Complex coacervation of soybean protein isolate and chitosan

The formation of coacervates between soybean protein isolate (SPI) and chitosan was investigated by turbidimetric analysis and coacervate yield determination as a function of pH, temperature, time, ionic strength, total biopolymer concentration (TB(conc)) and protein to polysaccharide ratio (R(SPI/Chitosan)). The interaction between SPI and chitosan yielded a sponge-like coacervate phase and the optimum conditions for their coacervation were pH 6.0-6.5, a temperature of 25 °C, and a R(SPI/Chitosan) ratio of four independently of TB(conc). NaCl inhibited the complexation between the two biopolymers. Fourier transform infrared spectroscopy (FTIR) revealed that the coacervates were formed through the electrostatic interaction between the carboxyl groups of SPI (-COO(-)) and the amine groups of chitosan (-NH(3)(+)), however hydrogen bonding was also involved in the coacervation. Differential scanning calorimetry (DSC) thermograms indicated raised denaturation temperature and network thermal stability of SPI in the coacervates due to SPI-chitosan interactions. Scanning electron microscopy (SEM) micrographs revealed that the coacervates had a porous network structure interspaced by heterogeneously sized vacuoles.     

Gelation properties of myofibrillar/pea protein mixtures induced by transglutaminase crosslinking

Gelation properties of mixtures of myofibrillar protein isolate (MPI)/pea protein isolate (PPI) were studied using a dynamic oscillatory rheometer and a texture analyzer to evaluate PPI as a possible meat product additive. The inclusion of microbial transglutaminase (MTG) increased the gel strength of MPI/PPI mixture (3% + 1%) more than it did for MPI (3%), but less than a 3% MPI, 1% soy protein isolate combination. The direct evidence of interaction between muscle and pea proteins in the form of new sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) bands was not found; however, the improvement in gel strength or gel peak force for the MPI/PPI mixture (3% + 1%) with inclusion of MTG suggested that some ? (γ-glutamyl) lysine (G-L) crosslinking occurred between muscle and pea proteins. It likely that pea protein acted as a non-gelling component and interspersed throughout the primary MPI gel network and the addition of MTG promoted partial crosslinking of MPI. Consequently, MTG is useful in improving gelation properties of heat-induced MPI/PPI gel.     

Associative phase separation involving canola protein isolate with both sulphated and carboxylated polysaccharides

Associative phase separation within admixtures of canola protein isolate (CPI) and anionic polysaccharides (alginate and ι-carrageenan) was investigated as a function of pH (1.50–7.00) and biopolymer weight mixing ratio (1:1–50:1, w/w) by turbidimetric analysis. Solubility of formed complexes was also studied vs. protein alone as a function of pH. In both CPI–polysaccharide systems, critical pH, associated with the onset of soluble and insoluble complexes, shifted to higher pHs as the mixing ratios increased to a 20:1 CPI–polysaccharide ratio, and then became constant. Complexes formed primarily through electrostatic attractive forces with secondary stabilisation by hydrogen bonding. Solubility of the CPI–alginate system was significantly enhanced relative to CPI alone or CPI–ι-carrageenan.     

Comparative study of functional properties of commercial and membrane processed yellow pea protein isolates

Functional properties of commercial and membrane processed pea protein isolates (PPI) prepared from yellow peas were investigated. Four protein isolates were prepared from yellow pea flour using water and KCl extractions at 25 °C followed by ultrafiltration and diafiltration (UF and DF) at pHs of 7.5 and 7.5 or 6 respectively. Following assessment of compositional attributes; solubility, foaming, flow and dynamic rheology, emulsification ability and heat-induced textural and rheological properties of prepared PPIs and a commercially available PPI were tested and compared. Membrane purification of proteins resulted in 28% to 68% reduction in phytic acid and enhanced, comparatively, the tested functional properties. Solubility of membrane processed PPIs, at all tested pHs, was superior and the lowest foaming stability and apparent viscosity were associated with commercial PPI. Gelling temperatures of water and KCl extracted PPIs, DF treated at pH 6, trimmed down to 75.7 ± 0.63 °C and 81.6 ± 0.55 °C in contrast to that of commercial PPI at above 90 °C. Similarly, the formation of firm gels, after 1 h heating at 90 °C, was associated with membrane processed PPIs whereas commercial PPI did not develop any gel.     

