Pomegranate seed as a novel source of plant protein: Optimization of protein extraction and evaluation of in vitro digestibility, functional, and thermal properties.

This research was carried out to optimize the extraction process of proteins from pomegranate seeds and characterize their in vitro digestibility as well as their thermal and functional properties. For this purpose, the study screened five parameters (liquid/solid ratio, pH, temperature, NaCl concentration, and time) that could potentially influence the extraction process. This screening was conducted using a two-level Placket-Burman design (PBD). The significant parameters (pH and NaCl concentration) were subsequently optimized using a three-level face-centered central composite design (FCCD) to determine the optimum extraction conditions. A maximum protein recovery of 83.8% was obtained at pH 11.0 and NaCl concentration of 0.0 M. Pomegranate seed protein isolate (PSPI) with a protein content of 92.4% (w/w) was obtained through the isoelectric precipitation of pomegranate seed protein extracted under the optimized conditions. An emulsifying activity index of 14.1 m2 g-1 was observed at the isoelectric pH, where the emulsion stability index was at 8.2%. PSPI also showed high water- and oil-holding capacities (3.7 and 4.3 g g-1, respectively). The essential amino acid levels in PSPI (except for valine and isoleucine) exceeded the recommended amounts set by WHO/FAO/UNU for adults, highlighting its high nutritional value. Based on thermal analysis data, denaturation of PSPI could occur at 89.5°C. The in vitro digestibility of PSPI was found to be 74.3%. PSPI shows a potential as a novel ingredient for substituting animal-based proteins in various food applications.

Fabrication and characterization of a succinyl mung bean protein and arabic gum complex coacervate for curcumin encapsulation.

The present study developed a novel complex coacervate based on succinyl mung bean protein (SMBP) and gum arabic (GA) to encapsulate curcumin. The optimum pH and succinylprotein/polysaccharide ratio for complex coacervation were 3.0 and 1:1, respectively, measured by turbidity evaluation. Fluorescence spectroscopy depicted that the curcumin was loaded in the hydrophobic core of SMBP/GA. The evaluated FTIR and XRD showed that the encapsulation of curcumin in the complex coacervate hydrophobic core was successful, followed by minor changes in SMBP conformation caused by the succinylation process. The zeta potential showed that the succinylation of MBP led to a decrease in the zeta potential of SMBP and confirmed that the SMBP/GA was produced successfully at pH 3.0. The EE and LA of c-SMBP/GA were 99.79 ± 0.03 % and 24.94 ± 0.05 µg·mg-1, respectively, which were significant. SMBP showed enhanced antioxidant activity compared with MBP, and c-MBP/GA showed significant antioxidant activity measured by ABTS and DPPH radical scavenging assays. SMBP is a biopolymer that can be used to encapsulate bioactive compounds like curcumin and shows enhanced antioxidant activity. The c-SMBP/GA is a promising tool for encapsulating curcumin in food matrices with enhanced dispersity characteristics and release behavior.

Curcumin-loaded complex coacervate made of mung bean protein isolate and succinylated chitosan as a novel medium for curcumin encapsulation.

A novel complex coacervate based on mung bean protein (MBP) and succinylated chitosan (SC) was developed in order to encapsulate curcumin to enhance its antioxidant and release properties. The optimum pH and MBP/SC ratio for fabrication of the complex coacervate were determined as 5.5 and 3:1, respectively. The MBP/SC complexes exhibited high affinity toward curcumin with encapsulation efficiency of 89.65%. The curcumin-loaded MBP with succinyl chitosan (c-MBP/SC) exhibited antioxidant properties investigated by DPPH and reducing power assays. c-MBP/SC also showed significant photo stability and acceptable controlled release behavior in simulated gastrointestinal conditions. Fluorescence results indicated that curcumin interacted with the hydrophobic areas available in c-MBP/SC. FTIR results showed the successful encapsulation of curcumin in the hydrophobic core of the complex, followed by minor changes in MBP conformation. Analysis of zeta potential revealed that MBP/SC particles were synthesized successfully at the pH value of 5.5 due to conformational changes of MBP. The conformational changes in protein structure were confirmed by Nile Red fluorescence anisotropy. As a result, c-MBP/SC could be considered as a promising carrier for curcumin encapsulation in food formulations with enhanced dispersity characteristic.

