Pilot-scale formation of whey protein aggregates determine the stability of heat-treated whey protein solutions—Effect of pH and protein concentration

Denaturation and consequent aggregation in whey protein solutions is critical to product functionality during processing. Solutions of whey protein isolate (WPI) prepared at 1, 4, 8, and 12% (wt/wt) and pH 6.2, 6.7, or 7.2 were subjected to heat treatment (85°C × 30 s) using a pilot-scale heat exchanger. The effects of heat treatment on whey protein denaturation and aggregation were determined by chromatography, particle size, turbidity, and rheological analyses. The influence of pH and WPI concentration during heat treatment on the thermal stability of the resulting dispersions was also investigated. Whey protein isolate solutions heated at pH 6.2 were more extensively denatured, had a greater proportion of insoluble aggregates, higher particle size and turbidity, and were significantly less heat-stable than equivalent samples prepared at pH 6.7 and 7.2. The effects of WPI concentration on denaturation/aggregation behavior were more apparent at higher pH where the stabilizing effects of charge repulsion became increasingly influential. Solutions containing 12% (wt/wt) WPI had significantly higher apparent viscosities, at each pH, compared with lower protein concentrations, with solutions prepared at pH 6.2 forming a gel. Smaller average particle size and a higher proportion of soluble aggregates in WPI solutions, pre-heated at pH 6.7 and 7.2, resulted in improved thermal stability on subsequent heating. Higher pH during secondary heating also increased thermal stability. This study offers insight into the interactive effects of pH and whey protein concentration during pilot-scale processing and demonstrates how protein functionality can be controlled through manipulation of these factors.     

Determination of protein in foods: comparison of net protein and crude protein (N × 6.25) values

The purpose of the present study was: (1) to determine net (true) protein (NP) values as sums of amino acid residues and to compare them with crude protein (PA) values; and (2) to calculate the nitrogen-to-NP conversion factors by means of linear regression analyses between the NP values and nitrogen content. The differences between NP and PA values varied between food groups. The NP values in milk and milk products were, on average, 5.5% smaller than the corresponding PA values. The corresponding figures for other foods were: meat and meat products 15.4%, fish and fish products 20.6%, cereals and bakery products 12.2%, vegetables, fruits and berries 13.9% and miscellaneous processed foods 16.1%. In the present study the nitrogen-to-NP conversion factor for milk and milk products was 5.94, meat and meat products 5.17, fish and fish products 4.94, cereals and bakery products 5.40, vegetables, fruits and berries 5.36 and miscellaneous processed foods 5.51; the general conversion factor for all foods was 5.33. The above respective factors were 5.0, 17.3,21.0, 13.6, 14.2, 11.8 and 14.7% smaller than the official factor of 6.25. We conclude that the traditional PA values (N × 6.25) deviate significantly from the NP values. We recommend that protein definitions and methods of determination should be re-evaluated in the appropriate international organizations and updated in line with current knowledge and analytical capabilities for both scientific and other purposes.     

Characterization of protein–polyphenol interactions

Dietary polyphenols have received attention for their biologically significant functions as antioxidants, anticarcinogens or antimutagens, which have led to their recognition as potential nutraceuticals. Polyphenols also characteristically possess a significant binding affinity for proteins, which can lead to the formation of soluble and insoluble protein–polyphenol complexes. Questions remain concerning whether and to what extent the protein–polyphenol interaction influences functionality. For example, is the formation of protein–polyphenol complexes an obstacle to the nutritional bioavailability of either species? This article discusses the development of suitable methodologies to investigate the physicochemical basis of protein–polyphenol interactions and the influence of structure–activity relationships on binding affinities.     

Rheology of galactomannan–whey protein mixed systems

The rheological behaviour of whey protein/galactomannan mixtures in aqueous solutions was studied under gelling conditions of the protein component, at neutral pH and at a pH close to the protein isoelectric point. The presence of the neutral polysaccharide had significant effects on the formation and viscoelastic behaviour of the cured gels. This effect was dependent on the structural organisation of the protein network. At pH 7 the galactomannan had a general positive effect on WPI gel formation. It is suggested that under these conditions, the protein network forms a continuous phase that accommodate the polysaccharide chains, acting as a filler of the protein network. The minimum protein concentration for gelation to occur, the gelation temperature and time all decrease in the presence of the galactomannan. Under pH conditions near the whey protein isoelectric point, different effects were observed as a result of the galactomannan addition. At low WPI concentration, the galactomannan had a detrimental influence on the protein network formation, but a negligible effect or even a positive influence on the gelation process at higher concentrations.     