Gelation properties of salt-extracted pea protein isolate catalyzed by microbial transglutaminase cross-linking

Gelation is a fundamental functional characteristic of plant proteins. In this paper, a salt-extracted pea protein isolate (PPI) was mixed with microbial transglutaminase (MTG) to produce gels and the gelation properties were studied. When the MTG level increased, the magnitude of both the G′ and G″ moduli also increased, which means the gel strength increased. A second order polynomial equation was used to describe the relationships between the G′, G″ modulus and TG level. It was found that with increased heating and cooling rate at the same MTG level, G′ and G″ tended to decrease, resulting in a weaker gel. This was attributed to the rearrangement time of pea protein molecules; slower heating and cooling rates enabled protein molecules to have more time to rearrange and therefore form a stronger gel. At the same MTG level, higher pea protein concentration resulted in higher G′ and G″ values and a power law relationship was found between G′ and pea protein concentration or G″ and pea protein concentration. Frequency sweep data of PPI show that the MTG treatment resulted in higher G′ values and lower tan delta values, indicative of a stronger more elastic gel. The minimum gelation concentration was found to be 3% (w/v) with 10 U MTG treatment, lower than 5.5% required when no MTG was present. When compared to PPI and soy protein isolate (SPI) with and without 10 U MTG treatment, the gel strength of PPI with MTG was stronger than that of SPI with MTG treatment, whereas the opposite was true without the MTG treatment. SDS-PAGE showed that at the same pea protein concentration, higher MTG level induced more cross-linking as fainter bands were seen on the gel and there was a shift in the relative intensities of the bands in the molecular weight range of 35–100 kDa.     

Gelation properties of salt-extracted pea protein isolate induced by heat treatment: Effect of heating and cooling rate

Gel network formation of a salt-extracted pea protein isolate was studied using dynamic rheological measurements. The gelling point was dependent on heating rate and was unaffected by cooling rate. When both the heating and cooling rates were increased (from 0.5 to 4 °C/min) final G′ value decreased, indicative of decreased gel strength. During the heating phase, the storage modulus and loss modulus fluctuated below 1 Pa at almost constant values with the storage modulus smaller than the loss modulus until the gelling point was reached. The rate of cooling has a greater impact on the development of storage modulus than that of heating. Compared to the gel strength of commercial pea protein isolate (PPIc) and soy protein isolate (SPIc) at the same protein concentration, salt-extracted pea protein isolate (PPIs) was much stronger than PPIc but weaker than SPIc. Careful control of the heating and cooling rates enable maximum gel strength for heat-induced pea protein gel, thus enhancing utilisation of pea protein as an additive in meat food industry.     

Complex coacervation of chitosan and soy globulins in aqueous solution: a electrophoretic mobility and light scattering study

The effect of pHs and heating on the protein–polysaccharide complexation between the 0.5 wt% soy globulin (7S or 11S) and 0.1 wt% chitosan was studied. Electrophoretic and light scattering techniques were used to examine the electrical charge and aggregation of the individual biopolymers and complexes. At pH 3.0–6.5, 7S (or 11S) globulin in the presence of chitosan had significantly higher ζ‐potentials and lower particles size than 7S (or 11S) globulin alone did (e.g. 600–6000 nm at pH 5.5), indicating the formation of complexes. After heating 7S (or 11S)–chitosan mixtures had higher positive value of ζ‐potential. 7S (or 11S)–chitosan mixtures exhibited a significant increase in positive value of ζ‐potential and stability after heating at lower pH values (pH 3.3 instead of pH 4.5). Compared with other mixtures, at pH 2.5–6.0, the most remarkable decrease in aggregation was obtained for 11S–chitosan mixtures after heating at pH 3.3.     

Preparation of lutein microencapsulation by complex coacervation method and its physicochemical properties and stability

Although lutein possesses multiple valuable physiological functions, its application in food industry is limited due to the instability in adverse conditions. Using the complex coacervation method, the work is aimed to optimize the encapsulation process, investigate physicochemical properties of microcapsules and finally appraise the extent of stability improvement. The optimum process conditions determined by response surface analysis were as follows: concentration of wall materials 1.0%, ratio of core material to wall 1.25:1 and pH value 4.2, where the theoretical and practical encapsulation efficiency were 86.41% and 85.32% ± 0.63%. The particles had a confined distribution in the range of 0–30 μm, indicating a relatively homogeneous distribution. Moreover, the lutein in particles presented an improvement of ability against light, humidity, temperature. Especially, the retention rate of lutein incorporated in products reached 92.86% at 4 °C, 90.16% at 25 °C, 90.16% with the relative humidity of 33%, and 90.25% under the aerobic condition.     

