Changes in physicochemical properties, gel structure and in vitro digestion of marinated egg white gel during braising.

In this paper, changes in physicochemical properties, gel structure and in vitro digestion of marinated egg with spice or tea during braising were investigated. Results indicated that the moisture content and surface hydrophobicity of marinated egg white showed an overall decreased trend. The springiness of marinated egg white showed an increased trend, and the hardness in the late stage showed an increased trend. Microstructure showed that compact gel structures formed many holes during the braising. Intermolecular forces showed that ionic bonds and disulfide bonds played a dominant role in the marinated egg white. Secondary structure showed that the ß-turn showed a decreased trend, contrary to that of random coils and α-helices. Appropriate braising increased the digestibility of marinated egg white, but excessively long-time braising could reduce it. Both spice and tea braising could improve the gel strength of protein, and the tea braising was also slightly better than spice braising.

[Enzymatic hydrolysates of whey protein and chicken egg ppotein: production, physical-chemical and immunochemical characteristics].

Reducing the manifestations of food allergy by the inclusion of specialized foods in the nutrition of children and adults suffering from this disease is an important problem. The aim was to obtain and characterize in vitro food protein hydrolysates to evaluate their use in specialized foods with reduced potential allergenicity. MATERIAL AND METHODS: Whey protein concentrate (WPC) and chicken egg protein (CEP) and enzymes such as pancreatin and alkalase have been used. Proteolysis of proteins was carried out in an FA-10 fermenter for 3 hours at an enzyme : substrate ratio of 1:50 in dry matter, at optimal pH and temperature for pancreatin and alkalase. Enzymes were inactivated at +75 °C and fermentolizate was ultrafiltered. The solutions were concentrated by reverse osmosis and freeze-dried. The molecular weight distribution of the peptide fractions was evaluated by HPLC. Residual antigenicity was determined by the method of indirect enzyme-linked immunosorbent assay and expressed as the fold of antigenicity reduction relative to the original protein. RESULTS AND DISCUSSION: During WPC proteolysis with pancreatin the hydrolyzate was obtained with a fold reduction of antigenicity of 2.3×103 relative to the initial WPC. A decrease in antigenicity of 4.7×104 times was achieved with proteolysis of WPC by alkalase. The combination of WPC fermentolysis with pancreatin or alkalase followed by ultrafiltration reduced the content of high molecular weight peptides with a mass more than 8.7 kDa. The multiplicity of decrease in antigenicity with respect to the starting protein was 1.64×105 and 1.90×105, respectively. After repeated ultrafiltration the reduction in antigenicity of the obtained WPC alkalase or pancreatin hydrolysate was more than 1.0×106 and more than 5.0×105, respectively. The decrease in antigenicity of the CEP hydrolyzate obtained with proteolysis by alkalase and ultrafiltration compared to the initial CEP was 1.0×105 times, and 5.0×105 times when we used repeated ultrafiltration. CONCLUSION: A significant decrease in the content of high molecular weight peptides and a decrease in the antigenicity of peptide mixtures based on WPC and CEP to the values that permit their use in hypoallergenic products is achieved by combining proteolysis and double ultrafiltration through a UF10 membrane.

Influences of fermentation and ripening temperatures on the enzymatic activity and physicochemical and sensory properties of salted egg white sufu.

This study developed a novel form of sufu. Salted egg white sufu (SEWS) was produced by fermenting steamed salted egg white with douchi koji and ripening at 25°C, 35°C, or 45°C for 19 days. The results show that the protease activity of the koji reduced pronouncedly during fermentation, whereas the one in the SEWS initially increased but subsequently decreased. The total nitrogen and amino nitrogen contents and degree of protein hydrolysis increased during fermentation and decreased during preripening. SEWS, particularly processed at 35°C, contained more free amino acids, notably glutamic acid and leucine, than steamed salted egg white did. After processing, SEWS had darker colors, particularly when manufactured at higher temperatures, and hardness and springiness decreased. The 35°C and 45°C SEWS had higher sensory acceptability. In conclusion, ripening at 35°C for 19 days is recommended for producing this novel animal protein-based sufu.

Proteolytic pattern, protein breakdown and peptide production of ovomucin-depleted egg white processed with heat or pulsed electric fields at different pH.

