed shear-thinning flow behavior at all concentrations. The K values ranged between 0.209 and 49.028 Pa·sn for CSG, FSG, and RSG solutions and significantly increased with increased gum concentration. The percentage recovery for the G' was significantly affected by gum type and concentrations. CSG, FSG, and RSG showed a solid-like structure, and the storage modulus (G') was higher than the loss modulus (G″) in all frequency ranges. The rheological characterization indicated that CSG, FSG, and RSG could be evaluated as thickeners and gelling agents in the food industry. In addition, the rheological properties, zeta potential, and particle size and oxidative stability (at 90 °C) of low-fat vegan mayonnaise samples prepared with CSG, FSG, and RSG were compared to samples prepared with guar gum (GG), Arabic gum (AG), and xanthan gum (XG). As a result, CSG, FSG, and RSG could be utilized for low-fat vegan mayonnaise as fat and egg replacers, stabilizers, and oxidative agents. The results of this study indicated that this study could offer a new perspective in adding value to flaxseed, chia seed, and rocket seed cold-press oil by-product.

Production and characterization of yeast extracts produced by Saccharomyces cerevisiae, Saccharomyces boulardii and Kluyveromyces marxianus.

In recent years, prejudice in society against monosodium glutamate (MSG) has directed food manufacturers to alternative sources. Yeast extracts are considered as "natural" due to the production process and stand out due to their nutritional properties as well as giving a flavor similar to MSG. In this study, chemical, functional and flavor properties of yeast extract powders produced from Saccharomyces cerevisiae TGM10, Saccharomyces boulardii S11 and Kluyveromyces marxianus TGM66 were evaluated. Results revealed that the most protein-rich sample was S. cerevisiae TGM10 extract (69.17%), followed by S. boulardii S11 (66.16%) and K. marxianus TGM66 (62.42%) extracts, respectively and S. cerevisiae TGM10 extract was also the richest yeast extract for essential amino acids. Additionally, flavor-enhancing amino acids such as glutamic acid, aspartic acid, alanine and glycine were dominant in S. cerevisiae TGM10 extract (47.41 g/100 g protein). Sensorial evaluation of yeast extracts demonstrated that salty taste, umami taste and meaty flavor scores of yeast extracts were lower than MSG whereas for fruity flavor, yeast extracts had the highest scores. These findings revealed the potential of three yeast strains to produce yeast extracts in order to increase the nutritional value and flavor of foods.

Electrosprayed chitosan-coated alginate-pectin beads as potential system for colon-targeted delivery of ellagic acid.

BACKGROUND: Ellagic acid (EA), a potent dietary antioxidant, has limited bioavailability owing to its rapid absorption in the stomach and small intestine, and EA is transformed to more bioavailable compounds - urolithins - in the colon. An encapsulation system that sustains the release of EA in the gastrointestinal system and delivers more EA into the colon could improve the oral bioavailability of EA. Electrosprayed EA-loaded alginate-pectin beads were produced and coated with low- (LC) and high-molecular-weight chitosan (HC). The EA release from uncoated and coated beads under simulated gastrointestinal conditions was evaluated. The samples were characterized by particle size, gel strength, scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) analysis. RESULTS: The encapsulation efficiency (EE%) of EA ranged from 49.53% to 69.85% for uncoated beads, which was elevated up to 86.50% by coating, and LC coating provided higher EE%. Pectin addition to alginate and chitosan coating reduced the gel strength and changed the size depending on the molecular weight of chitosan. SEM images of pectin-added beads showed fewer cracks but more wrinkles, and chitosan coating presented more aggregated surfaces. The ionic interaction of alginate-pectin-chitosan and the entrapment of EA were confirmed by FTIR. In the gastric medium, EA release was very low from uncoated beads (15.2-19.8%), and totally restricted by chitosan coating. In the intestinal stage, EA release from LC-coated alginate-pectin beads was only 18%, and it was between 55% and 65% for uncoated or HC-coated counterparts. CONCLUSION: The LC-coated alginate-pectin beads could be further explored as a potential system for colon-targeted delivery of EA. © 2021 Society of Chemical Industry.