Effects of plasticizers, pH and heating of film-forming solution on the properties of pea protein isolate films

Effects of glycerol (3-7% w/w) and sorbitol (4-8% w/w) concentration, pH (7.0, 9.0, 11.0) and heating (90 °C, 20 min) of film-forming solution (FFS) on the water vapor permeability (WVP), moisture content (MC), solubility, light transmission and transparency of pea protein isolate (PPI) films were investigated. Films plasticized with sorbitol exhibited significantly lower WVP, lower MC and higher solubility, in comparison with glycerol-plasticized films. Increasing glycerol content of the films led to increases in WVP and MC but did not affect film solubility. In contrast, increase in sorbitol content had no effect on permeability and MC but resulted in increased film solubility. Moisture sorption isotherms of PPI films suggested that the difference in WVP observed among films plasticized with glycerol and sorbitol might be due to the different hygroscopicity of these plasticizers. The pH of FFS did not have a significant effect on WVP and MC. Solubility of PPI films formed from non-heated FFS was not affected by pH, whereas solubility of films formed from heat-treated FFS generally increased when pH was increased from 7.0 to 11.0. Heating of FFS resulted in improved film transparency. All tested films were characterized by excellent ability to absorb UV radiation. Microstructural observation by scanning electron microscopy did not show differences between sorbitol- and glycerol-plasticized films.     

pH-dependent emulsifying properties of pea [Pisum sativum (L.)] proteins

Emulsifying properties of two partially purified legumin and vicilin (PL and PV) and protein isolate (PPI) from dry pea seeds at various pH values (3.0, 5.0, 7.0 and 9.0) were investigated. The tested emulsion characteristics included droplet size, flocculation and coalescence indices (FI and CI), creaming index, as well as interfacial protein adsorption. Some physicochemical properties of these proteins, e.g., free sulfhydryl and disulfide bond contents, protein solubility (PS), surface hydrophobicity (H_o) and thermal stability (and denaturation), were also characterized. The results indicated that emulsifying ability and emulsion stability of various pea proteins considerably varied with the preparation process, protein composition and pH. Overall, all the pea proteins exhibited least emulsifying ability at pH 5.0 (around isoelectric point), and concomitantly, the resultant emulsions were most unstable against coalescence and creaming. The emulsifying ability of these proteins at pH 3.0 was generally better than that at neutral or alkali pH values, and among all the three proteins, PL exhibited highest emulsifying ability at this pH. The flocculated state and size of droplets in fresh emulsions did not directly affect stability of these emulsions against flocculation and coalescence (upon 24 h of storage), and even creaming (up to 7 days). Interestingly, the PL and PV exhibited much better creaming stability than PPI, at pH deviating from the pI. The emulsifying properties of these proteins were not only related to their PS and H_o, but also associated with the protein adsorption and nature (e.g., viscoelasticity) of interfacial protein films. These results can greatly extend the knowledge for understanding the emulsifying properties of pea proteins, especially the pH dependence of emulsion characteristics.     

Molecular forces involved in heat-induced pea protein gelation: Effects of various reagents on the rheological properties of salt-extracted pea protein gels

The molecular forces involved in the gelation of heat-induced pea protein gel were studied by monitoring changes in gelation properties in the presence of different chemicals. At 0.3 M concentration, sodium thiocyanate (NaSCN) and sodium chloride (NaCl) showed more chaotropic characteristic and enhanced the gel stiffness, whereas sodium sulfate (Na₂SO₄) and sodium acetate (CH₃COONa) stabilized protein structure as noted by increasing denaturation temperatures (T_d) resulting in reduced storage moduli (G′). To determine the involvement of non-covalent bonds in pea protein gelation, guanidine hydrochloride (GuHCl), propylene glycol (PG), and urea were employed. The significant decrease in G′ of pea protein gels with the addition of 3 M GuHCl and 5 M urea indicated that hydrophobic interactions and hydrogen bonds are probably involved in pea protein gel formation. The increase in G′ with increasing PG concentration (5–20%), demonstrated hydrogen bonds and electrostatic interaction involvement. No significant influence was observed on G′ with addition of different concentrations of β-mercaptoethanol (2-ME), low levels of dithiothreitol (DTT), and up to 25 mM N-ethylmaleimide (NEM), which indicated that disulfide bonds are not required for gel formation, but data at higher DTT and NEM concentrations and slow cooling rates showed a minor contribution by disulfide bonds. Reheating and recooling demonstrated that gel strengthening during the cooling phase was thermally reversible but not all the hydrogen bonds disrupted in the reheating stage were recovered when recooled.     