Evaluation of pea protein–polysaccharide matrices for encapsulation of acid-sensitive bacteria

Protein–polysaccharide capsules containing Bifidobacterium adolescentis were produced and tested in a series of in vitro survival experiments to evaluate capsule protection of the bacterium to simulated stomach conditions, as well as their ability to release the encapsulated bacteria under conditions similar to those found in the lower gut. A protein fraction isolated from peas (pea protein isolate: PPI; 2.0%; w/v) was mixed with each of three different polysaccharides (0.5% (w/v) of either sodium alginate, iota-carrageenan and gellan gum) to produce capsules ranging in size from 2 to 3 mm diameter. All capsule formulations provided significant protection for cells exposed to synthetic stomach juice at 37 °C relative to non-encapsulated bacteria. In addition, PPI-alginate and PPI-iota-carrageenan capsules were found to dissolve in simulated intestinal fluid at 37 °C, releasing 70–79% of their bacteria “payload” within 3 h, with higher cell numbers being released from the freeze-dried capsules. PPI-gellan gum capsules did not dissolve to the same extent and the number of released cells was ~ 26–30% lower. Following a temporal rat feeding study with the test bacterium encapsulated in PPI-alginate, B. adolescentis-specific PCR and qPCR analyses confirmed the presence of DNA from this species in rat feces, but only during the period of capsule intake.     

The effect of protein concentration and heat treatment temperature on micellar casein–soy protein mixtures

The objective of this study was to investigate the effect of concentration and temperature on the rheological properties of soy proteins (SP) and micellar casein (MCN) systems. Individual and mixed (1:1) protein systems of 2–15% concentration were prepared and heat treated for 5 min at 40–90 °C. After cooling to 20 °C, their rheological properties were determined using steady-shear rheology. Zeta potential and particle size measurements were also conducted. Both proteins were negatively charged under all experimental conditions, but the absolute values of zeta potential and thus the stability of the protein solutions decreased with temperature and concentration. For SP solutions, viscosity and apparent yield stress increased with concentration. Shear thinning behavior was prevalent, becoming more pronounced with increasing concentration. Heat treatments at T ≥ 80 °C induced glycinin denaturation, followed by aggregation and network formation when C ≥ 7.5%. Heat treatment did not significantly affect viscosity of MCN systems, while increasing concentration resulted in a significant increase in apparent viscosity and apparent yield stress. Most MCN systems exhibited Newtonian flow behavior, with the exception of systems with C ≥ 12.5% treated at T ≥ 80 °C, which became slightly shear thickening. Mixed SP–MCN systems mimicked the behavior of SP, with most values of rheological parameters intermediate between SP and MCN-only systems. Mixtures of 7.5–12.5% concentration treated at 90 °C displayed local phase separation, low viscosity and apparent yield stress, while 15% mixtures treated at 90 °C showed protein aggregation and incipient network formation. The data generated in this study can be used to develop a range of protein based products with unique flow characteristics and storage stability.     

Characterization of quinoa protein–chitosan blend edible films

Quinoa protein/chitosan films were obtained by solution casting of blends of quinoa protein extract (PE) and chitosan (CH). Films from a PE/CH blend were characterized by FTIR, X-ray diffraction, thermal analysis, and SEM. The tensile mechanical, barrier, and sorption properties of the films were also evaluated. The blend of PE with CH yielded mechanically resistant films without the use of a plasticizer. The film had large elongation at break, and its water barrier properties showed that they were more hydrophilic than CH film. The thickness and water-vapor permeability of PE/CH (v/v) 1/1 blend film increased significantly compared to pure CH films. CH films are translucent in appearance and yellowish in blend with PE. By blending anionic PE with cationic CH an interaction between biopolymers was established with different physicochemical properties from those of pure CH. Drying and sorption properties show significant differences between PE/CH blend film and CH film. The structural properties determined by XRD, FTIR and TGA showed a clear interaction between quinoa proteins and CH, forming a new material with enhanced mechanical properties.     

Effect of κ-carrageenan on milk protein polysaccharide mixtures

The effect of κ-carrageenan (0, 0.025, 0.05%) on phase separation between polysaccharides (0.36% of locust bean gum (LBG), guar gum, or xanthan gum) and milk proteins (from 10.5% skim milk powder) in solution was studied. Xanthan gum was seen to be the most incompatible with milk proteins, followed by guar gum and LBG. Casein micelles were more incompatible with all polysaccharides than whey proteins. Whereas at either concentration κ-carrageenan inhibited visual phase separation, it was seen by transmission electron microscopy that samples with κ-carrageenan showed microscopic phase separation. Samples with 0.05% κ-carrageenan and either LBG or guar gum and all samples with xanthan gum could be described rheologically as weak gels, while those with no or 0.025% κ-carrageenan and either LBG or guar gum could be described as concentrated solutions. Thus, no correlation was seen between the inhibition of macroscopic phase separation by κ-carrageenan and the formation of a weak gel in solution.     

