Complexation between milk proteins and polysaccharides via electrostatic interaction: principles and applications – a review

This article details recent research conducted on the complexation between milk proteins and polysaccharides and the properties of the complexes, and the application of such relationships to the food industry. Complexation between proteins and polysaccharides through electrostatic interactions gives either soluble complexes in a stable solution or insoluble complexes, leading to phase separation. The formation and the stability of these complexes are influenced by pH, ionic strength, ratio of protein to polysaccharide, charge density of protein and polysaccharide as well as processing conditions (temperature, shearing and time). The functional properties of milk proteins, such as solubility, surface activity, conformational stability, gel‐forming ability, emulsifying properties and foaming properties, are improved through the formation of complexes with polysaccharides. These changes in the functional properties provide opportunities to create new ingredients for the food industry.     

Structure and Technofunctional Properties of Protein-Polysaccharide Complexes: A Review

Food proteins and polysaccharides are the two key structural entities in food materials. Generally, interactions between proteins and polysaccharides in aqueous media can lead to one- or two-phase systems, the latter being generally observed. In some cases of protein-polysaccharide net attraction, mainly mediated through electrostatic interactions, complex coac-ervation or associative phase separation occurs, giving rise to the formation of protein-polysac-charide complexes. Physicochemical factors such as pH, ionic strength, ratio of protein to polysaccharide, polysaccharide and protein charge, and molecular weight affect the formation and stability of such complexes. Additionally, the temperature and mechanical factors (pressure, shearing rate, and time) have an influence on phase separation and time stability of the system. The protein-polysacchaide complexes exhibit better functional properties than that of the proteins and polysaccharides alone. This improvement could be attributed to the simultaneous presence of the two biopolymers, as well as the structure of the complexes. Consequently, the interesting hydration (solubility, viscosity), structuration (aggregation, gelation) and surface (foaming, emulsifying) properties of these complexes can be used in a number of domains. Among others, these could be macromolecular purification, microencapsulation, food formulation (fat replacers, texturing agents), and synthesis of biomaterials (edible films, artificial grafts).     

Covalent interactions between proteins and oxidation products of caffeoylquinic acid (chlorogenic acid)

BACKGROUND: The interactions between phenolic compounds and proteins can modify protein properties important in the food industry. To understand the effects of these interactions, the covalent interactions between caffeoylquinic acid (chlorogenic acid, CQA) oxidised by polyphenol oxidase (PPO) at acidic pH 6 (pH 6) and α‐lactalbumin, lysozyme and bovine serum albumin (BSA) were compared with non‐enzymatically induced covalent interactions at alkaline pH (pH 9). The effects of these modifications on protein properties were examined. RESULTS: Both ways of modification seemed to result in protein modification mainly via dimeric rather than monomeric CQA quinones. These modifications led to a decrease in the number of free primary amino groups of the proteins. Modification with CQA alone induced a low degree of protein dimerisation, which also occurred through the action of PPO alone. Modification drastically reduced the solubility of lysozyme over a broad pH range, whereas that of α‐lactalbumin was strongly reduced only at pH values close to its pI. The solubility of BSA was much less affected than that of the other proteins and only at acidic pH. CONCLUSION: These results indicate some similarities between modifications at pH 6 and 9 and that both modifications clearly change the functional properties of globular proteins. Copyright © 2007 Society of Chemical Industry     

Physicochemical and functional characteristics of proteins treated by a pH-shift process: a review

Advanced structures of food proteins are maintained by many forces such as hydrophobic activities, electrostatic interaction, and disulphide bond interaction, which can affect their functional characteristics to a certain extent. Therefore, many approaches have been utilised to improve functional characteristics of food proteins. pH-shift process refers to the method that food proteins are treated under extreme alkaline or acid conditions followed by adjusting pH to isoelectric point or neutral pH. Many studies have showed that pH-shift process can significantly affect the functional characteristics of food proteins, for example emulsifying activities, forming property, solubility and water/oil adsorption ability. pH-shift process has been utilised to recover protein isolates from many sources including fish, fish by-products, meat processing products. Many researches have indicated that the functional and physicochemical characteristics of recovered protein isolates are significantly influenced by pH-shift process. In this paper, the latest studies regarding the functional and physicochemical characteristics of proteins treated or recovered through pH-shift process, and potential applications of obtained protein isolates in the production of the hydrolysates or used as a delivery system were reviewed.     