High and low oxalate content in spinach: an investigation of accumulation patterns.

BACKGROUND: Oxalic acid is a common antinutrient in the human diet, found in large quantities in spinach. However, spinach is highly regarded by vegetable producers because of its nutritional content and economic value. One of the primary purposes of spinach-breeding programs is to improve the nutritional value of spinach by adjusting oxalate accumulation. Knowledge of the biosynthetic patterns of oxalic acid, and its different forms, is important for a better understanding of this process. RESULTS: We found three biosynthetic patterns of accumulation and concentration of oxalates. Two of them are related to the maximum type and one is related to the minimum type. We also developed a general model of variations in these compounds in the genotypes that were studied. CONCLUSION: This study introduced a unique type of spinach with high oxalate accumulation, which could be particularly suitable for consumption. This had the highest ratio of insoluble oxalate to soluble oxalate. It also accumulated more ascorbic acid (AA) than other types. Our findings in this study also indicate a small role for AA as a precursor to oxalate production in spinach, possibly confirming the significant role of glyoxylate as the most critical precursor in this plant. © 2021 Society of Chemical Industry.

Epitope mapping and the effects of various factors on the immunoreactivity of main allergens in egg white.

Egg white has high protein content and numerous biological/functional properties. However, reported allergenicity for some of the proteins in egg white is an issue that needs to be paid exclusive attention. A consideration of the structure of IgE epitopes and their sequences, as well as a comprehensive understanding of the effects of various processes on epitopes and the impact of the gastrointestinal tract on them, can help target such issues. The current study focuses on the identified IgE epitopes in egg white proteins and evaluation of the effects of the gastrointestinal digestion, carbohydrate moiety, food matrix, microbial fermentation, recombinant allergen, heat treatment, Maillard reaction and combination of various processes and gastrointestinal digestion on egg white allergenicity. Although the gastrointestinal tract reduces the immunoreactivity of native egg white proteins, some of the IgE epitope-containing fragments remain intact during the digestion process. It has been found that the gastrointestinal tract can have both positive and negative impacts on the IgE binding activities of egg white proteins. Elimination of the carbohydrate moiety leads to a reduction in the immunoreactivity of ovalbumin. But, such effects from the carbohydrate parts in the IgE binding activity need to be explored further. In addition, the interaction between the egg white proteins and the food matrix leads to various effects from the gastrointestinal tract on the digestion of egg white proteins and their subsequent immunoreactivity. Further on this matter, studies have shown that both microbial fermentation and Maillard reaction can reduce the IgE binding activities of egg white proteins. Also, as an alternate approach, the thermal process can be used to treat the egg white proteins, which may result in the reduction or increase in their IgE binding activities depending on the conditions used in the process. Overall, based on the reported data, the allergenicity levels of egg white proteins can be mitigated or escalated depending on the conditions applied in the processing of the food products containing egg white. So far, no practical solutions have been reported to eliminate such allergenicity.

Improving the Biological Value of Olive and Soybean Oil Blends with Olive Leaf Extract Obtained by Ultrasound-Assisted Extraction towards the Preparation of a Sauce Product.

French sauce from different blends of soybean and olive oils was prepared and the oxidative stability of the optimum sauce sample, enriched with various amounts of olive leaf polyphenolic extract (OLE) (obtained via ultrasound-assisted extraction), was investigated over 90 days of storage. The microbiological and sensory properties of the samples containing the optimum amounts of OLE, as a substitution for synthetic preservatives, were studied. According to the results, the addition of olive oil at higher levels (75% and 100%) could affect the physicochemical properties of the sauce as compared to the control sample. It was also found that the addition of olive oil (up to 50%) would not significantly impact the sauce properties. Regarding the OLE enrichment in the samples, it was found that high levels of OLE could improve the oxidative stability of the samples. It was also found that OLE could be used as a preservative instead of commercial ones. Overall, this study suggests the potential use of olive oil and olive leaf extract in the preparation of French sauce to boost its nutritional value and its stability.

Effect of enzyme immobilization and in vitro digestion on the immune-reactivity and sequence of IgE epitopes in egg white proteins.