Aggregation kinetics and ��-potential of soy protein during fractionation

The mechanism of soy protein fractionation was explored. A Focused Beam Reflectance Measurement (FBRM) technique was used to study the effects of Ca²⁺ and Mg²⁺ concentrations on the aggregation/precipitation processes of soy globulins. An electrophoretic technique was used to provide information about the electrical charge of the resulting individual protein precipitate. FBRM measurements demonstrated the presence of a two-step process for glycinin (around 90% purity) aggregation. First, in the absence of Ca²⁺ and Mg²⁺, about 9730 counts of the primary particles (~ 3.33 ??m) per second were formed immediately, with the pH being adjusted to 5.8 for glycinin precipitation. Second, more than 90% of the primary particles aggregated slowly within approximately 15 min into larger secondary particles (~ 54.72 ??m). The addition of Ca²⁺ and Mg²⁺ significantly increased the amount of the primary aggregate of soy globulins and altered their aggregation process. However, the number of primary particles induced by Ca²⁺ was smaller than that by Mg²⁺. The aggregation kinetic pattern for soy globulins in the second precipitation step (mainly ??-conglycinin) significantly differed from that of the first precipitation step (mainly glycinin) in the stability of secondary particles. Electrophoretic measurements showed that, as Ca²⁺ and Mg²⁺ concentrations were increased, the net ??-potential of the glycinin-rich fractions decreased, and the reduced effect by using Ca²⁺ was greater than that by using Mg²⁺. The different influences on the ??-potential for soy protein between Ca²⁺ and Mg²⁺ were probably due to their different influences on phytate precipitation, respectively. The combined effects of Ca²⁺ and Mg²⁺ and phytate led to the reduction in the ??-potential and thus to colloidal stability for soy globulins, resulting in more protein precipitation.     

Properties of konjac glucomannan–whey protein isolate blend films

Edible composite packaging has been developed by blending biocomponents for specific applications, aiming to take advantage of complementary functional properties or to overcome their respective flaws. The aim of this work was to study the effect of incorporation of whey protein isolate (WPI) on the properties of konjac glucomannan (KGM) based films. Five aqueous solutions of KGM and/or WPI were prepared by casting and solvent evaporation of 1:0, 0.8:3.4, 0.6:3.6, 0.4:3.8 and 0:4.2 g KGM:g WPI/100 g solution. Glycerol (Gly) was used as a plasticizer at 1.5 and 1.8 g/100 g solution. The result showed that incorporated WPI proportionally increased transparency of KGM-based films. An increase in proportion of WPI resulted in decreased tensile strength and elastic modulus as well as improved flexibility. The incorporation of WPI into the KGM matrix led to an increase in water insolubility which enhanced product integrity and water resistance. Nevertheless, WPI did not improve water vapor barrier of KGM–WPI films. WPI and blend film with the highest concentration of WPI could be heat sealed at 175 °C. Overall, the range of Gly in this study did not apparently affect properties of the films.     

Analytical techniques for the study of polyphenol–protein interactions

This mini review focuses on advances in biophysical techniques to study polyphenol interactions with proteins. Polyphenols have many beneficial pharmacological properties, as a result of which they have been the subject of intensive studies. The most conventional techniques described here can be divided into three groups: (i) methods used for screening (in-situ methods); (ii) methods used to gain insight into the mechanisms of polyphenol–protein interactions; and (iii) methods used to study protein aggregation and precipitation. All of these methods used to study polyphenol–protein interactions are based on modifications to the physicochemical properties of the polyphenols or proteins after binding/complex formation in solution. To date, numerous review articles have been published in the field of polyphenols. This review will give a brief insight in computational methods and biosensors and cell-based methods, spectroscopic methods including fluorescence emission, UV-vis adsorption, circular dichroism, Fourier transform infrared and mass spectrometry, nuclear magnetic resonance, X-ray diffraction, and light scattering techniques including small-angle X-ray scattering and small-angle neutron scattering, and calorimetric techniques (isothermal titration calorimetry and differential scanning calorimetry), microscopy, the techniques which have been successfully used for polyphenol–protein interactions. At the end the new methods based on single molecule detection with high potential to study polyphenol–protein interactions will be presented. The advantages and disadvantages of each technique will be discussed as well as the thermodynamic, kinetic or structural parameters, which can be obtained. The other relevant biophysical experimental techniques that have proven to be valuable, such electrochemical methods, hydrodynamic techniques and chromatographic techniques will not be described here.     