Modified Soy Proteins with Improved Foaming and Water Hydration Properties

Soy proteins were modified by alkali treatment at pH 10.0, followed by papain hydrolysis. Solubility, water hydration capacity (WHC), surface hydrophobicity, foaming and emulsifying properties of unmodified, alkali-treated, and papain-modified soy protein (PMSP) were compared. PMSP exhibited higher solubility (100% at pH > 7.0), WHC (3.13) and hydrophobicity (40.8) than unmodified soy protein which had solubility 68.5%, WHC 0.21, and hydrophobicity 8.1. The PMSP had foaming capacity (22.0 mL) similar to egg white (21.2 mL) at pH 7.0; and enhanced foam stability (36.4) compared to the unmodified control (32.9). In general, alkali-treated soy had lower functional properties. Emulsifying properties of PMSP and alkali treated soy were unchanged by the modification. PMSP could be used as an egg white substitute in foaming applications at neutral pH.     

FUNCTIONAL PROPERTIES OF HIGH HYDROSTATIC PRESSURE-TREATED WHEY PROTEIN

Surface hydrophobicity, solubility, gelation and emulsifying properties of high hydrostatic pressure (HHP)‐treated whey protein were evaluated. HHP treatment of whey protein buffer or salt solutions were performed at 690 MPa and initial ambient temperature for 5, 10, 20 or 30 min. Untreated whey protein was used as a control. The surface hydrophobicity of whey protein in 0.1 M phosphate buffers treated at pH 7.0 increased with an increase in HHP treatment time from 10 to 30 min. HHP treatments of whey protein in salt solutions at pH 7.0 for 5, 10, 20 or 30 min decreased the solubility of whey proteins. A significant correlation was observed between the surface hydrophobicity and solubility of untreated and HHP‐treated whey protein with r = ?0.946. Hardness of HHP‐induced 20, 25 or 30% whey protein gels increased with an increase in HHP treatment time from 5 to 30 min. An increase in the hardness of whey protein gels was observed as whey protein concentration increased. Whey proteins treated in phosphate buffer at pH 5.8 and 690 MPa for 5 min exhibited increased emulsifying activity. Whey proteins treated in phosphate buffer at pH 7.0 and 690 MPa for 10, 20 or 30 min exhibited decreased emulsifying activity. HHP‐treated whey proteins in phosphate buffer at pH 5.8 or 7.0 contributed to an increase in emulsion stability of model oil‐in‐water emulsions. This study demonstrates that HHP treatment of whey protein in phosphate buffer or salt solutions leads to whey protein unfolding observed as increased surface hydrophobicity. Whey proteins treated in phosphate buffers at pH 5.8 and 690 MPa for 5 min may potentially be used to enhance emulsion stability in foods such as salad dressings, sausage and processed cheese.     

Application of high pressure processing to improve the functional properties of pale, soft, and exudative (PSE)-like turkey meat

Improvement of functional and rheological properties of turkey breast meat proteins with different ultimate pHs at 24 h post-mortem (pH₂₄) was attempted using high pressure processing (up to 200 MPa for 5 min at 4 °C). Pressures of 50 and 100 MPa were found to increase the water holding capacity of low pH meat. At these pressures, higher protein surface hydrophobicity and greater exposure of sulfhydryl groups were evident. These elements may have contributed to improved water retention properties of the treated protein. The formation of a better gel network was also evident at 50 and 100 MPa as revealed by the dynamic viscoelastic behavior. Application of high pressure significantly (P < 0.05) increased total protein solubility in both low and normal pH meats. Aggregation of myofibrillar proteins increased in low pH meat at higher pressure (200 MPa) as revealed by SDS-PAGE profile.

Industrial relevance

A major concern in the poultry industry is reduced meat functionality, such as low water holding capacity (WHC) in low pH poultry meat leading to reduced yield causing economic loss in the production of further processed products. An alternative technology to reduce salt and improve water retention properties is by the application of high pressure processing (HPP) to produce healthier food products.     

