The effect of ionic strength on the rheology of pH-induced bovine serum albumin/κ-carrageenan coacervates

Protein/polysaccharide coacervates are frequently applied to food products to control the rheology. This study investigated the effect of ionic strength (I) on the rheology of pH-induced protein/polysaccharide coacervates. Bovine serum albumin (BSA) and κ-carrageenan were used as a model protein and a polysaccharide, respectively. As the formation of BSA/κ-carrageenan coacervates increased the turbidity of an aqueous mixture, pH_c, pH_?, and pH_max values were identified corresponding to the pH of the formation of soluble coacervates, insoluble coacervates, and large insoluble coacervates respectively. Based on pH_c, pH_?, and pH_max, a state diagram of BSA/κ-carrageenan coacervation versus pH and I was constructed. Involvement of salt in coacervation screened out the electrostatic interaction between BSA and κ-carrageenan coacervation, resulting in the shift of pH_c, pH_?, and pH_max to lower pH. The shift was linearly changed to 1/I^1/2 that corresponded to the Debye length. BSA/κ-carrageenan coacervates were more elastic than viscous. The transition from insoluble coacervates to large insoluble coacervates contributed to enhancement of the rheology, especially in elasticity. An increase in the I of a BSA/κ-carrageenan mixture reduced the degree of coacervation and the elasticity, and the viscosity of BSA/κ-carrageenan coacervates.     

Microencapsulation of flaxseed oil in flaxseed protein and flaxseed gum complex coacervates

Flaxseed oil, a rich source of omega-3 fatty acids, was microencapsulated in a novel matrix formed by complex coacervation between flaxseed protein isolate (FPI) and flaxseed gum (FG). This matrix was crosslinking with glutaraldehyde. Liquid microcapsules with three core (oil)-to-wall ratios (1:2, 1:3 and 1:4) were prepared and spray-dried or freeze-dried to produce powders. The microencapsulation efficiency, surface oil, morphology and oxidative stability of these microcapsules were determined. The spray-dried solid microcapsules had higher oil microencapsulation efficiency, lower surface oil content, smoother surface morphology and higher oxidation stability than the freeze-dried microcapsules. The highest microencapsulation efficiency obtained in spray-dried microcapsules was 87% with a surface oil of 2.78% at core-to-wall ratio 1:4 and oil load 20%. The oxidation stability obtained from spray-dried microcapsules at core-to-wall ratio of 1:4 was nearly double that of the unencapsulated flaxseed oil.     

Complex coacervation of pea protein isolate and tragacanth gum: Comparative study with commercial polysaccharides

The ability of pea protein isolates (PPI) to form complex coacervates with tragacanth gum was investigated. The coacervate formation was structurally compared to three other PPI-polysaccharide interaction models: arabic gum and sodium alginate (known to form coacervates with PPI) and tara gum, a galactomannan. The effects of the pH and protein/polysaccharide ratio were mainly investigated using turbidity and zeta potential measurements. Regarding the pH of soluble complex formation, the pH of complex coacervates increased with the increase in protein-anionic polysaccharide mixture ratio. SEM images revealed the ability of the spray-drying process to form spherical particles of pea protein-polysaccharide complexes. The specificity of the microparticle surface was protein-dependent. FTIR analyses of coacervates showed the electrostatic interaction between the PPI and the polysaccharides. The results showed that tragacanth gum could be used as an alternative to gum arabic to form complex coacervates with PPI based on zeta potential measurements and coacervation yield studies.     

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.     