Encapsulation of Olive Pomace Extract in Rocket Seed Gum and Chia Seed Gum Nanoparticles: Characterization, Antioxidant Activity and Oxidative Stability.

The aim of this study was to determine the potential use of rocket seed and chia seed gum as wall materials, to encapsulate and to prevent degradation of olive pomace extract (OPE) in polymeric nanoparticles, and to characterize olive pomace extract-loaded rocket seed gum nanoparticles (RSGNPs) and chia seed gum nanoparticles (CSGNPs). The phenolic profile of olive pomace extract and physicochemical properties of olive pomace, rocket seed gum (RSG), and chia seed gum (CSG) were determined. The characterization of the nanoparticles was performed using particle size and zeta potential measurement, differential scanning calorimeter (DSC), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), encapsulation efficiency (EE%), in vitro release, and antioxidant activity analyses. Nanoparticles were used to form oil in water Pickering emulsions and were evaluated by oxitest. The RSGNPs and CSGNPs showed spherical shape in irregular form, had an average size 318 ± 3.11 nm and 490 ± 8.67 nm, and zeta potential values of -22.6 ± 1.23 and -29.9 ± 2.57, 25 respectively. The encapsulation efficiency of the RSGNPs and CSGNPs were found to be 67.01 ± 4.29% and 82.86 ± 4.13%, respectively. The OPE-RSGNP and OPE-CSGNP presented peaks at the 1248 cm-1 and 1350 cm-1 which represented that C-O groups and deformation of OH, respectively, shifted compared to the OPE (1252.53 cm-1 and 1394.69 cm-1). The shift in wave numbers showed interactions of a phenolic compound of OPE within the RSG and CSG, respectively. In vitro release study showed that the encapsulation of OPE in RSGNPs and CSGNPs led to a delay of the OPE released in physiological pH. The total phenolic content and antioxidant capacity of RSGNPs and CSGNPs increased when the OPE-loaded RSGNPs and CSGNPs were formed. The encapsulation of OPE in RSGNPs and CSGNPs and the IP values of the oil in water Pickering emulsions containing OPE-RSGNPs and OPE-CSGNPs were higher than OPE, proving that OPE-loaded RSGNPs and CSGNPs significantly increased oxidative stability of Pickering emulsions. These results suggest that the RSG and CSG could have the potential to be utilized as wall materials for nanoencapsulation and prevent degradation of cold-pressed olive pomace phenolic extract.

A new combination strategy to enhance apoptosis in cancer cells by using nanoparticles as biocompatible drug delivery carriers.

Some experimental and clinical studies have been conducted for the usage of chemotherapeutic drugs encapsulated into nanoparticles (NPs). However, no study has been conducted so far on the co-encapsulation of doxorubicin (Dox) and epoxomicin (Epo) into NPs as biocompatible drug delivery carriers. Therefore, we investigated if co-encapsulation of doxorubicin (Dox) and/or epoxomicin (Epo) into NPs enhance their anticancer efficiency and prevent drug resistance and toxicity to normal cells. We synthesized Dox and/or Epo loaded poly (lactic-co-glycolic acid) (PLGA) NPs using a multiple emulsion solvent evaporation technique and characterized them in terms of their particle size and stability, surface, molecular, thermal, encapsulation efficiency and in vitro release properties. We studied the effects of drug encapsulated NPs on cellular accumulation, intracellular drug levels, oxidative stress status, cellular viability, drug resistance, 20S proteasome activity, cytosolic Nuclear Factor Kappa B (NF-κB-p65), and apoptosis in breast cancer and normal cells. Our results proved that the nanoparticles we synthesized were thermally stable possessing higher encapsulation efficiency and particle stability. Thermal, morphological and molecular analyses demonstrated the presence of Dox and/or Epo within NPs, indicating that they were successfully loaded. Cell line assays proved that Dox and Epo loaded NPs were less cytotoxic to single-layer normal HUVECs than free Dox and Epo, suggesting that the NPs would be biocompatible drug delivery carriers. The apoptotic index of free Dox and Epo increased 50% through their encapsulation into NPs, proving combination strategy to enhance apoptosis in breast cancer cells. Our results demonstrated that the co-encapsulation of Dox and Epo within NPs would be a promising treatment strategy to overcome multidrug resistance and toxicity to normal tissues that can be studied in further in vivo and clinical studies in breast cancer.