The effects of hydrolysis by free and immobilized forms of Neutrase (FN, IN, respectively) and Thermolysin (FT, IT, respectively) and in vitro digestion on the degree of hydrolysis (DH) of egg white proteins, molecular weight distribution of peptides, immune-reactivity and IgE epitopes of egg white proteins were investigated. With FT and IT in the intestinal digests, the proteolysis followed by in vitro digestion produced peptides smaller than 10 kDa. Hydrolysis with the immobilized enzymes had a greater effect than the free enzymes on increasing surface hydrophobicity. The lowest IgE-binding capacity was observed for the intestinal digest of IT-derived hydrolysates (3.3 ± 1.9%). Compared to in vitro digestion, proteolysis showed a significant effect on the immune-reactivity reduction of egg white proteins. Liquid chromatography-tandem mass spectrometry data showed that the most resistant epitopes to enzymatic hydrolysis and in vitro digestion were in ovomucoid, where epitope fragments 1-10, 1-14, 1-20, 4-20, 11-20, 61-74, 71-75 and 101-105 remained intact. Overall, the IgE-binding capacities of egg white proteins were not completely removed after the enzymatic hydrolysis and in vitro digestion due to the presence of intact proteins such as lysozyme and also due to the several immunoreactive peptides derived from egg white proteins.

Applying native proteases from melon to hydrolyze kilka fish proteins (Clupeonella cultriventris caspia) compared to commercial enzyme Alcalase.

The enzymatic hydrolysis (at 2.5, 5.0, 10.0, and 15.0% degree of hydrolysis, DH) of kilka fish (Clupeonella cultriventris caspia) was investigated using a crude melon extract (CME) as well as a commercial serine protease (Alcalase). Hydrolysates from both enzymatic treatments were analyzed for their antioxidant and functional properties. The hydrolysis resulted in increased antioxidant activities of kilka fish protein hydrolysates and the highest antioxidant activity was obtained for the CME hydrolysates with 5.0% DH level. Both treatments improved the protein solubility level to >65% within a pH range of 2.0-10.0. At a given DH level, CME hydrolysates showed higher oil and water holding capacities than Alcalase hydrolysates. Hydrolysates from CME exhibited better emulsifying properties compared to those prepared by Alcalase, particularly at low DH (2.5 and 5.0%). According to the results of this study, CME can be suggested as a new source of proteolytic enzymes for fish protein hydrolysis.

Identification of Potent ACE Inhibitory Peptides from Wild Almond Proteins.

In this study, the production, fractionation, purification and identification of ACE (angiotensin-I-converting enzyme) inhibitory peptides from wild almond (Amygdalus scoparia) proteins were investigated. Wild almond proteins were hydrolyzed using 5 different enzymes (pepsin, trypsin, chymotrypsin, alcalase and flavourzyme) and assayed for their ACE inhibitory activities. The degree of ACE inhibiting activity obtained after hydrolysis was found to be in the following order: alcalase > chymotrypsin > trypsin/pepsin > flavourzyme. The hydrolysates obtained from alcalase (IC50 = 0.8 mg/mL) were fractionated by sequential ultrafiltration at 10 and 3 kDa cutoff values and the most active fraction (<3 kDa) was further separated using reversed phase high-performance liquid chromatography (RP-HPLC). Peptide sequence identifications were carried out on highly potential fractions obtained from RP-HPLC by means of liquid chromatography coupled to electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS). Sequencing of ACE inhibitory peptides present in the fraction 26 of RP-HPLC resulted in the identification of 3 peptide sequences (VVNE, VVTR, and VVGVD) not reported previously in the literature. Sequence identification of fractions 40 and 42 from RP-HPLC, which showed the highest ACE inhibitory activities (84.1% and 86.9%, respectively), resulted in the identification of more than 40 potential ACE inhibitory sequences. The results indicate that wild almond protein is a rich source of potential antihypertensive peptides and can be suggested for applications in functional foods and drinks with respect to hindrance and mitigation of hypertension after in vivo assessment. PRACTICAL APPLICATION: This study has shown the potential of wild almond proteins as good sources for producing ACE-inhibitory active peptides. According to this finding, peptides with higher ACE inhibitory activities could be released during the gastrointestinal digestion and contribute to the health- promoting activities of this natural protein source.