Characterisation of interactions between fish gelatin and gum arabic in aqueous solutions

The interactions between fish gelatin (FG) and gum arabic (GA) in aqueous solutions were investigated by turbidimetry, methylene blue spectrophotometry, zeta potentiometry, dynamic light scattering, protein assay, and state diagram at 40 °C and a total biopolymer concentration (C(T)) of 0.05%. FG underwent complex coacervation with GA, possibly via its conformational change, depending on pH and FG to GA ratio (FG:GA). The formation of FG-GA complexes was the most intense when pH 3.55 and FG:GA=50:50 (6.6:1 M ratio), however, the coacervate phase was found to be composed of a much higher FG fraction. The pH range of complex formation shifted to a higher pH region with increasing FG:GA. Soluble and insoluble FG-GA complexes were formed even in a pH region where both biopolymers were net-negatively charged. Varying C(T) significantly influenced not only the formation of FG-GA complexes but also the development and composition of coacervate phase.     

Simplified optimization for microcapsule preparation by complex coacervation based on the correlation between coacervates and the corresponding microcapsule

A close correlation between coacervates and the corresponding microcapsule was established using gelatin and gum arabic as a model system. Through turbidimetric titration and zeta-potential measurement, the optimum coacervates were obtained at pH 4.1 with the mixing ratio of 1:1 (w/w). Comparisons on morphology, yield and internal characteristic of heat-resistance were performed to testify the correlation between coacervates and the corresponding microcapsule. A simplified method for optimization of microcapsule preparation by complex coacervation was then proposed, in which the zeta-potential measurement helps to establish the approximate mixing ratios of wall materials; the turbidity analysis facilitates the finding of suitable pH corresponded to the highest turbidity to make the perfect microcapsule. Then, gelatin and sodium carboxymethyl cellulose was provided as a new combination of wall materials for the verification.     

Complex coacervation of pea protein isolate and alginate polysaccharides

Complex coacervation between pea protein isolate (PPI) and alginate (AL) was investigated as a function of pH (1.50–7.00) and mixing ratio (1:1–20:1 PPI:AL) by turbidimetric analysis and electrophoretic mobility during an acid titration. Conformational changes to the secondary structures during coacervation were also studied by Raman spectroscopy. Critical structure-forming events associated with the formation of soluble (pH 5.00) and insoluble (pH 2.98) complexes were found for a 1:1 PPI–AL mixture, with optimal biopolymer interactions occurring at pH 2.10 (pH_opt). As mixing ratios increased between 4:1 and 8:1, critical pHs shifted towards higher pH. Maximum coacervate formation at pH_opt occurred at a mixing ratio of 4:1. Electrophoretic mobility measurements showed a shift in net neutrality from pH 4.00 in homogenous PPI solutions, to pH 1.55 for the 1:1 mixture. As biopolymer ratios increased towards 8:1 PPI:AL, net neutrality shifted to higher pHs (～3.80). Raman spectroscopy revealed minimal complexation-induced conformational changes. Findings could aid in the design of pH-sensitive biopolymer carriers for use in functional food and bio-material applications.     

阿拉伯木聚糖与壳聚糖复凝聚物的制备及表征

研究了不同因素对壳聚糖-阿拉伯木聚糖复凝聚的影响,通过测定平衡相的浊度和复凝聚相收率确定玉米阿拉伯木聚糖（CAX）和壳聚糖（CS）间复合物形成的条件。采用红外光谱、热重分析、扫描电子显微镜和流变学特性对复凝聚物进行表征。研究结果表明,在CAX/CS配比为9:1,体系pH值为4.0,总固形物浓度为3%（m/m）,室温下反应10 min,最大复凝聚相产率达76.04%。红外光谱和热重分析结果表明,CAX/CS复凝聚物是通过CS中的-NH3+和CAX中-COO-之间的相互吸引形成的。用SEM扫描成像显示复凝聚物具有规则且分布均匀的多孔网络结构。CAX/CS复凝聚物粘弹特性取决于发生复凝聚的pH值。在pH值3.5和4.0时,复凝聚物为液体粘弹行为;在pH值5.0时,复凝聚物为固体粘弹性行为。pH值5.0下形成的复凝聚物的G’最高,说明除静电作用外,其他因素也可能对CAX/CS复凝聚物粘弹特性起重要作用。CAX/CS复凝聚物可以作为包封营养素和功能性因子的微囊化壁材。     