Formation and functionality of whey protein isolate–(kappa-, iota-, and lambda-type) carrageenan electrostatic complexes

The formation of electrostatic complexes between whey protein isolate (WPI) and (κ-, ι-, λ-type) carrageenan (CG) was investigated by turbidimetric measurements as a function of pH (1.5–7.0), biopolymer weight-mixing ratio (1:1–75:1 WPI:CG) and NaCl addition (0–500 mM) to better elucidate underlying mechanisms of interaction. Emulsion stabilizing effects of formed complexes was also studied to assess their potential as emulsifiers. Complex formation followed two pH-dependent structure-forming events associated with the formation of soluble (pH_c) and insoluble (pH_?1) complexes. For both the WPI–κ-CG and WPI–ι-CG mixtures, pH_c and pH_?1 occurred at pH 5.5 and 5.3, respectively, whereas in the WPI–λ-CG mixture values were slightly higher (pH_c = 5.7; pH_?1 = 5.5). In all mixtures, maximum turbidity was found to occur near pH 4.5, before declining at lower pHs. Biopolymer mixing ratios corresponding to maximum OD was found to occur at the 12:1 ratio for both the WPI–κ-CG and WPI–λ-CG mixtures, and 20:1 ratio for WPI–ι-CG mixture. The addition of NaCl disrupted complexation within WPI–κ-CG mixtures as levels were raised, whereas when ι-CG and λ-CG was present, complexation was enhanced up to a critical Na⁺ concentration before declining. Adsorption of CG chains to the small WPI–WPI aggregates during complexation was proposed to be related to both the linear charge density and conformation of the CG molecules involved. Emulsion stability in the mixed systems (12:1 mixing ratio), regardless of the CG type (κ, ι, λ), was significantly higher than individual WPI solutions indicating enhanced ability to stabilize the oil-in-water interface.     

Characterization of a chitosan sample extracted from Brazilian shrimps and its application to obtain insoluble complexes with a commercial whey protein isolate

The rheological behaviour of chitosan solutions in 250 mM acetate buffer was studied at different pHs (25 °C). The intrinsic viscosity decreased from ∼17 dL/g to ∼14 dL/g when the pH increased from 4.7 to 6.0. Concentrated solutions (0.5–3.0% w/w) exhibited a shear-thinning behaviour which increased with increasing chitosan concentration and decreasing pH. A good fitting of the experimental data to the Cross and Carreau flow models was obtained. The elasticity of the solutions decreased with increasing pH and decreasing chitosan concentration, as a consequence of increased chain flexibility.     

Functional properties of protein fractions obtained from commercial yellow field pea (Pisum sativum L.) seed protein isolate

Commercial pea protein isolate was separated into water-soluble (WS), salt-soluble (SS), alkaline-soluble (AS) and ethanol-soluble (ES) fractions. AS fraction was the most abundant, constituting about 87% of the proteins in PPI followed by WS, SS and ES fractions in decreasing order. ES fraction consistently formed emulsions with a narrow range of smaller oil droplet sizes (0.6–19 μm) at pH 4.0, 7.0 or 9.0 compared to a wider range of sizes for emulsions stabilised by WS, SS and AS fractions. Emulsions formed with ES fraction were also the most stable (p < 0.05) over the 3 h test period at all the pH values used in this work. The WS fraction had significantly highest (p < 0.05) protein solubility and foaming capacity at all the pH values when compared to solubility of PPI, SS, and ES. Except for AS and ES fractions, foaming capacities of the protein fractions were higher at pH 9.0 than at pH 4.0 or 7.0.     

Physicochemical and textural properties of heat-induced pea protein isolate gels

Chemical and thermal properties of pea protein isolates (laboratory prepared or native; PPIn and commercial; PPIc) and textural properties of heat-set gels obtained from pea protein isolates were compared with homologous soy protein isolates (laboratory prepared, or native; SPIn and commercial; SPIc). The protein banding pattern resulting from electrophoresis separation confirmed the presence of predominant storage proteins of pea and soy seeds in the respective protein products. PPIc and SPIc had lower nitrogen solubility than their native counterparts, likely due to their denaturated state which was further confirmed by the absence of distinct endotherms in these commercial materials compared to the laboratory prepared ones. Addition of NaCl at 1.0–2.0% (w/w) to PPIn and SPIn slurries increased thermal transition temperatures for both proteins.     