Home-made cheese preservation using sodium alginate based on edible film incorporating essential oils.

The objective of the present study is developing a new technique for the preservation of natural cheese by the use of an edible biofilm based on sodium alginate in order to evaluate the effect of the essential oils (O. basilicum L, R. officinalis L. A. herba alba Asso. M. pulegium L.) incorporated in the film on the oxidation stability, microbial spoilage, physicochemical characteristics and sensory criteria. The cheese samples coated with sodium alginate incorporated by the oils showed moderate stability in terms of oxidative stabilities of proteins and lipids during storage. In addition, poor microbial growth (total aerobic mesophilic flora, yeasts and fecal coliforms) was observed in cheese samples coated with biofilm, also, the growth of Staphylococci Salmonella and Molds for all types of cheese were completely inhibited. Additionally, it was observed that the biofilm coating reduced the weight loss and hardness of the cheese comparing with the uncoated sample. The results of sensory analysis revealed that uncoated cheese, coated with sodium alginate and sodium alginate composed of oil of O. basilicum were the most preferred by panelists, in comparison with others. Therefore, it was concluded that this technique of coating cheese with edible film activated with essential oils is preferred and favorable by virtue of the effect of oils preserving the cheese without seriously affecting their organoleptic properties.

Extraction of a novel water-soluble gum from nettle (Urtica dioica) seeds: Optimization and characterization.

Nettle seed gum (NSG) which is a novel potential source of hydrocolloid was characterized in terms of yield, physicochemical, rheological, functional and thermal properties. According to Response Surface Methodology, the maximum extraction yield was determined as 6.17% on dry basis. In order to clarify the structural information of the gum, the FT-IR analysis and monosaccharide composition determination were performed. In the results, the NSG had 8.48% moisture, 10.81% total ash, 2.89% total protein, 1.15% total oil and 76.67% total carbohydrate. The FT-IR analysis revealed the existence of carboxyl groups, which gives the gum the capability of ion binding. Furthermore, Mannose/Galactose ratio was calculated as 1.07, indicating the water solubility of gum. DSC analysis showed that the NSG was highly stable in thermal process. Intrinsic viscosity, average viscosity and molecular weight values were determined as 8.56 ± 0.4 dL/g and 1.89 ± 0.1 × 106 g/mol, respectively. The emulsification capacity, emulsion stability and foam formation capacity of the NSG were 48%, 40% and 4%, respectively. The NSG exhibited shear thinning behavior while dynamic rheological analysis indicated the NSG had viscoelastic properties and G' (the storage modulus) was higher than G″ (the loss modulus). In conclusion, the present study supplied novel knowledge on the NSG, which will be inestimable for clarifying its matchless functional properties.

Development and characterization of sodium alginate based active edible films incorporated with essential oils of some medicinal plants.

The aim of this study was to develop sodium alginate based active films incorporated with essential oils (EO) of R. officinalis L, A. herba alba Asso, O. basilicum L and M. pulegium L. the films were characterized in terms of bioactive properties including thickness, moisture content, water vapor and oxygen permeability, release test, mechanical, molecular, biodegradability and thermal properties. The results showed that the active films had a strong antibacterial effect against the six pathogenic bacteria with the inhibition zone between18.5 and 38.67 mm. furthermore, the antioxidant capacity of the films ranged from 4.57% to 23.09%. According to results of release test, physical, molecular, mechanical, biodegradability, thermal and barrier properties, the EOs were uniformly dispersed in the polymer matrix and improved slightly thermal (Tm) and barrier properties, while decreased tensile strength and it was obtained that this film is biodegradable in the soil. In conclusion, it could be said that sodium alginate based edible films incorporated with EO have a potential to be used food packaging applications.