Effect of protein–polysaccharide mixtures on the continuous manufacturing of foamed food products

The aim of this work is to understand the effects of protein and polysaccharide interactions on the physicochemical properties of highly viscous Newtonian model foods and their impact on continuous foaming operation in laminar flow conditions. Model foods consisted of modified glucose syrups. Foaming was carried out at constant gas-to-liquid flow rate ratio as a function of rotation speed. Overrun, mean bubble diameter d₃₂ and stability over time were used to characterize foams. Results showed that blow-by occurred during foaming of models including either 0.1% guar or xanthan without proteins, while 0.1% pectin allowed a total incorporation of the gas phase with large bubbles. For proteins, models with 2% whey protein isolate (WPI) were able to form foams with the desired overrun and small bubbles, while foaming was less effective with 2% Na-caseinates. With WPI, guar addition did not improve significantly foam properties. Overrun was reduced in WPI–xanthan mixtures, probably because the matrix exhibited viscoelastic trends even though xanthan decreased d₃₂. WPI–pectin mixtures provided abundant and stable foams with the smallest d₃₂ and the best stability because WPI reinforced the time-dependent behaviour of pectin recipes. However, blow-by was observed with 0.1% pectin when WPI was replaced by Na-caseinates, which demonstrates the key role of specific protein–polysaccharide interactions on overrun. Conversely, bubble diameters in foams were governed by process parameters and could be adequately described using a laminar Weber number based on foam viscosity measured during foaming for all model foods that provided stable foams.     

Effects of protein interactions on properties and microstructure of zein–gliadin composite films

Zein and gliadin are both readily dissolved in aqueous ethanol and have good film-forming property. This article describes an attempt to improve the flexibility of zein films by the addition of gliadin to the zein film-forming solution. The properties of zein–gliadin composite films, i.e., color, transparency, moisture content, water solubility, water vapor permeability, dynamic contact angle which in turn affected the mechanical property, water resistance and glass transition temperature of films were investigated. The contents of second structure were characterized via Fourier transform infrared spectroscopy (FTIR), whereas morphology of films was examined by scanning electron microscopy (SEM). It was observed that the addition of gliadin enhanced the strain at break of zein–gliadin composite films as a result of the increase in the content of α-helix, β-turn structures and decrease in the level of β-sheet structure. The water resistance of films decreased with the content of gliadin increasing. Morphology of composite films showed that gliadin and zein organized a homogeneous material. This work opens a new perspective for zein in flexible food package.     

Influence of sugars on the characteristics of glucono-δ-lactone-induced soy protein isolate gels

The effects of the reducing sugars (glucose and lactose) and the non-reducing sugar (sucrose), heated in combination with soy protein isolate (SPI) at neutral pH, on the physicochemical and rheological properties of SPI were determined. After formation of gels induced by glucono-δ-lactone (GDL), the textural profile and physicochemical bonds of the non-heated and heated SPI gels were investigated. The gelation of SPI was induced in three stages of processing that is similar to some tofu-making procedures. First, SPI was heated in the presence of sugars at neutral pH above the denaturation temperature of SPI; then gelation was induced by GDL at iso-electric pH and finally the acidic gels were heat treated again. Heat treatment with glucose at neutral pH resulted in SPI with higher glycation degree than with lactose, whereas SPI heat treated in the presence of sucrose was not glycated. GDL-induced gels of SPI glycated with glucose was more soluble in water than gels of SPI reacted with lactose, which in turn was more soluble than the control and gels of SPI heated in the presence of sucrose. This indicates a change in the net charge of proteins caused by the glycation reaction. Glucose and lactose had a protective effect on protein denaturation at neutral pH, albeit less than sucrose, resulting in GDL-induced gels with increased water holding capacity and reduced gel hardness than sucrose. Chemical analysis indicated that disulphide bonds were involved in maintaining the structure of the gels, and solubility profiles of gels in different buffers indicate that other types of covalent bonds besides disulphide bonds were formed in gels of glycated SPI, resulting in reduced gel elasticity.     