维生素A微胶囊化工艺的研究

研究了复凝聚法制备凝聚液、喷雾干燥法进行干燥的维生素A微胶囊化的工艺。以微胶囊包埋率为评价指标进行单因素及正交试验,得到最佳工艺条件为:pH值4.2、乳化时间20min、芯壁比3:1、反应温度45℃。     

Complex coacervates obtained from lactoferrin and gum arabic: Formation and characterization

Proteins and polysaccharides are the most frequently used hydrocolloids in the food industry, and their interaction can provide products such as complexes coacervates, which can be used as ingredients and biomaterials or in microencapsulation systems. In the present work, the interaction between lactoferrin (0.1, 0.2, 0.3, 0.5 and 1% w/w) and gum arabic (0.1% w/w) with various concentrations of NaCl (0, 0.01, 0.25, 0.3 and 0.5 mol/L) and at various pH values (from 1.0 to 12.0) was studied. The pH for the formation (higher turbidity) of the insoluble complex coacervates (pH_Ø1) varied according to the amount of NaCl used in the system (pH 3.5 to 5.3); these values are below the isoelectric point of lactoferrin (8.0), at which the protein is more positively charged, generating electrostatic binding. At a pH of approximately 2.0, this bond weakens, leading to the solubilization of precipitates, resulting in a sudden decrease in the turbidity (pH_Ø2). Samples containing a lower concentration of lactoferrin (0.1, 0.2 and 0.3% w/w) showed greater turbidity and consequently a higher formation of precipitates or aggregates. Even these samples, which contained a salt concentration of 0.3 mol/L, showed higher turbidity and displacement points of pH_Ø1 and pH_Ø2. The zeta potential and particle size values were used to study the influence of the pH, ionic strength and temperature on the interaction between the biopolymers. It was observed that the formation of macromolecules occurred between the isoelectric point of the protein (8.0) and the pKa of the polysaccharide (2.0), and a certain salt concentration (0.25 mol/L) led to larger particle sizes. It was observed that, at pH 7.0, a concentration of 0.1% gum arabic was able to stabilize the denaturation of the protein in solutions containing 0.1% lactoferrin, resulting in a constant particle size at all temperatures studied.     

Rheological and Microstructural Characteristics of Canola Protein Isolate−Chitosan Complex Coacervates

Wastage of byproducts such as canola meal is a pressing environmental concern, and canola protein isolate (CPI)?chitosan (Ch) coacervates have a good potential to utilize and convert the wastes into a high value added product. Yet so far, there is very limited rheological and microstructural information to assist in proper utilization of CPI ‐Ch complex coacervates. The rheological and microstructural properties of the complex coacervates formed from CPI and chitosan Ch at various CPI‐to‐Ch mixing ratios (1:1, 16:1, 20:1, and 30:1) and pH values (5.0, 6.0, and 7.0) were therefore investigated. These CPI?Ch complex coacervate phases were found to exhibit elastic behavior as evidenced by significantly higher elastic modulus (G?) compared to viscous modulus (G″) in all the tested ratios and pH ranges. They also exhibited shear‐thinning behavior during viscous flow. The complex coacervates formed at the optimum CPI‐to‐Ch ratio of 16:1 and pH of 6.0 demonstrated the highest G?, G″, and shear viscosity, which correlated well with the high strength of electrostatic interaction and thick‐walled, sponge‐like, less‐porous microstructure at this condition. The higher shear viscosity of the coacervate at pH 6.0 was most likely induced by stronger attractive electrostatic interactions between CPI and Ch molecules, due to the formation of a rather densely packed complex coacervate structure. Hence, it can be concluded that the microstructural observations of denser structure correlated well with the rheological findings of stronger intermolecular bonds at the optimum CPI‐to‐Ch ratio of 16:1 and pH of 6.0. The complex coacervate phase formed at a CPI‐to‐Ch ratio of 16:1 and pH of 6.0 also showed glassy consistency at low temperatures and rubbery consistency above its glass‐transition temperature. This study identified the potential for the newly developed CPI–Ch complex coacervate to be used as an encapsulating material due to its favorable strength. This would drastically reduce the wastage of byproducts, provide a solution to tackle the pressing global issue of wastage of byproducts, and bring about a more environmentally friendly paradigm.     