Production and characterization of a new biodegradable fenugreek seed gum based active nanocomposite film reinforced with nanoclays.

In the present work, fenugreek seed gum (FSG)/clay nanocomposite films were prepared with nanoclays (Na+ montmorillonite [MMT], halloysite [HNT] and Nanomer® I.44 P [NM]) at different amounts (0, 2.5, 5.0 and 7.5g clay/100g FSG) by solution casting method and characterized. Increasing amount of nanoclay significantly (P<0.05) improved oxygen barrier and thermal properties of the biodegradable films. Agar diffusion tests revealed that FSG based nanocomposite films exhibited strong antimicrobial properties against foodborne pathogens namely Listeria monocytogenes, Escherichia coli O157:H7, Staphylococcus aureus and Bacillus cereus independently of clay type and concentration. In the case of mechanical properties, nanoclay incorporation up to 5% provided higher (P<0.05) tensile strength (TS) properties while elongation at break (EB) values of the films significantly (P<0.05) decreased in the presence of clay in the film matrix. SEM micrographs showed that especially lower levels (up to 5%) of nanoclay reinforcements provided a homogeneous and smooth film structure. In conclusion, FSG based nanocomposite films reinforced with nanoclays up to 5% showed a precious potential to be used in antimicrobial food packaging applications.

Anti-inflammatory and antiapoptotic effect of interleukine-18 binding protein on the spinal cord ischemia-reperfusion injury.

We investigated the anti-inflammatory and antiapoptotic effects of interleukin-18 binding protein (IL-18BP) on spinal cord ischemia/reperfusion (I/R) injury in rats. Twenty-one adult male rats were divided into three groups: sham, I/R, and I/R+IL-18BP. Proinflammatory cytokines were determined in rat blood samples by using ELISA, while apoptosis was immunohistochemically evaluated in spinal cord tissue using caspase 3. Both IL-18 and TNF-α were significantly decreased in the IL-18BP group compared to that in the sham group. The highest caspase 3 levels were observed in the I/R group, while the lowest levels were found in the sham group. The mean Tarlov score of the I/R group was significantly lower than that of the sham group. However, the mean Tarlov score of the IL-18BP group was significantly higher than that of the I/R group. The results of the current study demonstrate that IL-18BP plays both anti-inflammatory and antiapoptotic roles in spinal cord I/R injury.

In vitro/in vivo comparison of cefuroxime release from poly(ε-caprolactone)-calcium sulfate implants for osteomyelitis treatment.

This study aimed to investigate the release of cefuroxime axetil (CF) and calcium from poly(ε-caprolactone) (PCL)-calcium sulfate (CaS) implants (PCL:CaS 2:1-10% CF; PCL:CaS 2:1-20% CF; PCL:CaS 1:1-10% CF) for treating infectious bone diseases. Bioactivity, crystallinity and strength, and release profiles under standard and pressurized release conditions were studied. PCL:CaS 2:1-20% CF had slower release than 10% loading. These groups had no significant change in CF and Ca release in response to pressure. The PCL:CaS 1:1 group had the slowest release despite having higher CaS, probably due to more compaction of discs. In contrast, pressure caused significant differentiation of CF and Ca(2+) release. The presence of CaS enhanced mechanical properties and bioactivity of discs. SEM and XPS results showed calcium-phosphate containing accumulations on surfaces upon SBF incubation. CF-loaded implants were applied in a rabbit osteomyelitis model. In vivo CF release was enhanced with increased CaS proportions, suggesting that in vivo release conditions are closer to pressurized in vitro conditions. In the control group, there was still some inflammation in the bone and no complete coverage with bone was achieved in the defect site. Discs provided a suitable surface for regeneration of bone. However, bone formation in the PCL:CaS 1:1 disc implanted group was more complete and regular than in the 2:1 group.