Synthesis and physicochemical characterization of zinc-lactoferrin complexes

One trend of the modern world is the search for new biologically active substances based on renewable resources. Milk proteins can be a solution for such purposes as they have been known for a long time as compounds that can be used for the manufacturing of multiple food and non-food products. Thus, the goal of the work was to investigate the parameters of Zn-bovine lactoferrin (bLTF) interactions, which enables the synthesis of Zn-rich protein complexes. Zinc-bLTF complexes can be used as food additives or wound-healing agents. Methodology of the study included bLTF characterization by sodium dodecyl sulfate-PAGE, MALDI-TOF, and MALDI-TOF/TOF mass spectrometry as well Zn-bLTF interactions by attenuated total reflection-Fourier-transform infrared, Raman spectroscopy, scanning and transmission microscopy, and zeta potential measurements. The obtained results revealed that the factors that affect Zn-bLTF interactions most significantly were found to be pH and ionic strength of the solution and, in particular, the concentration of Zn²⁺. These findings imply that these factors should be considered when aiming at the synthesis of Zn-bLTF metallocomplexes.     

蛋白聚糖的结构和功能特性

蛋白质与多糖之间通过静电相互作用发生的复合物凝集或结合是形成蛋白聚糖的主要原因，象pH值、离子强度、蛋白质和多糖的浓度及比例、蛋白质和多糖的带电情况以及分子大小等理化条件会影响蛋白聚糖的形成及稳定性。同时，温度及一些物理因素（压力、剪切速率和时间）对此也有一定的影响。蛋白聚糖具有较好的水合作用（溶解性、粘度），结构特性和表面特性（起泡性、乳化性），可用于大分子物质的纯化，微胶囊材料，食用成分（脂肪替代物等）和生物材料的合成等（可食用薄膜，人造皮肤）。     

Effect of thermal treatment on the proteins of amaranth isolates

The effect of thermal treatment of proteins from Amaranthus hypochondriacus was studied. Two protein isolates were obtained from the defatted flour by water extraction at a pH of 9 (A9 isolate) and 11 (A11 isolate), followed by isoelectric precipitation at a pH of 5. Effect of thermal treatment (70 and 90 °C, during 3, 5, 10, 15 and 30 min) on A9 and A11 dispersions were analyzed by differential scanning calorimetry (DSC), polyacrylamide gel electrophoresis, UV spectrophotometry, superficial hydrophobicity and solubility in water. Thermal treatment induced the aggregate formation of high molecular mass stabilized by disulfide and non‐covalent bond. Thermal treatment at 70 °C produced a 30% denaturation in both, while at 90 °C A9 was more denatured than A11 (75% and 55% of denaturation, respectively). An increase in thermal stability was also detected by DSC in A9 treated at 90 °C. The denaturation process was accompanied at short heating times by an increase in UV absorbance and changes in superficial hydrophobicity. A decrease in water solubility (35–50%, depending on time–temperature conditions) was also observed for the A9 isolates. The results suggest that the A9 isolates, enriched in a globulin protein fraction, are more sensitive to thermal treatment than isolates A11 enriched in glutelin protein fraction. The changes shown by both isolates, indeed, could affect their functional properties and could definitely limit their use in food products. Copyright © 2007 Society of Chemical Industry     

The effect of alginates on in vitro gastric digestion of particulated whey protein

The aim of the study was to investigate how microparticulated and nanoparticulated whey proteins mixed with alginate respond to simulated in vitro gastric digestion conditions at pH 3.0. Initially, particle size distributions and zeta potential were measured in all mixtures at pH 3.0. Particle size distributions as well as SDS‐PAGE were used to investigate the rate of protein degradation by pepsin during simulated in vitro gastric digestion. The complexation of nanoparticulated and microparticulated whey protein with alginates causes formation of insoluble and soluble complexes, which can resist pepsin degradation to a different degree. These results highlight the potential of developing new food products, which can enhance satiety.     

Phytic acid interactions in food systems

Phytic acid is present in many plant systems, constituting about 1 to 5% by weight of many cereals and legumes. Concern about its presence in food arises from evidence that it decreases the bioavailability of many essential minerals by interacting with multivalent cations and/or proteins to form complexes that may be insoluble or otherwise unavailable under physiologic conditions. The precise structure of phytic acid and its salts is still a matter of controversy and lack of a good method of analysis is also a problem. It forms fairly stable chelates with almost all multivalent cations which are insoluble above pH 6 to 7, although pH, type, and concentration of cation have a tremendous influence on their solubility characteristics. In addition, at low pH and low cation concentration, phytate‐protein complexes are formed due to direct electrostatic interaction, while at pH >6 to 7, a ternary phytic acid‐mineral‐protein complex is formed which dissociates at high Na concentrations. These complexes appear to be responsible for the decreased bioavailability of the complexed minerals and are also more resistant to proteolytic digestion at low pH. Development of methods for producing low‐phytate food products must take into account the nature and extent of the interactions between phytic acid and other food components. Simple mechanical treatment, such as milling, is useful for those seeds in which phytic acid tends to be localized in specific regions. Enzyme treatment, either directly with phytase or indirectly through the action of microorganisms, such as yeast during bread‐making, is quite effective, provided pH and other environmental conditions are favorable. It is also possible to produce low‐phytate products by taking advantage of some specific interactions. For example, adjustment of pH and/or ionic strength so as to dissociate phytate‐protein complexes and then using centrifugation or ultrafiltration (UF) has been shown to be useful. Phytic acid can also influence certain functional properties, such as pH‐solubility profiles of the proteins and the cookability of the seeds.     

