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.     

Effect of complexation conditions on xanthan–chitosan polyelectrolyte complex gels

Polyelectrolyte hydrogels formed by xanthan gum and chitosan can be used for encapsulation and controlled release of food ingredients, cells, enzymes, and therapeutic agents. In this study, xanthan–chitosan microcapsules were formed by complex coacervation. The effects of initial polymer concentration and chitosan solution pH on the crosslinking density of xanthan–chitosan network were investigated by swelling studies and modulated differential scanning calorimetry (MDSC) analysis. The crosslinking density was found to be less dependent on chitosan solution concentration than xanthan solution concentration and chitosan pH. The capsules were completely crosslinked at all conditions studied when initial xanthan solution concentration was 1.5% (w/v). The changes in the conformation of chitosan chains as chitosan pH approaches 6.2 were found to be important in achieving capsule network structures with different crosslinking densities. These findings indicate that the parameters studied cannot be viewed as independent parameters, as their effects on the degree of swelling are interdependent.     

Interfacial rheology of soy proteins – High methoxyl pectin films

Surface aggregating soy protein-pectin solutions are used in the production of biodegradable films intended for food packaging applications. Structural properties of the surface biopolymer network influence the engineering properties of the films, such as permeability and mechanical strength. Soy protein isolates (SPI) – high methoxyl pectin (HMP) films that develop at the air–water interface were therefore investigated by a combined interface rheological and ellipsometric approach. The behavior of pure SPI interfacial layer is that of a light cross-linked polymer network with a small regime of linear viscoelasticity response. Since SPI progressively accumulate at the air–water interface, higher protein concentration in the solution does not lead automatically to higher surface coverage but due to restricted unfolding of the proteins to weaker and fluid-like films. The rheological behavior of composite SPI–HMP solutions at the air–water interface shows that the HMP addition increases the elastic interfacial modulus. The stabilizing effect in presence of the polysaccharide is attributed to a protein–polysaccharide complex formation at the interface.     

Gelling property of soy protein–gum mixtures

Gelling properties of soy protein–gum mixtures were determined by small deformation oscillation measurement and the experimental data were analyzed with blending laws of polymers. Gel strength of soy protein–carrageenan mixture was found to follow either upper or lower bounds depending on the concentration of the constituents, suggesting the occurring of phase shift. G′ of soy protein–xanthan mixed gel always followed the upper bound, indicating that soy protein was the continuous phase regardless variations of the gum concentration. Combination of soy protein with propylene glycol alginate (PGA) produced a mixed gel with high gel strength and stayed above the upper bound at all gum concentrations examined. Covalent bonds between PGA and soy protein was suggested to contribute to the rigidity. Storage modulus of the mixture of soy protein–locust bean gum (LBG) was below the lower bound at low gum concentrations due to the limited demixing process of LBG. G′ values of the mixture of soy protein and LBG–xanthan followed the lower bound but approached upper bound on reducing protein concentration, suggesting that the presence of soy protein might inhibit LBG–xanthan mixture from forming continuous networks.     

Improving the functional properties of soy glycinin by enzymatic treatment. Adsorption and foaming characteristics

In this contribution we have determined the effect of limited enzymatic hydrolysis on the interfacial (dynamics of adsorption and surface dilatational properties) and foaming (foam formation and stabilization) characteristics of a soy globulin (glycinin, fraction 11S). The degree of hydrolysis (DH=0%, 2%, and 6%), the pH of the aqueous solution (pH=5 and 7), and the protein concentration in solution (at 0.1, 0.5 and 1 wt%) were the variables studied. The temperature and the ionic strength were maintained constant at 20 °C and 0.05 M, respectively. The rate of adsorption and surface dilatational properties (surface dilatational modulus, E, and loss angle) of glycinin at the air–water interface depend on the pH and DH. The adsorption decreased drastically at pH 5.0, close to the isoelectric point of glycinin, because of the existence of a lag period and a low rate of diffusion. The interfacial characteristics of glycinin are much improved by enzymatic treatment, especially in the case of acidic aqueous solutions. Hydrolysates with a low DH have improved functional properties (mainly foaming capacity and foam stability), especially at pH close to the isoelectric point (pI), because the native protein is more difficult to convert into a film at fluid interfaces at pH≈pI. The foam capacity depends on the rate of diffusion of protein to the interface and is much improved by the enzymatic treatment. Foam stability correlates with surface pressure and, to a minor extent, with surface dilatational modulus at long-term adsorption with few exceptions.     