Complex coacervation between β-lactoglobulin and acacia gum in aqueous medium

The compatibility of β-lactoglobulin (β-lg) and acacia gum in aqueous medium was investigated as a function of the pH (3.6–5.0), the protein to polysaccharide weight ratio (50:1–1:20) and the total biopolymer concentration (0.1–5 wt%). The ternary phase diagrams obtained at low ionic strengths (0.005–10.7 mM) typically accounted for phase separation through complex coacervation. Thus a drop-shaped two-phase region was anchored in the water-rich corner. The electrostatic nature of the interactions between the two biopolymers was pointed out according to the pH dependence of the two-phase region's breadth. Following the absorbance of the mixtures at 650 nm, the influence of the protein to polysaccharide ratio was also demonstrated. Electrophoretic mobility (μ_E) measurements and chemical analyses of separated phases revealed the formation of soluble and insoluble coacervates and complexes. A remarkable value of the protein to polysaccharide weight ratio (2:1) at pH 4.2 gave the same protein to polysaccharide (Pr:Ps) ratio in the two phases after 2 days, implying that electrostatic interactions are maximum between β-lg and acacia gum. The increase of the total biopolymer concentration reduced the influence of pH and protein to polysaccharide ratio. Also, the increase of the pH close to the β-lg IEP reduced the influence of the total biopolymer concentration and Pr:Ps ratio. As the biopolymer content was increased at pH 3.6 and 4.2, the relative β-lg solubility increased probably because of the self-suppression of complex coacervation.     

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.     

鱼油(南海低值鱼)微胶囊化工艺的研究

壳聚糖、海藻酸钠为壁材,以自制鱼油为芯材,采用复凝聚法制备鱼油微胶囊产品。以产品的外形,粒径大小、产率、效率、缓释性能作为评价指标,系统地得出制备鱼油微胶囊的最佳工艺条件。结果表明,鱼油微胶囊的最佳工艺条件为:芯壁比为1:2,壁材(壳聚糖:海藻酸钠)比为2．5:1,乳化剂用量为0．1wt%,戊二醛用量为3．5ml,pH为9,反应温度为60℃,乳化搅拌速度为800r/s。     

The study of pH-dependent complexation between gelatin and gum arabic by morphology evolution and conformational transition

The coacervates of gelatin (G) and gum arabic (GA) were prepared in order to elucidate their pH-dependent complexation mechanism. Three biopolymers mixing ratios (MRs) (G/GA of 2:1, 1:1 and 1:2, w/w) were chosen to disclose their individual coacervates transition pattern for morphology and size distribution. The results showed that with pH decline, the coacervates became larger for the MR of 1:1 and 1:2; whereas, the trend went oppositely as to the MR of 2:1. Through the composition analysis of coacervates, such transition pattern was found to be consistent with the conversion rate of GA. Coacervates prepared by the MR of 2:1 were chosen to further investigate the formation mechanism at the molecular level. During the complexation process with pH decrease, G molecules experienced a conformational change from a flexible pattern to an ordered PPII helix. On the other hand, GA went through a transition from partly ordered PPII helix to relatively disordered conformation, and then converted to a more compact structure, called PPI helix. Such molecular transformation for both G and GA finally contributed to the smaller coacervates with pH decline, which coincided perfectly with the morphology evolution.     