Interactions Among Calcium, Zinc and Phytate with Three Protein Sources

Various combinations of calcium (4.94 mmol), zinc (0.0071 mmol) and phytate (0.284 mmol) were added either to soy concentrate, casein or torula yeast to determine effects of their interaction on in vitro solubility of protein, calcium, zinc, and phytate (PA). Two Ca sources, calcium carbonate (CaCO₃) and calcium-citrate-malate complex (CCM) were used. Two pH levels, 2.0 and 5.5, were used to simulate gastrointestinal pH conditions. An increase in pH significantly reduced (P<0.01) Zn solubility in all treatments with all protein sources. The solubility of Ca and PA were significantly decreased (P< 0.01) when both components were present probably due to formation of insoluble Ca-PA complexes. At pH 5.5, with casein and yeast proteins, Zn was significantly more (P< 0.01) soluble in samples with CCM, in the absence of PA, than in those with CaCO₃.     

pH and ultrasound driven structure-function relationships of soy protein hydrolysate

The structural, thermodynamic and functional properties of soy protein hydrolysate (SPH) modified by treatment at different pH values (3, 5, and 9 and pH 7 as control) followed by ultrasound treatment (240 W, 30 min) were investigated. The treatment of SPH at alkaline pH combined with ultrasound treatment resulted in a reduction in the particle size and turbidity, enhancement in the surface negative charge and disulfide bond (SS) content, and exposure of more surface sulfhydryl (SH) groups, resulting in increased surface hydrophobicity and fluorescence intensity compared to those of the samples treated at pH 3–7. In addition, the alkaline-treated samples were more structurally stable than those treated at other pH values, having higher denaturation temperatures and enthalpies; moreover, these samples had higher solubility and emulsifying and foaming capacities. In addition, ultrasound-assisted pH treatment altered the secondary and tertiary structures of SPH by altering the covalent and non-covalent interactions, although there was no effect on the molecular weight distribution of proteins. In conclusion, ultrasound-assisted pH treatment is an effective method to improve physicochemical properties of SPH for applications in the food industry.     

Interaction of phytate with protein and minerals in a soybean–maize meal blend depends on pH and calcium addition

In order to investigate possible interactions of phytate with protein and minerals in simplified animal diets, studies were conducted on the solubility of endogenous phytate, protein and essential minerals in a soybean–maize meal blend within a physiological relevant pH range. The blend was mixed with water for 10 min and then allowed to incubate at 40 °C (30 min) after adjustment of the pH. Finally, soluble phytate, protein, zinc, manganese and iron were determined. Phytate and mineral solubility was highly influenced by pH whereas protein solubility was less affected. Addition of 5 g Ca²⁺ kg^?1 drastically reduced the solubility of phytate, zinc, manganese and iron at pH above 4.4, indicating that the formation of insoluble phytate–mineral complexes is increased in the presence of calcium. The action of pepsin increased the solubility of protein and phytate at pH below 4, indicating that insoluble phytate–protein complexes are present at low pH. Calcium had the same solubilising effect as pepsin at pH 2–4 but to a lesser degree. Copyright © 2007 Society of Chemical Industry     

不同pH条件下米谷蛋白的理化及结构特性研究

为探索溶解大米蛋白的最适pH值,扩大其应用范围,表征了不同pH(pH 3.0,4.0,7.0)条件下大米蛋白中主要成分——米谷蛋白的理化及结构性质。结果表明,与中性条件下相比,酸性条件下米谷蛋白的溶解度和结构性质发生了明显改变,pH 7.0时米谷蛋白分子结合紧密,形成庞大的分子聚集体,溶解度仅(6.24±1.25)%;而在酸性条件下,米谷蛋白逐渐分散,分子间二硫键断裂,呈现分散疏松的小分子体状态,pH 3.0时其溶解度最高,达到(72.47±2.36)%。     