Influence of molecular character of chitosan on the adsorption of chitosan to oil droplet interfaces in an in vitro digestion model

The relationship between the adsorption of chitosan to oil droplet interfaces (surface adsorption) and the molecular characteristics of the chitosan (molecular weight and charge density) was examined using an in vitro digestion model. This model involved adding different concentrations (3–10 wt%) of oil droplets to a 0.1 wt% chitosan solution at pH 3 to simulate consumption of an oil-containing meal after ingestion of chitosan. The pH was then incrementally raised to investigate pH variations that occur when a food material passes through the human digestive tract at an oil concentration of 3%. The amount of chitosan adsorbed to the oil droplet interface decreased with decreasing molecular weight (MW) and with increasing degree of deacetylation (DDA): ≈132, 85, and 78 g of oil per g of chitosan for MWs of 200, 500 and 750 kDa (all 90% DDA), respectively; and, 47, 65, and 78 g of oil per g of chitosan for DDAs of 40%, 70% and 90% (all 750 kDa), respectively at pH 3. In addition, the extent of droplet aggregation and the nature of the aggregates formed (strong versus weak; large versus small) also depended on chitosan characteristics. The pH dependence of the interaction between anionic oil droplets and cationic chitosan molecules depended on both MW and DDA. Microscope images showed formation of large flocculated structures at pH > 7 except for low DDA chitosan which remained soluble at all pH levels. Our results may have important consequences for understanding the bioactivity of chitosan, and for designing functional food ingredients to reduce lipid digestion and absorption.     

Edible films made from membrane processed soy protein concentrates

Edible films were prepared from membrane processed soy protein concentrate (MSC) at various film forming solution pHs, and their mechanical, barrier, and physical properties were compared with soy protein isolate (SPI) films. As the film solution pH increased from 7 to 10, the resulted MSC films were more transparent, yellowish, and had lower oxygen permeabilities. However, tensile strength (TS), modulus of elasticity (ME) and water vapor permeabilities of MSC films were not affected by film solution pHs. The values of MSC films prepared at pH 7 were not significantly (P>0.05) different from those of SPI films prepared at alkaline solutions (pH 8-10). The uniform TS and ME values of MSC film over the wide film solution pH ranges were attributed to the higher solubility of MSC at pH 7. For the films formed at neutral film solutions (pH 7.0), MSC films showed significantly (P<0.05) higher elongation value, film solubility, and transparency compared to SPI films.     

活性红4 离子缔合分光光度法测定壳聚糖

在pH4.5 的B-R 缓冲溶液和HAc-NaAc 缓冲溶液中,活性红4 与壳聚糖的离子缔合物在574nm 波长处的吸光度与壳聚糖含量在0～4.0μg/mL 质量浓度范围内呈良好线性关系。在B-R 缓冲溶液和HAc-NaAc 缓冲溶液中本方法的回归方程分别为A=0.3434c － 0.0355,R2=0.9996 和A=0.3229c － 0.03,R2=0.9992。两种缓冲体系测得结果无显著性差异,且不受壳聚糖分子量大小的影响。以本法测定甲壳素胶囊中壳聚糖的含量,结果与该制剂的壳聚糖标示量相符。样品测定精密度为1.43%(n=6),平均回收率为101.1%。该方法具有较强的选择性,适用于较复杂样品中壳聚糖含量的测定。     

Influence of high pressure on interactions of 11S globulin Vicia faba with ι-carrageenan in bulk solution and at interfaces

The influence of ι-carrageenan (ι-CAR) on the solution, interfacial and emulsifying properties of 11S globulin Vicia faba at low ionic strength and pH 8 has been investigated before and after high-pressure processing at 200 MPa for 20 min. The total calorimetric enthalpy (ΔH) and size exclusion chromatography studies for the pure 11S indicate that there is subunit dissociation and extensive aggregation of the protein during or following treatment. Under the same treatment conditions, 1-anilinonaphthalene-8-sulphonate (ANS) data has shown increased protein surface hydrophobicity. Pressure treatment of 11S gives much lower values of the surface tension, and apparent surface shear rheology experiments show that the molecules in the film adsorbed from the pressurised 11S are much more strongly interacting than those adsorbed from the native 11S. However, emulsions prepared with pressure processed 11S give substantially bigger droplets than those made with the untreated pure protein. Addition of ι-CAR to 11S reduces the denaturation temperature (T_m), the ΔH value, and protein surface hydrophobicity. Size exclusion chromatography at low ionic strength is indicative of complex formation. Tension measurements at the air–water interface are also consistent with the presence of a complex. Emulsions made with the simple 1:0.33 mixture of 11S+ι-CAR give emulsions with smaller droplets and pressure processing of the biopolymer mixture leads to emulsions with even smaller droplets. The presence of ι-CAR at low ionic strength appears to protect the globulin against pressure-induced aggregation.     

