Formulation of protein-polyphenol particles for applications in food systems.

Consumers are demanding healthy nutritious foods rich in protein (both plant and animal) and biologically-active phytochemicals from plants, which can help the body to sustain a stronger immune system and fight against oxidative stress. The aim of this study was to determine the conditions required to enhance the concentration of bioactive phenolics and protein in aggregate particles formed with whey or rice protein isolates and polyphenol-rich berry juices (cranberry and blueberry), and to evaluate properties associated with food structural functionality and stability. Different protein concentrations (10, 15 and 20% w/w) and mixing durations were tested in order to study their effects on particle formation. Addition of juices significantly increased the aggregation of whey proteins in suspensions at pH 4.5, resulting in a trimodal distribution of protein-polyphenol particles: sizes at 5 µm, 30 µm, and above 100 µm. However, the addition of polyphenol-rich juices did not markedly alter aggregation or particle size distribution for rice proteins, as compared to rice protein particles formed with imitation juice. The particles formed with 10% w/w whey protein and blueberry juice yielded the highest total phenolic (27 mg g-1 particle) and protein content (805 mg g-1 particle); therefore, only 25 g of protein-polyphenol particles delivered simultaneously 20 g of protein, and the equivalent blueberry polyphenols (∼0.64 g total polyphenolics) in two servings (150 g) of blueberries. Particles formed with whey protein and blueberry juice significantly improved foam stability and drainage half-life. Protein-polyphenol particles can be designed as ingredients to stabilize food structures and to increase delivery of health protective polyphenols and proteins in the diet.

Understanding and Controlling Food Protein Structure and Function in Foods: Perspectives from Experiments and Computer Simulations.

The structure and interactions of proteins play a critical role in determining the quality attributes of many foods, beverages, and pharmaceutical products. Incorporating a multiscale understanding of the structure-function relationships of proteins can provide greater insight into, and control of, the relevant processes at play. Combining data from experimental measurements, human sensory panels, and computer simulations through machine learning allows the construction of statistical models relating nanoscale properties of proteins to the physicochemical properties, physiological outcomes, and tastes of foods. This review highlights several examples of advanced computer simulations at molecular, mesoscale, and multiscale levels that shed light on the mechanisms at play in foods, thereby facilitating their control. It includes a practical simulation toolbox for those new to in silico modeling.

Viscosity drives texture perception of protein beverages more than hydrocolloid type.

Hydrocolloids are added to alter rheological properties of beverages but have other properties that can contribute to overall taste and texture perception. In this study, tapioca starch and λ-carrageenan were used to determine how hydrocolloid type, viscosity level (4-6 mPa·s, 25-30 mPa·s, and 50-60 mPa·s at 50 s-1 ), and complexity of the system (aqueous, skim milk, or whole milk) influence sensory taste and texture of fluids. All fluids were shear thinning; however, skim milk and whole milk solutions that contained carrageenan had much higher low shear viscosity and lower high shear viscosity than those with starch. There was a significant effect of viscosity level on sensory perception of consistency, creamy/oily, mouthcoating, and residual mouthcoating in aqueous, skim milk, and whole milk beverages, and a weak effect of hydrocolloid type. However, normalizing creamy/oily, paste, and mouthcoating against sensory consistency removed the effect of hydrocolloid type. Flavors (cream, cooked, cardboard, and melon/cardboard) were associated with the type of hydrocolloid and milk protein ingredient. Temporal dominance of sensations showed that samples exhibit similar temporal sensory profiles, although the addition of hydrocolloids enhanced dominance of creaminess even in samples without fat. Hydrocolloid type did not significantly influence mouthcoating or the persistence of astringency. Additionally, increasing viscosity from 3 to 74 mPa·s at 50 s-1 did not suppress perceived sweet or salty taste. The results suggest that in fluid systems with viscosity levels typically found in beverages, textural properties are determined by viscosity and independent of the type of hydrocolloid.

Morphological and masticatory performance variation of mouth behavior groups.

Food texture preference and product acceptance are hypothesized to be influenced by mouth behavior. Recent work identified four mouth behavior (MB) groups that describe most consumers in the United States: Chewers, Crunchers, Smooshers, and Suckers. While these behavioral preferences are thought to play a significant role in food selection and purchasing decisions, it is unknown how closely they relate to body and oral cavity measures as well as masticatory apparatus performance. Our objectives were twofold: to determine whether MB groups are related to (a) morphological variation in body, head, and oral cavity size and (b) masticatory apparatus performance (i.e., maximum jaw gape, maximum bite forces at the incisors and first molar). Measurements were collected following an online MB assessment (JBMB Mouth Behavior Typing Tool) where participants self-identified as one of the four types of consumers. As expected, univariate associations were observed between masticatory performance and overall body as well as oral cavity size. These relationships did not persist when assessed with multivariate methods. MB groups did not differ by body, head, or oral cavity measurements; maximum gape; or maximum bite force. Because of small sample sizes for Smooshers and Suckers-a reflection of their limited prevalence in the U.S. population-we interpret the results for these groups with caution. We can more confidently conclude based on our sample that Chewers and Crunchers do not differ in size, bite force, or maximum gape, suggesting other factors primarily drive food texture choice and preference in these individuals.