Microencapsulation of flaxseed oil by soya proteins–gum arabic complex coacervation

Application of soya proteins (SP) in flaxseed oil microencapsulation based on complex coacervation was investigated. The effects of SP/gum arabic (GA) mixing ratio (1:2, 1:1 and 2:1) and pH (2.80, 3.15 and 3.75) on coacervate preparation were studied firstly. The highest coacervate yields (CY) were achieved at SP/GA = 1:1, pH 3.15, and SP/GA = 2:1, pH 3.75, which were 81.2 ± 2.0%, 88.1 ± 0.6%, respectively. Thereafter, the microencapsulation of flaxseed oil was detected in accordance with the results of the coacervation of SP/GA, that is the optimal condition for microencapsulation was corresponded to the condition where CY was the highest. Under the conditions of SP/GA = 1:1, pH 3.15, and SP/GA = 2:1, pH 3.75, the microencapsulation efficiency and total yield reached 81.5 ± 0.1% and 81.7 ± 0.4%, and 77.4 ± 3.7% and 86.7 ± 2.4%, respectively. Microscopic morphology revealed that the formation of a biopolymer shell around the oil droplets was achieved at specific conditions.     

复凝聚反应及其应用研究进展

复凝聚反应是指胶体溶液中的两种带不同电荷的聚电解质通过静电相互作用发生相分离而产生沉淀的一种现象,目前研究和应用得最多的复凝聚体系是蛋白质-多糖体系和多糖-多糖体系。由于参与复凝聚反应的聚电解质来源广泛、安全性高、可生物降解、反应可控性强,而且所得复合物有望获得新的功能性质,因此复凝聚反应及复凝聚相在医药、化工、食品等领域具有极其广阔的应用前景。本文对常见的复凝聚反应体系、影响复凝聚反应的因素、复凝聚相的性质、复凝聚反应及其产物复凝聚相在食品工业中的应用等进行了综述,以便为开发新型的复凝聚体系及拓展复凝聚反应的应用提供参考。     

辣椒红色素复凝聚微囊化工艺研究

目的以大豆分离蛋白(soybean protein isolate,SPI)和壳聚糖(chitosan,CH)为壁材,采用复凝聚法制备辣椒红色素(paprika red pigment,PRP)微胶囊。方法以微胶囊的包埋产率和包埋效率为指标,研究搅拌转速、复凝聚p H、温度、时间及SPI/CH比对微囊化效果的影响。结果 SPI/CH复凝聚法制备PRP微胶囊的最佳工艺为:将均匀的PRP乳状液冷却至室温,按SPI:CH=4:1(m:m)加入0.6%的CH溶液,此时固形物浓度为1.5%,用10%Na OH溶液调节混合液的p H至6.3,25℃、300 r/min条件下搅拌15 min得到微胶囊悬浮液,此时微胶囊的包埋产率为90.05%,效率为95.08%。所得微胶囊大小不均一,多以球形形式存在。结论 SPI/CH复凝聚体系可用于PRP的微囊化。     

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.     

Stability and rheology of water-in-oil-in-water multiple emulsions made with protein-polysaccharide soluble complexes

The morphology, stability, and rheological properties of water-in-oil-in-water (W₁/O/W₂) multiple emulsions (ME) stabilized by whey protein concentrate (W)-carboxymethylcellulose (C) soluble complexes (SC_W/C) were evaluated. The interaction pH values (pH_i) to generate SC_W/C were established through zeta potential and turbidity determinations. Six ME variations were prepared using a constant weight ratio (WR) between W and C of 3:1 (where maximum interaction occurred) and by varying the way in which the biopolymers were adsorbed at the interface (layer-by-layer, LL, or pre-formed complex, PF) and pH_i (3.7, 4.0 and 4.3). The ME initial volume-surface diameter (D_3.2) of the oil droplets ranged from 2.4 to 3.2 μm, which on turn contained numerous flocculated water droplets. Higher viscoelastic moduli values (G′ and G″), more pronounced shear thinning behaviour and smaller changes in droplet size with storage time were displayed by ME made with a pH_i value of 4.3, WR_3:1, and LL biopolymers adsorption technique.