The foam-enhancing properties of basic biopolymers

Protein systems containing both basic (high isoelectric point) and acidic (low isoelectric point) components exhibit markedly enhanced foaming properties, to an extent that significant amounts of lipids (up to 30% by weight) can be tolerated without detriment. The enhancement in foaming results from increased protein-protein interactions at the bubble surface, giving a more resilient film. Lipids, even at low levels, usually inhibit the foaming of proteins. Consequently, such mixed protein systems may have potential applications in the development of novel whipped foods.
The charge and structural requirements of basic proteins for foam enhancement have been identified. For practical purposes they must have an isoelectric point of 9 or greater and a molecular weight of 4000 daltons or greater.
Basic whey protein may be prepared by esterification in alcohol. This material has an isoelectric point between 9 and 9.5 and enhances the foaming of acidic proteins.
Chitosan, a basic polysaccharide obtained from fungi, crab and shrimp shells exhibits comparable or even superior foam-enhancing properties to basic proteins, particularly low molecular weight forms. The foaming properties of different chitosan, protein and lipid formulations are reviewed, and potential areas of use of basic biopolymers in food processing are discussed.     

Recent advances in protein derived bionanocomposites for food packaging applications

Abstract

This review article critically presents a comprehensive overview of the current advances in the research and development of proteins derived bionanocomposites used in food packaging applications. The recent interest in protein-based biomaterials is due to sustainability, renewability, biodegradability and low carbon footprint. The inherent drawbacks of proteins-based materials for food packaging applications are their low mechanical strength, poor thermal, barrier and inferior physicochemical properties. The nanoreinforced bio-based polymers called bionanocomposites provide an opportunity to overcome these issues and have ability to supersede non-biodegradable food packaging plastics produced from petroleum resources. So far, most studied protein derived bionanocomposites suitable for food packaging are soy protein isolates (SPI) and gelatin proteins. Layered silicates are the most promising nanofillers used to increase strength, improve heat resistance and enhance barrier properties of proteins derived materials while montmorillonites (MMT) is the most commonly used silicate nanofiller. This review emphases on the processing strategies used for proteins-based biomaterials, their mechanical and moisture barrier properties for food packaging applications. Different proteins and nanofillers that have been studied to date in proteins derived food packaging applications are also discussed in detail.     

Physical Properties of Mixed Dairy Food Proteins

Foods may contain more than one type of protein, and food formulators sometimes combine different proteins for desired synergistic textural benefits. Egg albumin, fish protein isolate, or soy protein isolate were blended with calcium caseinate or whey protein isolate and mixed in water adjusted to pH 2.5, 6.8, and 9.0 at 25 or 60°C. The effect of pH and temperature on solubility, viscosity, and the structure of the resulting gels were determined. The viscosity at the most soluble concentration at 25°C were: egg albumin (175.2 mPa.s/35 wt%), fish protein isolate (2207.4 mPa.s/30 wt%), soy protein isolate (2531.5 mPa.s/10 wt%), calcium caseinate (1115.8 mPa.s/15 wt%), and whey protein isolate (161.2 mPa.s/35%). In mixed protein systems viscosity values were reduced. The values for calcium caseinate or whey protein isolate with egg albumin, at the protein level of 15 g/100 g were: calcium caseinate/egg albumin (10:5 wt%) 535.1 mPa.s and whey protein isolate/egg albumin (10:5 wt%) 8.7 mPa.s. Microscopy imaging revealed changes in protein aggregation clusters during heating of calcium caseinate, egg albumin, and whey protein isolate. Egg albumin acted synergistically to increase viscosity, while fish protein isolate acted antagonistically to reduce viscosity. This knowledge is useful to manufacturers who may seek to enhance food texture by blending different proteins.     

Characterization of Textural Properties and Changes of Myofibrillar and Sarcoplasmic Proteins in Salame Felino During Ripening

Textural properties and changes of myofibrillar and sarcoplasmic proteins of salame Felino, a typical Italian dry-cured fermented sausage, were studied during ripening. Hardness, chewiness, and gumminess increased over the ripening period. Adhesiveness showed a gradual but significant decrease after 14 days and further at 28 days of ripening. A remarkable increase of the proteolysis index and a significant decrease of myofibrillar and sarcoplasmic protein solubility were observed. Hardness showed a significant negative correlation with myofibrillar protein solubility. Sarcoplasmic proteins appeared to be more susceptible to degradation.