Influence of κ-carrageenan on the properties of a protein-stabilized emulsion

We report on the influence of κ-carrageenan (κ-CAR) on the surface activity of bovine serum albumin (BSA) and on the properties of BSA-stabilized oil-in-water emulsions. Surface tension data at low ionic strength indicate an electrostatic interaction at neutral pH which becomes much stronger at pH 6. The effect of the attractive BSA–κ-CAR interaction on the state of aggregation and creaming stability of protein-stabilized emulsions (20 vol% n-tetradecane, 1.7 wt% BSA, 5 mM) has been investigated at three pH values. At pH 6 the system behaviour is interpreted in terms of bridging flocculation leading to an emulsion droplet gel network over a certain limited polysaccharide concentration range. While the trend of behaviour is qualitatively similar to that reported recently for equivalent BSA+ι-carrageenan (ι-CAR) solutions and emulsions, the BSA–κ-CAR interaction is clearly weaker than the BSA–ι-CAR interaction under similar pH and ionic strength conditions. This means that a higher polysaccharide content is required to induce flocculation in systems containing κ-CAR, and also that the resulting emulsion gel network is weaker. The behaviour is consistent with the lower density of charged sulfate groups on the κ-CAR as compared with the ι-CAR.     

Preparation and properties of rice starch–chitosan blend biodegradable film

Biodegradable blend films from rice starch–chitosan were developed by casting film-solution on leveled trays. The influence of the ratio of starch and chitosan (2:1, 1.5:1, 1:1, and 0.5:1) on the mechanical properties, water barrier properties, and miscibility of biodegradable blend films was investigated. The biodegradable blend film from rice starch–chitosan showed an increase in tensile strength (TS), water vapor permeability (WVP), lighter color and yellowness and a decreasing elongation at the break (E), and film solubility (FS) after incorporation of chitosan. The introduction of chitosan increased the crystalline peak structure of starch film; however, too high chitosan concentration yielded phase separation between starch and chitosan. The amino group band of the chitosan molecule in the FTIR spectrum shifted from 1541.15 cm⁻¹ in the chitosan film to 1621.96 cm⁻¹ in the biodegradable blend films. These results pointed out that there was a molecular miscibility between these two components. The properties of rice starch–chitosan biodegradable blend film and selected biopolymer and synthetic polymer films were compared; the results demonstrated that rice starch–chitosan biodegradable blend film had mechanical properties similar to the other chitosan films. However, the water vapor permeability of rice starch–chitosan biodegradable blend film was characterized by relatively lower water vapor permeability than chitosan films but higher than polyolefin.     

Formation and characterization of lactoferrin/pectin electrostatic complexes: Impact of composition,pH and thermal treatment

Protein–polysaccharide complexes may be used for the development of delivery systems with applications in several industries. In the present work, the interaction of lactoferrin (LF, 0.2wt%) with a High-Methoxyl pectin (0.005–0.15wt%) in aqueous solutions was studied at different pH values (2–7) and temperatures (30–90 °C) at low ionic strength. ζ-potential and light-scattering techniques were used to provide information about the electrical charge and aggregation of individual biopolymers and complexes. At pH 7, the electrical charge went from positive to negative when increasing amounts of pectin were added to the LF solution, which was attributed to the formation of an electrostatic complex. These complexes remained soluble (low turbidity) from pH 7 to 3.5, but became turbid between pH 3.5 and 2, due to charge neutralization and bridging effects. At pH 7, the stability of LF–pectin complexes to aggregation during heating was much better than LF alone. The results of this study should provide information that will facilitate the utilization of lactoferrin as a bioactive component in food systems.     