Rheology and microstructure of kefiran and whey protein mixed gels.

The effect of kefiran on cold-set gelation of whey protein isolate (WPI) at 25 °C was studied using rheological measurements and environmental scanning electron microscopy (ESEM). The gelation of samples was induced by the addition of glucono-δ-lactone to the dispersions. WPI concentration was maintained at 8% (w/v) and the concentration of kefiran varied from 0 to 0.08% (w/v). According to rheological measurements, the addition of kefiran into WPI dispersions resulted in a significant increase in the gel strength, the yield stress, and the shear stress values at the flowing point. The gelling point and gelation pH of samples decreased significantly with an increase in kefiran concentration. ESEM micrographs showed that the presence of kefiran played an important role in the microstructure formation of gels. The microstructure of kefiran-WPI mixed gels was more compact and dense, compared to the WPI gel. Depletion interactions between kefiran and whey protein aggregates can be regarded as the chief factor which was responsible for these effects. The present work demonstrated that rheological and microstructural properties of acid-induced whey protein gels were improved by the addition of kefiran.

Extraction optimization and physicochemical properties of pectin from melon peel.

In this study, acidic extraction (citric acid) was used for the extraction of pectin from melon peel. A central composite design (CCD) was applied to study the effect of temperature (35-95°C), time (40-200min), pH (1-3) and solvent to sample ratio (10-50v/w) on the yield and degree of esterification (DE). The results showed that the yield and DE ranged from 2.87 to 28.98% and 1.33-29.33%, respectively. Also, the highest extraction yield (29.48±1.7%) was obtained under optimal conditions (pH of 1, temperature of 95°C and ratio of 10v/w after 200min). The protein, ash, carbohydrate and galacturonic acid content of pectin obtained under optimal conditions were about 1.5, 3.5, 10 and 48%, respectively. Rheological experiments determined that melon peel pectin dispersions behaved as a weak gel at concentrations of 1%w/v. The emulsifying activity was 35% and also, the emulsion stability was higher at 4°C than at 23°C.

Source identification of perylene in surface sediments and waterbird eggs in the Anzali Wetland, Iran.

Following the marked increase of perylene concentration in southern coast of Caspian Sea, waterbird eggs were used as biomonitoring agents. Surface sediments and eggs of five bird species were collected from colonies in Anzali Wetland in the above coast for perylene analysis. The perylene concentrations in sediment and egg samples ranged within 70.6-204.4 and 25.5-43.2 ng/g dw, respectively. Diagnostic perylene ratios showed that the perylene found in all samples was of biogenic origin, possibly developing from terrestrial materials. The combination pattern of perylene was found to be similar in all samples. Conclusively, perylene observed in the area was transmitted from sediments in breeding areas into the eggs, so the eggs are biomonitoring agents and the prevalence of oxic conditions in surface sediments limits formation of perylene, reflecting perylene formation in the catchment area. We found that perylene distribution in surface sediments follows irregular patterns, representing significant effects from local inputs.

Modelling of aflatoxin G1 reduction by kefir grain using response surface methodology.

Aflatoxin G1 (AFG1) is one of the main toxic contaminants in pistachio nuts and causes potential health hazards. Hence, AFG1 reduction is one of the main concerns in food safety. Kefir-grains contain symbiotic association of microorganisms well known for their aflatoxin decontamination effects. In this study, a central composite design (CCD) using response surface methodology (RSM) was applied to develop a model in order to predict AFG1 reduction in pistachio nuts by kefir-grain (already heated at 70 and 110°C). The independent variables were: toxin concentration (X1: 5, 10, 15, 20 and 25 ng/g), kefir-grain level (X2: 5, 10, 20, 10 and 25%), contact time (X3: 0, 2, 4, 6 and 8 h), and incubation temperature (X4: 20, 30, 40, 50 and 60°C). There was a significant reduction in AFG1 (p < 0.05) when pre-heat-treated kefir-grain used. The variables including X1, X3 and the interactions between X2-X4 as well as X3-X4 have significant effects on AFG1 reduction. The model provided a good prediction of AFG1 reduction under the assay conditions. Optimization was used to enhance the efficiency of kefir-grain on AFG1 reduction. The optimum conditions for the highest AFG1 reduction (96.8%) were predicted by the model as follows: toxin concentration = 20 ng/g, kefir-grain level = 10%, contact time = 6 h, and incubation temperature = 30°C which validated practically in six replications.