Protein susceptibility to in vitro gastrointestinal digestion of ovomucin-depleted egg white (OdEW) adjusted to pH 4, 5, 7 and 9 and processed by heat (60 and 80 °C for 10 min) or pulsed electric fields (PEF) (1.4-1.8 kV/cm, 259-695 kJ/kg) was studied by assessing peptide production, proteolytic pattern, and the final peptide profile. Ovotransferrin was more susceptible to pepsin hydrolysis than lysozyme, with ovalbumin showing the highest proteolytic resistance. Ovalbumin was, however, hydrolyzed by pancreatin to produce a stable fragment. Heat treatment of OdEW solutions at 60 °C had little impact on protein susceptibility with the ovalbumin dimers formed having a comparable resistance to pepsinolysis as ovalbumin. Heating at 80 °C significantly enhanced protein susceptibility, as ovalbumin and protein aggregates formed were completely hydrolyzed within 30 min of pepsinolysis. Adjusting OdEW solution to pH 4 and treating with PEF at 695 kJ/kg enhanced protein susceptibility, similar to heat treatment at 80 °C, mainly owing to the enhanced enzymatic hydrolysis of ovalbumin. PEF processing can, therefore, increase protein digestion while minimizing protein aggregation, which will enhance protein functionality in egg whites.

Influence of high-intensity ultrasound on foaming and structural properties of egg white.

The effects of high-intensity ultrasound (20 kHz) at varying power pre-treatments (90, 120, 240, 360 and 480 W for 10 min) on foaming and structural properties of egg white were studied in this paper. The highest foaming ability (260.00%) was obtained after 360 W ultrasound treatment which was 4.9-fold to the control group. The lower viscosity and surface tension, and higher protein solubility indicated that the protein was easier to attach to the gas-liquid interface and generate molecular rearrangement. Moreover, the increased free sulfhydryl groups and surface hydrophobicity revealed that the protein adopted a loose structure after ultrasonic processing. SDS-PAGE data proved that subunit of poorly water-soluble ovomucin was declined. Scanning electron microscopy showed different microstructure with smaller aggregates and pore structure compared to non-treated egg white. These results presented more evidence of protein properties under ultrasonic environment, and broadened the application range of ultrasonic technique in food industry.

Prediction of heat-induced polymerization of different globular food proteins in mixtures with wheat gluten.

Egg, soy or whey protein co-exists with wheat gluten in different food products. Different protein types impact each other during heat treatment. A positive co-protein effect occurs when heat-induced polymerization of a mixture of proteins is more intense than that of the isolated proteins. The intrinsic protein characteristics of globular proteins which enhance polymerization in mixtures with gluten are unknown. In this report, a model was developed to predict potential co-protein effects in mixtures of gluten and globular proteins during heating at 100°C. A negative co-protein effect with addition of lysozyme, no co-protein effect with soy glycinin or egg yolk and positive co-protein effects with bovine serum albumin, (S-)ovalbumin, egg white, whole egg, defatted egg yolk, wheat albumins and wheat globulins were detected. The level of accessible free sulfhydryl groups and the surface hydrophobicity of unfolded globular proteins were the main characteristics in determining the co-protein effects in gluten mixtures.

Effects of different polysaccharides on the formation of egg yolk LDL complex nanogels for nutrient delivery.

Five polysaccharides, pectin, carboxymethyl cellulose (CMC), gum arabic, carrageenan and alginate, were studied to form complex nanogels with egg yolk low density lipoprotein (LDL). All nanogels were smaller than 85nm with high negative zeta potential, while LDL/carrageenan and LDL/alginate nanogels exhibited more heterogeneous size distribution. Fourier transform infrared spectrum suggested that hydrogen bonds, hydrophobic and electrostatic interactions were involved to form nanogels. Overall, significant expansion of nanogels was observed after encapsulation of curcumin, being studied as a model lipophilic nutrient. Fluorescence spectra evidenced that LDL provided non-polar microenvironment for curcumin and polysaccharides played an important role in the encapsulation process. All nanogels showed sustained release of curcumin under simulated gastrointestinal conditions. Furthermore, nanoscale, smooth and spherical ultrafine dry powders of nanogels were obtained by innovative nano spray drying technology. Our study indicated that LDL/polysaccharides may serve as potential oral delivery systems for lipophilic nutrients.

Herring roe protein has a high digestible indispensable amino acid score (DIAAS) using a dynamic in vitro gastrointestinal model.