壳聚糖/纤维素硫酸钠复合膜性能的研究

为了研究以壳聚糖和纤维素硫酸钠(NaCS)为材料制备的聚电解质复合膜在药物控制释放上的应用,考察了壳聚糖相对分子质量、NaCS相对分子质量和取代度对该复合膜机械性能(强度、韧性等)和溶胀性能的影响。结果表明,壳聚糖相对分子质量、NaCS相对分子质量和取代度对壳聚糖/NaCS复合膜的机械性质和溶胀性能都有显著的影响。壳聚糖相对分子质量越高,该复合膜的断裂伸长率越大。而NaCS相对分子质量和取代度的增大则会导致该膜的溶胀率降低。此外,溶液pH值对壳聚糖/NaCS复合膜溶胀率也有明显的影响。     

Forces Involved in Soy Protein Gelation: Effects of Various Reagents on the Formation, Hardness and Solubility of Heat-Induced Gels Made from 7S, 11S, and Soy Isolate

The effects of various reagents on the formation, hardness and solubility of heat-induced gels of soybean 7S, 11S globulins and isolate were studied. Gels were formed in 30 mM Tris HCl buffer (pH 8.0) with or without reagents by heating at 80°C for 30 min. The results indicated that electrostatic interactions and disulfide bonds are involved in the formation of 11S globulin gels; mostly hydrogen bonding in 7S globulin gels and hydrogen bonding and hydrophobic interactions in soy isolate gels. Analyses of the proteins solubilized from the gels indicated that the basic subunits of 11S globulin interact with 7S globulin in soy isolate gels. The contribution of certain acidic subunits to network formation in US soy isolate gels is limited     

Interactions between soy protein isolate and xanthan in heat-induced gels: The effect of salt addition

The influence of xanthan and/or KCl addition on the properties of heat-induced soy protein isolate (SPI) gels at pH 3.0 was studied. Changes in protein solubility and subunit composition as well as in the mechanical properties, microstructure and water holding capacity of the gels were determined. The effect of KCl addition on each biopolymer solution was also investigated. The results indicated that SPI–xanthan gels prepared without KCl were mainly stabilized by non-covalent (H-bonding and hydrophobic) and SS bond interactions, whereas in gels containing KCl, electrostatic interactions were also involved in maintaining the gel structure. The β-7S subunit was probably the fraction electrostatically linked to the xanthan. The different values found for the mechanical properties after the addition of xanthan and/or KCl, were associated with the coarseness of the gel. Xanthan and KCl probably showed a synergistic effect on the Young modulus because KCl induced a conformation transition of the xanthan molecules.     

Formation of biopolymer-coated liposomes by electrostatic deposition of chitosan

ABSTRACT:  The purpose of this study was to prepare stable biopolymer-coated liposome suspensions using an electrostatic deposition method. Liposome suspensions were produced by homogenizing 1% soy lecithin in acetate buffer (0.1 M, pH 3). Cationic chitosan (Mw approximately 200 kDa) solutions were mixed with anionic liposome suspensions ( d approximately 100 and 200 nm), and the effect of phospholipid concentration, chitosan concentration, and liposome size on the properties of the particles formed was determined. The particle size and charge (ζ-potential) were measured using dynamic light scattering and particle electrophoresis. The particle charge changed from –38 mV in the absence of chitosan to +60 mV in the presence of chitosan, indicating complex formation between the anionic liposomes and cationic chitosan molecules. Below a minimum critical chitosan concentration ( c _min), large aggregates were formed that phase separated within minutes, whose origin was attributed to formation of coacervates. On the other hand, above a maximum critical chitosan concentration ( c _max), large flocs were formed that sedimented within hours, whose formation was attributed to depletion flocculation. Minimum and maximum critical chitosan concentrations depended on liposomal concentration and size. At c _min < c < c _max, chitosan-coated liposomes were formed that did not aggregate and were stable to sedimentation. Coated liposomes had better stability to aggregation than uncoated liposomes when stored at ambient temperatures for 45 d. This study indicates that chitosan can be used to form biopolymer-coated liposomes with enhanced stability over uncoated liposomes.     

Associative interactions between chitosan and soy protein fractions: Effects of pH,mixing ratio,heat treatment and ionic strength