Stabilization of canthaxanthin produced by Dietzia natronolimnaea HS-1 with spray drying microencapsulation.

The strain bacterium Dietzia natronolimnaea has propounded as a source for biological production of canthaxanthin. Because of sensitivity of this pigment, examine on its stability is important. In this study, stability of encapsulated canthaxanthin from D. natronolimnaea HS-1 using soluble soybean polysaccharide (SSPS), gum acacia (GA), and maltodextrin (MD) as wall materials was investigated at 4, 25, and 45 °C in light and dark conditions during 4 months of storage. It was shown that the type of walls influenced the size of emulsion droplets; spray dried particles, microencapsulation efficiency (ME), and retention of canthaxanthin in microcapsules. SSPS and MD produced the smallest and the biggest emulsion droplets and spray dried particles, respectively. Microcapsules made with SSPS resulted in better ME and higher stability for canthaxanthin. Samples were degraded in all conditions, especially in light and 45 °C. Degradation of microencapsulated canthaxanthin with SSPS and GA proceeded more slowly than did with MD. Regardless of the type of wall materials, total canthaxanthin contents of the microencapsulated products decreased by an increase in time or temperature. Also, samples exposed to light indicated less stability at 4 and 25 °C when compared to the storage at dark conditions. According to the results of this study, SSPS can be considered as potential wall material for the encapsulation of carotenoids.

Investigation of different parameters on acrylamide production in the fried beef burger using Taguchi experimental design.

Acrylamide is a carcinogenic compound which is produced as a result of thermal processing of food materials such as French fries, cereals and meat products. In this study the effects of four different parameters on the level of produced acrylamide in two types of beef burgers during the frying was investigated. Each parameter was used in three levels (temperature at 170, 190, and 210 °C; frying time at 5, 6, and 7 min and meat level at 30, 60, and 85%, and also three types of oil, corn, canola and sunflower). Taguchi's L9 design was applied to carry out the experiments. While temperature and meat level indicated more effect on the production of acrylamide in the studied samples, type of oil did not show any significant effects at all. Frying time (within the range studied here) showed minor contribution on the acrylamide level produced during the frying.

General analytical schemes for the characterization of pectin-based edible gelled systems.

Pectin-based gelled systems have gained increasing attention for the design of newly developed food products. For this reason, the characterization of such formulas is a necessity in order to present scientific data and to introduce an appropriate finished product to the industry. Various analytical techniques are available for the evaluation of the systems formulated on the basis of pectin and the designed gel. In this paper, general analytical approaches for the characterization of pectin-based gelled systems were categorized into several subsections including physicochemical analysis, visual observation, textural/rheological measurement, microstructural image characterization, and psychorheological evaluation. Three-dimensional trials to assess correlations among microstructure, texture, and taste were also discussed. Practical examples of advanced objective techniques including experimental setups for small and large deformation rheological measurements and microstructural image analysis were presented in more details.

Antibacterial properties and chemical characterization of the essential oils from summer savory extracted by microwave-assisted hydrodistillation

Antibacterial properties and chemical characterization of the essential oils from summer savory (Satureja hortensis) extracted by microwave-assisted hydrodistillation (MAHD) were compared with those of the essential oils extracted using the traditional hydrodistillation (HD) method. While MAHD at 660 W required half as much time as HD needed, similar antibacterial efficacies were found from the essential oils obtained by the two extraction methods on two food pathogens (Staphylococcus aureus, a gram positive bacterium, and Escherchia coli, a gram negative bacterium). Also, as it was the case with the essential oils extracted by HD, that of MAHD indicated greater influence on S. aureus than on E. coli. The compositions of the extracted essential oils were also studied using GC-MS analysis. The same components with negligible differences in their quantities were found in the extracted essential oils using the two methods outlined above. Overall, to reduce the extraction time, MAHD can be applied at higher microwave levels without any compromise in the antibacterial properties of the essential oils extracted.