It is hypothesized that the digestible indispensable amino acid score (DIAAS) can be determined based on dynamic in vitro gastrointestinal digestion experiments as replacement for invasive animal studies. We determined the in vitro DIAAS for immature herring eggs (roe) proteins in comparison with reference proteins. The true ileal digestibility of protein and indispensable amino acids (IAA) was measured under human conditions simulated in a gastrointestinal model (tiny-TIM). The in vitro true ileal digestibility of ovalbumin, cooked and raw chicken egg white, and casein was similar to that found in humans (r(2) = 0.96), providing a casual observation to support the validity of tiny-TIM. The digestibility of the immature herring egg proteins was 71% to 92%. The highest IAA digestibility was found for immature whole herring egg protein (55%-80%) in comparison to immature herring egg membrane and immature de-membraned herring protein (50%-70%). The DIAAS as recommended by FAO for children and adults, but measured in vitro, were 91% for immature whole herring egg protein (lysine first limiting), 71% for immature herring egg membrane protein (histidine first limiting), and 88% for immature herring egg de-membraned protein (sulfur AA first limiting). True ileal protein and amino acid digestibility can be determined in a dynamic gastrointestinal model, such as tiny-TIM, which can be used for estimating the DIAAS. Immature herring egg proteins, a previously underutilized resource, were determined to be an important and valuable source of IAA for human consumption.

Changes in physical and gelling properties of freeze-dried egg white as a result of temperature and relative humidity.

BACKGROUND: The quality of dried egg white with respect to functional properties can be affected by storage conditions. The effect of temperature and relative humidity (RH) on changes in colour and gelling properties in freeze-dried egg white (FDEW) during storage was investigated. RESULTS: The glass transition temperature (Tg ) of FDEW decreased with increasing % RH. The colour of FDEW stored at 60 °C was darker yellow than those at 40 and 25 °C, particularly at high % RH. RH had no effect on hardness and water-holding capacity (WHC) of gels made from FDEW stored at 25 °C for 1 week. However, hardness and WHC of gels from FDEW stored at higher temperatures; 40 °C, 48% RH and 60 °C, 11% RH dramatically increased. These results related to the differential scanning calorimeter thermograms which showed a broadening peak with lower enthalpy of protein denaturation. Moreover, the protein's SDS-PAGE pattern in the samples stored at high temperatures or RH levels showed protein aggregation. CONCLUSION: Storage of FDEW at high temperature and RH levels induced protein conformation changes. These have contributed to protein aggregation which affected the gelling properties of FDEW. © 2016 Society of Chemical Industry.

Food-grade protein-based nanoparticles and microparticles for bioactive delivery: fabrication, characterization, and utilization.

Proteins can be used to fabricate nanoparticles and microparticles suitable for use as delivery systems for bioactive compounds in pharmaceutical, food, cosmetic, and other products. Food proteins originate from various animal or vegetal sources and exhibit a wide diversity of molecular and physicochemical characteristics, e.g., molecular weight, conformation, flexibility, polarity, charge, isoelectric point, solubility, and interactions. As a result, protein particles can be assembled using numerous different preparation methods, from one or more types of protein or from a combination of a protein and another type of biopolymer (usually a polysaccharide). The final characteristics of the particles produced are determined by the proteins and/or polysaccharides used, as well as the fabrication techniques employed. This chapter provides an overview of the functional properties of food proteins that can be used to assemble nanoparticles and microparticles, the fabrication techniques available to create those particles, the factors that influence their stability, and their potential applications within the food industry.

Egg proteins as allergens and the effects of the food matrix and processing.

Hen eggs are an important and inexpensive source of high-quality proteins in the human diet. Egg, either as a whole or its constituents (egg yolk and white), is a key ingredient in many food products by virtue of its nutritional value and unique functional properties, such as emulsifying, foaming, and gelling. Nevertheless, egg is also known because of its allergenic potential and, in fact, it is the second most frequent source of allergic reactions, particularly in children. This review deals with the structural or functional properties of egg proteins that make them strong allergens. Their ability to sensitize and/or elicit allergic reactions is linked to their resistance to gastroduodenal digestion, which ultimately allows them to interact with the intestinal mucosa where absorption occurs. The factors that affect protein digestibility, whether increasing it, decreasing it, or inducing a different proteolysis pattern, and their influence on their capacity to induce or trigger an allergic reaction are discussed. Special attention is paid to the effect of the food matrix and the processing practices on the capacity of egg proteins to modulate the immune response.

Isolation and characterization of an ovoinhibitor, a multidomain Kazal-like inhibitor from Turkey (Meleagris gallopavo) seminal plasma.