Taste modification of amino acids and protein hydrolysate by α-cyclodextrin

The objective of this work was to characterize the formation of amino acid inclusion complexes with α-cyclodextrin (α-CD) and evaluate the influence of added α-CD on the taste perception of amino acids and hydrolyzed soy protein at pH 4.5. The formation of the inclusion complexes of phenylalanine, tryptophane, tyrosine, isoleucine, proline and histidine with α-CD were detected by nuclear magnetic resonance techniques (ROESY and DOSY) and the sensory characteristics of soy hydrolysates were judged by a panel of trained tasters. It was concluded that these amino acids form inclusion complexes with α-CD and the order of affinity for the α-CD cavity is phenylalanine ≈ tryptophane > proline > isoleucine ≈ tyrosine ≈ histidine. α-CD alters the bitter taste perception of the amino acids and reduces the bitter taste from hydrolyzed soy protein. These results indicate a potential use of α-CD for debittering protein hydrolysates in acidic beverages.     

Size of native and heated casein micelles,content of protein and minerals in milk from Norwegian Red Cattle—effect of milk protein polymorphism and different feeding regimes

Milk samples of 59 cows of the Norwegian Red Cattle breed receiving three different supplementary concentrates, were analysed for genotypes of caseins and whey proteins, the content of different milk salts (Ca²⁺, Ca, Mg and citrate), the content of total protein, casein and whey protein and the mean micellar size of native and heated casein micelles. The genotype of α_s1-casein had a statistically significant effect on the content of protein and casein, and the content of whey protein and the casein number were significantly influenced by different feeding regimes, and the content of citrate. The mean size of native and heated casein micelles was significantly influenced by the feeding regimes, genotype of α_s1-casein (native mean size only) and κ-casein, pH and the content of casein, whey protein and casein number. The heat-induced changes in mean micellar size were significantly affected by the calcium ion activity which accounted for approximately 40% of the total variation.     

Starch–protein interaction in the rumen of weaned dairy calves

The objectives of this study were to investigate the effect of starch and protein interaction on rumen environment, in situ digestion, and total-tract digestibility of nutrients in weaned dairy calves between 8 and 16 wk of age. Sixteen rumen-cannulated calves were randomly divided into 4 dietary treatment groups with 4 calves fed in each treatment. The treatment diets had 2 levels of starch [18%, low starch (LS), or 38%, high starch (HS)] and 2 levels of protein [16%, low protein (LP), or 22%, high protein (HP)] on a dry matter (DM) basis in calf grower: (1) LPLS, (2) LPHS, (3) HPLS, and (4) HPHS. Calves were fed for ad libitum intake (95% assigned grower and 5% grass hay), and refusals were collected weekly. Total-tract digestibility collection and in situ digestibility procedures were performed for each calf at 11 and 15 wk. Samples for in situ digestibility, grass hay (GH), soybean hulls (SBH), wheat middlings (WM), ground corn (GrC), and soybean meal (SBM) were incubated for 9 and 24 h. There was no starch and protein interaction on total-tract digestibility of calves. Total-tract DM, neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibility, and feed efficiency were affected by both protein and starch inclusion level in calf diet. Total-tract starch digestibility was lower for LS diets. Dry matter digestibility and feed efficiency were greater in calves fed HP and HS diets compared with calves fed LP and LS diets, respectively. Fiber digestibility (NDF and ADF) was less in calves fed HS diets compared with calves fed LS diets but was greater in calves fed HP diets compared with calves fed LP diets. Level of protein did not affect in situ DM and NDF disappearance of GH, but HP increased in situ DM and NDF disappearance of SBH. High-starch diets decreased DM and NDF disappearance of both GH and SBH. At 20 h after feeding, ruminal pH was 0.51 unit higher in calves fed HPHS compared with calves fed LPHS. Total ruminal VFA and proportion of propionate was greater with HS versus LS, whereas proportion of acetate was greater with LS versus HS. The DM disappearance of SBM and WM and NDF disappearance of WM was greater for calves fed HPHS compared with calves fed LPHS at 11 wk of age. In our study, when HP was fed with HS, rumen pH, in situ digestion of WM and SBM, and total-tract digestion of DM, NDF, and ADF increased. This provides evidence for starch–protein interaction in the rumen of recently weaned dairy calves. Improvements in total-tract and in situ digestibility suggest that both protein and starch levels are important for 8- to 16-wk-old calves.     

Development of milk protein fiber/ wool blended yarn北大核心

The blended yarn was produced by 15% milk protein fiber and 85% wool. To ensure the dyeing effect, the milk protein fiber and wool were dyeing with soft water respectively. Before spinning, it should pretreat the milk protein fiber with 1% (owf) antistantic agent and 0. 9% (owf) hair oil, to match the moisture at 13% -14%. Gilling put 6 word needle comb and the antistatic coating rubber roller, which speed was low. Roving compaction should slant small, while roving areas after the distance should be appropriately amplification.