"A comparison between sugar consumption and ethanol production in wort by immobilized Saccharomyces Cerevisiae, Saccharomyces Ludwigii and Saccharomyces Rouxii on Brewer's Spent Grain"

The immobilization of Saccharomyces cerevisiae DSM 70424, Saccharomyces ludwigii DSM 3447 and Saccharomyces rouxii DSM 2531 on brewer's spent grain and then ethanol production and sugar consumption of these immobilized yeasts were investigated. The aim of this study was to investigate the abilities of these three immobilized yeasts for producing alcohol for brewing at two temperatures (7 and 12 °C) using two different sugar levels (one at original level supplied in the brewery and one with 2.5 percent (w/v), added glucose to the wort). Increasing both parameters resulted in higher alcohol production by all the yeasts studied. At 7 °C and with original wort density the ethanol content at the end of fermentation was 2.7 percent (v/v) for S. cerevisiae, 1.7 percent for S. ludwigii and 2.0 percent for S. rouxii. After the addition of 2.5 percent (w/v) glucose at the same temperature (7 °C), the alcohol production was increased to 4.1, 2.8 and 4.1 percent, respectively. Similar improvements were observed when the fermentation was carried out at 12 °C with/without the addition of glucose to the wort. However, temperature indicated greater influence on S. ludwigii than did on S. rouxii and S. cerevisiae. The immobilization as carried out in this study impacted both S. ludwigii and S. rouxii in a way that they could consume maltose under certain conditions.

On the formulation design and rheological evaluations of pectin-based functional gels.

In the current study, formulation design and development of a novel pectin-based functional gel were investigated. Amidated low methoxyl pectin (ALMP), high methoxyl pectin (HMP), sorbitol, inulin, rebaudioside-A, gardenia, phloridzin, quercetin, apple flavor, and calcium chloride (as its dihydrated salt) were selected as general ingredients. Response surface methodology was applied to design different formulations and to investigate on their experimental responses. The oscillatory tests were carried out in 2 stages with ALMP, HMP, sorbitol, and inulin as the variables of the 1st stage and ALMP and calcium chloride as the variables of the 2nd stage. Results of the 1st stage indicated relatively similar behaviors throughout the frequency range applied for all of the samples studied (true gels). However, magnitudes of the 5 rheological parameters of this study (storage modulus, loss modulus, loss tangent, complex modulus, and complex viscosity) were affected by different variables (ALMP, HMP, inulin, and sorbitol concentrations). Experimental results confirmed that sorbitol could be omitted from the formula while inulin and HMP could be used at their highest levels studied. At the 2nd stage, only 2 formulas indicated a strong gel behavior and other formulas showed typical behaviors of weaker gels or those of the concentrated solutions. At the conclusion of this study, a finished gelled product formula was suggested with the application of the best levels of the ingredients. The resultant gel was found to be set rapidly with no syneresis and showed a potential to be considered as a functional gelled dessert.

"A comparison between sugar consumption and ethanol production in wort by immobilized Saccharomyces Cerevisiae, Saccharomyces Ludwigii and Saccharomyces Rouxii on Brewer'S Spent Grain".

The immobilization of Saccharomyces cerevisiae DSM 70424, Saccharomyces ludwigii DSM 3447 and Saccharomyces rouxii DSM 2531 on brewer's spent grain and then ethanol production and sugar consumption of these immobilized yeasts were investigated. The aim of this study was to investigate the abilities of these three immobilized yeasts for producing alcohol for brewing at two temperatures (7 and 12 °C) using two different sugar levels (one at original level supplied in the brewery and one with 2.5% (w/v), added glucose to the wort). Increasing both parameters resulted in higher alcohol production by all the yeasts studied. At 7 °C and with original wort density the ethanol content at the end of fermentation was 2.7% (v/v) for S. cerevisiae, 1.7% for S. ludwigii and 2.0% for S. rouxii. After the addition of 2.5% (w/v) glucose at the same temperature (7 °C), the alcohol production was increased to 4.1, 2.8 and 4.1%, respectively. Similar improvements were observed when the fermentation was carried out at 12 °C with/without the addition of glucose to the wort. However, temperature indicated greater influence on S. ludwigii than did on S. rouxii and S. cerevisiae. The immobilization as carried out in this study impacted both S. ludwigii and S. rouxii in a way that they could consume maltose under certain conditions.