Turkey seminal plasma contains three serine proteinase inhibitors. Two of them, with low molecular masses (6 kDa), were identified as single-domain Kazal-type inhibitors responsible for regulating acrosin activity. Our experimental objective was to isolate and characterize the inhibitor with the high molecular weight from turkey seminal plasma. The inhibitor was purified using hydrophobic interaction and affinity chromatography. Pure preparations of the inhibitor were used for identification by mass spectrometry, for determination of physicochemical properties (molecular weight, pI, and content and composition of the carbohydrate component), for kinetic studies, and for antibacterial tests. Gene expression and immunohistochemical detection of the inhibitor were analyzed in the testis, epididymis, and ductus deferens. The inhibitor with a high molecular weight from turkey seminal plasma was identified as an ovoinhibitor, which was found in avian semen for the first time. The turkey seminal plasma ovoinhibitor was a six-tandem homologous Kazal-type domain serine proteinase inhibitor that targeted multiple proteases, including subtilisin, trypsin, and elastase, but not acrosin. Our results suggested that hepatocyte growth factor activator was a potential target proteinase for the ovoinhibitor in turkey seminal plasma. The presence of the ovoinhibitor within the turkey reproductive tract suggested that its role was to maintain a microenvironment for sperm in the epididymis and ductus deferens. The turkey seminal plasma ovoinhibitor appeared to play a significant role in an antibacterial semen defense against Bacillus subtilis and Staphylococcus aureus.

Inert and oxidative subcritical water hydrolysis of insoluble egg yolk granular protein, functional properties, and comparison to enzymatic hydrolysis.

The use of enzymes to recover soluble peptides with functional properties from insoluble proteins could prove to be very expensive, implying high reaction times and low yields. In this study, the insoluble granular protein, previously delipidated, was hydrolyzed using enzymes (trypsin) as a comparison to the proposed alternative method: subcritical water hydrolysis (SWH) using both nitrogen and oxygen streams. The result of the hydrolysis was characterized in terms of the yield and peptide size distribution as well as different functional properties. The SWH of the delipidated granules resulted in a higher recovery yield than that obtained by enzymatic hydrolysis in half of the time. The foaming capacity of the peptides obtained by SWH was higher than that obtained by trypsin hydrolysis, although the foam stability was lower. Slight differences were detected between these peptides in terms of their emulsifying properties.

Increased muscular triglyceride content and hyperglycemia in Goto-Kakizaki rat are decreased by egg white hydrolysate.

We investigated the fat metabolic characteristics in non-obese and diabetic Goto-Kakizaki (GK) rat and the effects of dietary egg white hydrolysate (EWH) on glucose and fat metabolism. Wistar (W) and GK (G) rats were placed into dietary casein (WC and GC) or EWH (WE and GE) group, and fed their respective diet for six weeks. Triglyceride (TG) content and stearoyl-CoA desaturase (SCD) indices in the soleus muscle were higher in the GC group than WC group in parallel with worsening serum glucose metabolic parameters. The glucose metabolic parameters were significantly improved in the GE group. The TG accumulation and SCD indices in the soleus muscle were also significantly lower in the GE group than in the GC group. In conclusion, dietary EWH not only improved glucose metabolism but also reduced both TG accumulation and SCD indices in the soleus muscle of GK rat.

Egg yolk: structures, functionalities and processes.

Hen egg yolk is an ideal example of natural supramolecular assemblies of lipids and proteins with different organization levels. These assemblies are mainly due to interactions between proteins and phospholipids, and these interactions are essential in understanding and controlling the production of food made with yolk, and particularly emulsions. Furthermore, these assemblies can be modulated by external constraints among which thermo-mechanical and high-pressure treatments. This review focuses on multi-scale structures present in egg yolk, and their modulation by processes, in relation with their emulsifying properties. Egg yolk is mainly composed of two fractions-plasma and granules-which are natural nano- and micro-assemblies. These two fractions possess different composition, structures and functionalities and exhibit specific behaviour under treatments such as high pressure and temperature. Plasma contains a large quantity of lipids structured as lipoproteins (low-density lipoproteins), whereas granules are mainly composed of proteins aggregated in micrometric assemblies. If plasma is responsible for the important emulsifying properties of yolk, granules bring interesting emulsifying properties when assemblies are in the form of micelles in presence of salts. High-pressure or thermal treatments, applied before or after emulsion fabrication, alter their functionalities and could be used to commercially exploit these fractions.

Isolation and characterization of domain I of ovoinhibitor.