The objective of this paper is to explore the complexation between the soy protein fractions (glycinin and β-conglycinin) and chitosan (CS) and to investigate the influence of pH, mixing ratio, heat treatment and ionic strength. Phase behavior and microstructure showed that soluble complex and coacervate were obtained in glycinin/CS and β-conglycinin/CS mixtures at specific pHs, following a nucleation and growth mechanism. Moreover, the coacervates showed higher thermal stability than protein alone. Specially, the glycinin/CS mixture displayed a gel-like network structure at pH 5.5 and 6.0, and this structure kept the mixture soluble at a long pH region. The turbidity versus ζ-potential pattern showed that, independent of protein, the self aggregation of soy protein fractions and the coacervation of glycinin/CS and β-conglycinin/CS mixtures were all obtained at charge neutralization pH, indicating that the ζ-potential is the most critical parameter to understand the stability of soy protein/chitosan mixture. This predictive parameter was less affected by mixing ratio and heating but was significantly affected by ionic strength because mixing ratio and heating only changed the equilibrium between repulsive and attractive forces in colloid system while sodium chloride destroyed the predictability of colloidal stability via shielding charged reactive sites on both biopolymers to disrupt electrostatic interactions.     

Preparation,water solubility and rheological property of the N-alkylated mono or disaccharide chitosan derivatives

N-alkylation of chitosan was performed in a mixture of methanol and 1% acetic acid containing different amounts of monosaccharides or disaccharides including glucose, galactose, glucosamine, fructose, lactose, maltose and cellobiose. All the N-alkylated chitosan derivatives with monosaccharides were insoluble in aqueous solution (pH 7), while N-alkylated chitosan derivatives with disaccharides were easily soluble in distilled water, and the N-alkylated chitosan derivatives with lactose were soluble only at high pH. The degree of substitution (DS) of the N-alkylated chitosan derivatives increased with increasing disaccharides levels and with increasing reaction time. The reduced viscosity of the N-alkylated chitosan derivatives with disaccharides decreased with increasing DS. Apparent viscosity and pseudoplasticity of the N-alkylated disaccharide containing derivative solutions generally decreased with increasing DS. Although apparent viscosities of N-alkylated chitosan derivatives with low DS decreased with increase in pH or ionic strength, changes in high DS N-alkylated chitosan derivatives with pH values or ionic strength were not marked.     

Factors influencing the production of o/w emulsions stabilized by β-lactoglobulin–pectin membranes

A primary emulsion was prepared by homogenizing 10 wt% corn oil with 90 wt% aqueous β-lactoglobulin solution (0.5 wt% β-lg, pH 3 or 7) using a two-stage high-pressure valve homogenizer. This emulsion was mixed with aqueous pectin (citrus, 59% DE) stock solution (2 wt%, pH 3 or 7) and NaCl solution to yield secondary emulsions with 5 wt% corn oil, 0.225 wt% β-lactoglobulin, 0.2 wt% pectin and 0 or 100 mM NaCl. The final pH of the emulsions was then adjusted (3–8). Primary and secondary emulsions were ultrasonically treated (30 s, 20 kHz, 40% amplitude) to disrupt any flocculated droplets. Secondary emulsions were more stable than primary emulsions at intermediate pHs. Secondary emulsions prepared at pH 7 had smaller particle diameters (0.35 to 6 μm) than those prepared at pH 3 (0.42 to 18 μm) across the whole pH range studied, and also had smaller diameters than the primary emulsions (0.35 to 14 μm). Ultrasound treatment reduced the particle diameter of both primary and secondary emulsions and lowered the rate of creaming. The presence of NaCl screened the charges and thus the electrostatic interaction between biopolymer molecules and primary emulsion droplets. Secondary emulsions were more stable to the presence of 100 mM NaCl at low pHs (3–4) than primary emulsions. This study shows that stable emulsions can be prepared by engineering their interfacial membranes using the electrostatic interaction of natural biopolymers, especially at intermediate pHs where proteins normally fail to function.     

PROTEIN-PHYTATE INTERACTION IN SOYBEANS. III. THE EFFECT OF PROTEIN-PHYTATE COMPLEXES ON ZINC BIOAVAILABILITY

A study was conducted to determine the influence of pH on the chemical form of phytate in a commercial soy isolate and its effect on zinc bioavailability by monitoring weight gain in weanling rats. Ultrafiltration studies demonstrated that the free phytate content of soy isolate solutions was greatest at pH 5.0; above this the neutralization of soy isolates resulted in the formation of a soluble proteinphytate complex. The bioavailability of zinc from diets providing protein from egg, acid and neutral soy isolates and containing various levels of added zinc was measured. The growth of rats receiving zinc from the neutral soy isolate was significantly lower than that of rats deriving zinc from either the acidic soy isolate or egg white diets. It is proposed that the reduction in bioavailability of zinc caused by the neutralization of soy protein isolate is related to the formation of a protein-zinc-phytic acid complex resistant to hydrolysis in vivo.