Effect of whey and pea protein blends on the rheological and sensory properties of protein-based systems flavoured with cocoa

Whey and pea protein combined in different proportions (100W:0P, 75W:25P, 50W:50P, 25W:75P, 0W:100P) were used to prepare protein-based systems flavoured with cocoa and containing κ-carrageenan or κ-carrageenan/xanthan gum as thickeners. Steady and dynamic shear rheological properties of samples were measured at 10 °C and sensory differences were evaluated. Protein-based systems exhibited a shear-thinning flow behaviour that was fitted to the simplified Carreau model. Samples showed different viscoelastic properties, ranging from fluid-like to weak gel behaviour. For both types of system (with and without xanthan gum) viscosity, pseudoplasticity and elasticity rose on increasing the pea protein proportion in the blend. The sample with only whey protein obeyed the Cox-Merz rule, while in the rest of the samples complex viscosity was higher than apparent viscosity. Regarding sensory properties, the protein blend ratio mainly affected sample thickness, which rose as pea protein proportion increased. However, at the same time, the chocolate flavour and sweetness decreased and the off-flavour increased.     

Functional properties of protein hydrolysates from pea (Pisum sativum,L) seeds

The aim of this study was to investigate the effects of partial enzymatic hydrolysis on functional properties of two different pea protein isolates obtained from two pea genotypes, Maja and L1. Papain and commercial protease (Streptomyces griseus protease) were used for protein modification. Solubility, emulsifying and foaming properties were estimated at four different pH values (3.0, 5.0, 7.0 and 8.0). Papain increased solubility of L1 pea protein isolate at pH 3.0, 5.0 and 8.0, emulsifying properties and foaming capacity at all pH values. Otherwise, papain increased solubility of Maja pea protein isolate only at pH 8.0. This pea protein isolate modified with both enzymes formed emulsions with improved stability at lower pH (3.0, 5.0). The commercial protease‐prepared pea protein isolates showed generally low solubility and different emulsifying and foaming properties. Proper selection of enzyme, conditions of hydrolysis and genotypes could result in production of pea protein isolates with desirable functional properties.     

Protein enrichment of biscuits: a review

Protein enrichment of biscuits aims to reach, through a product which is widely accepted among consumers, target groups which have greater needs of this nutrient and also consumers which seek products which provide more than basic nutrition. Several protein sources have been studied for this purpose and, although the use of flours in general is more common, protein concentrates, isolates, and hydrolysates may be good alternatives. Although this reformulation seems simple, it is known that small variations in the formulations can cause important effects on dough rheology, the process, and the end product (technological and sensorial characteristics).     

Pea protein exhibits a novel Pickering stabilization for oil-in-water emulsions at pH 3.0

In this paper we reported that pea protein isolate (PPI) at pH 3.0 exhibits a novel Pickering stabilization for oil-in-water emulsions. At pH 3.0, most of the proteins in PPI were present in the nanoparticle form, with the hydrodynamic diameter of 134–165 nm depending on the concentration (c; 0.25–3.0 g/100 mL). For the emulsions formed at a specific oil fraction of 0.2, increasing the c from 0.25 to 3.0 g/100 mL resulted in a considerable reduction in the emulsion size, while their creaming stability progressively increased, and especially at c values higher than 2 g/100 mL, no creaming occurred even after storage of 20 days. Confocal laser scanning microscopy observations showed that increasing the c resulted in a progressive increase in extent of droplet flocculation, and at higher c values, a network consisting of flocculated droplets could be formed. The emulsions formed at c values above 1.0 g/100 mL exhibited extraordinary stability against coalescence. The flocculated droplet network formation was closely associated with the increased amount of adsorbed proteins at the interface. The results suggest that pea proteins exhibit a good potential to act as a kind of Pickering stabilizers for oil-in-water emulsions at acidic pHs.     

Transglutaminase treatment of pea proteins: Effect on physicochemical and rheological properties of heat-induced protein gels

Rheological properties of heat-induced pea protein isolate (PPI) gels with added microbial transglutaminase (MTGase) were studied under various reaction conditions. A positive linear relationship was observed between level of MTGase used (0 to 0.7% w/w) and shear stress and shear strain of heat-set commercial pea protein isolate (PPIc) gels at 92 °C following incubation at 50 °C. Use of MTGase allowed for preparation of PPIc gels of similar strength and elasticity as commercial soy protein isolate gels and commercial meat bologna. MTGase treatment did not alter thermal properties of PPI gels. The shear stress and strain of PPIc gels were also improved following low temperature (4 °C) incubation of PPI with MTGase. Enhancement of shear strain or gel elasticity of heat-induced PPI gels with MTGase has not been reported before and provides opportunities for extending the properties of pea proteins when developing new food products.