Domain I of ovoinhibitor was isolated by subjecting the protein to specific chemical cleavage by cyanogen bromide followed by repeated gel filtration. The first domain of ovoinhibitor was found to be homogeneous by the criteria of gel chromatography, SDS-PAGE and PAGE. Mr values by gel filtration (10900) and SDS-PAGE (8300) were slightly higher than that computed from amino-acid sequence. This discrepancy has been attributed to the glycoprotein nature of domain I as it was found to contain 10% neutral carbohydrate and 2% sialic acid. Fluorescence spectral properties showed the presence of tryptophan in domain I. The amino-acid composition of domain I isolated in this study was in very good agreement with that computed from amino-acid sequence. Gel filtration behaviour of the first domain was consistent with a Stokes radius of 1.6 nm and a frictional ratio of 1.2 suggesting asymmetry and/or excessive hydration. Domain I was found to be a potent inhibitor of bovine trypsin but was virtually devoid of activity against chymotrypsin, elastase and proteinase K. The equilibrium association constant for domain I-trypsin complex was computed to be 6.6x10(8)M-1.

Vitellogenin and lipovitellin: zinc proteins of Xenopus laevis oocytes.

Xenopus laevis vitellogenin is a plasma protein that contains a total of 5 mol of metal/440 kDa dimer, 2 mol of zinc, and 3 mol of calcium (Montorzi et al. (1994) Biochem. Biophys. Res. Commun. 200, 1407-1413]. There are no other group IIB or transition metals in the molecule. The zinc atoms are removed instantaneously by 1,10-phenanthroline (OP) (pK 4.8). Once internalized by receptor-mediated endocytosis, vitellogenin is cleaved into multiple polypeptides, i.e., the two lipovitellin subunits (1 and 2) plus phosvitin; these are then stored as microcrystals within yolk platelets. We here show by metal analysis of the individual proteins generated by vitellogenin processing that zinc and calcium occur in different domains of the vitellogenin polypeptide chain. All of the vitellogenin zinc is present in lipovitellin, in amounts equal to 1 mol of zinc/141 kDa. Calcium, in contrast, is detected exclusively in phosvitin which, in addition, contains 3 mol of magnesium/35 kDa, apparently acquired following vitellogenin entry into the oocyte. The zinc in lipovitellin is removed by OP in a concentration-dependent manner with a pK of 4.8, identical to that obtained for vitellogenin, and by exposure to acidic conditions (below pH 5). Following removal of zinc, the two lipovitellin subunits remain associated, suggesting that zinc is not involved in their interaction. On exposure to 1% SDS, lipovitellin does dissociate into 106 and 33 kDa subunits. The presence of stoichiometric quantities of zinc in both vitellogenin and lipovitellin calls for the study of the hitherto unrecognized biochemistry and functions of these proteins in zinc metabolism and development of the frog oocyte and embryo.

The lipoprotein crystals in yolk platelets of a shark, Squalus ocanthias (Selachii).

Lipovitellin-phosvitin crystals in yolk platelets from the egg of a shark, Squalus ocanthias (Selachii), were studied by the selected area electron diffraction method. Ultra-thin sections of yolk platelets possess an orthorhombic lattice with lattice parameters: a = 8.0 nm, b = 15.9 nm, and c = 18.1 nm. Comparative and quality analysis of the reflection intensities of electron diffraction patterns between shark and other vertebrate yolk platelets suggest evolutionary variability of the lipovitellin structure. The lattice and its parameters, however, are highly conserved. The results of the present study fill the last gap in the comparative yolk platelet research which has been carried out over many years for exponents of all vertebrate taxonomic groups, from cyclostomes to reptiles.

Sequence of lamprey vitellogenin. Implications for the lipovitellin crystal structure.

The amino acid sequence of lamprey vitellogenin has been predicted from the nucleotide sequence of cloned cDNA. The sites of proteolytic cleavage that produce the lipovitellin complex from the vitellogenin have been located by comparing the N-terminal sequences of two lamprey lipovitellin polypeptides with the predicted sequence. These results also confirm that the vitellogenin sequence derived here corresponds to the lipovitellin complex for which the crystal structure has been solved previously. Predictions of secondary structure indicate that the region most likely to correspond to the large alpha-helical domain of the crystallographic model consists of vitellogenin residues 300 to 600. Similar to the lipovitellins of Xenopus laevis, lamprey lipovitellin appears to lack approximately 200 C-terminal residues that are present in vitellogenin. However, the lamprey lipovitellin differs from those of Xenopus and chicken in two respects. First, most of the serine-rich domain that is present as the phosvitin polypeptide in the lipovitellins of the higher vertebrates appears to be lost in the maturation of lamprey vitellogenin to lipovitellin. Second, the domains that constitute the large lipovitellin-1 polypeptide in Xenopus and chicken are present in two polypeptides in lamprey, owing to an additional proteolytic processing event.