Starch-phenolic complexes are built on physical CH-π interactions and can persist after hydrothermal treatments altering hydrodynamic radius and digestibility of model starch-based foods.

Leveraging phenolic complexation to optimize starch functionality and digestibility is restrained by the obscurity of their physicochemical nature and molecular basis. To define starch-phenolic complexes under hydrothermal treatments, maize amylopectin and potato starch were complexed with caffeic acid, ferulic acid and gallic acid. Starch hydrothermal stability and digestibility were measured by differential scanning calorimeter and Englyst's method, respectively. While monosaccharide compositions and glycosidic linkages were analyzed by GC-MS, hydrodynamic radius and proton magnetic resonance of gelatinized complexes were measured by dynamic light scattering and NMR respectively. Compared with native starches, starch-phenolic complexes were not chemically modified and had modestly lower estimated glycemic indexes and significantly lower gelatinization temperatures (p < 0.05). Starch-phenolic complexes also had significantly lower levels of phenolic proton intensities and hydrodynamic radii relative to the control starch-phenolic mixtures (p < 0.05). These results suggested that phenolics may complex with starch through non-covalent CH-π bonds along α-(1 → 4) glycosidic chains.

Casein as a Modifier of Whey Protein Isolate Gel: Sensory Texture and Rheological Properties.

The objective of this study was to determine if casein could be used to adjust the structure of whey protein gels and alter targeted textural properties. Secondarily, we sought to determine if specific structural and mechanical properties were associated with sensory texture terms. Heat set gels were made from whey proteins alone or combined with casein in micellar or dispersed form at pH 6.0 and 5.5. Replacing the whey protein with casein produced a gel breakdown pattern that was more cohesive during mastication with increased moisture retention. Additionally, casein addition reduced gel strength but minimally altered recoverable energy (an indicator of elasticity). Structural breakdown patterns were shifted from brittle- to ductile-like fracture for gels containing dispersed casein at pH 5.5 or micellar casein at pH 6.0. Shifts in microstructure observed by confocal microscopy could not explain the changes in mechanical or sensory textures. The differentiating sensory attributes among treatments were adhesiveness, cohesiveness of mass, tackiness, firmness, fracturability, and deformability. Most notably, adding casein increased cohesiveness while maintaining water holding properties. Sensory texture properties could be explained by a combination of macroscopic structural changes (appearance), fracture properties, and postfracture breakdown pattern. Overall, it was demonstrated that casein can be used to alter whey protein gel structure such that sensory firmness and fracturability are decreased and cohesiveness is increased, while preventing a large increase in moisture release. PRACTICAL APPLICATION: There is a current desire to use alternative sources of protein in a variety of food applications, which requires the ability to design food structures with specific textural properties. Whey protein gels were used as a model soft solid structure with textural attributes of low cohesiveness and water release, and high firmness and fracturability. It was shown that adding casein modified the structure such that cohesiveness increased, firmness and fracturability decreased, and water holding ability was maintained. Using a second source of protein to modify a primary protein network appears to be a viable way to adjust textural properties.

Strain hardening and anisotropy in tensile fracture properties of sheared model Mozzarella cheeses.

We studied the tensile fracture properties of model Mozzarella cheeses with varying amounts of shear work input (3.3-73.7 kJ/kg). After manufacture, cheeses were elongated by manual rolling at 65°C followed by tensile testing at 21°C on dumbbell-shaped samples cut both parallel and perpendicular to the rolling direction. Strain hardening parameters were estimated from stress-strain curves using 3 different methods. Fracture stress and strain for longitudinal samples did not vary significantly with shear work input up to 26.3 kJ/kg and then decreased dramatically at 58.2 kJ/kg. Longitudinal samples with shear work input <30 kJ/kg demonstrated significant strain hardening by all 3 estimation methods. At shear work inputs <30 kJ/kg, strong anisotropy was observed in both fracture stress and strain. After a shear work input of 58.2 kJ/kg, anisotropy and strain hardening were absent. Perpendicular samples did not show strain hardening at any level of shear work input. Although the distortion of the fat drops in the cheese structure associated with the elongation could account for some of the anisotropy observed, the presence of anisotropy in the elongated nonfat samples reflected that shear work and rolling also aligned the protein structure.

Polyphenol-enriched berry extracts naturally modulate reactive proteins in model foods.

Healthy foods like polyphenol-rich berries and high quality edible proteins are in demand in today's functional food marketplace, but it can be difficult to formulate convenient food products with physiologically-relevant amounts of these ingredients and still maintain product quality. In part, this is because proteins can interact with other food ingredients and precipitate destabilizing events, which can disrupt food structure and diminish shelf life. Proteins in foods can also interact with human receptors to provoke adverse consequences such as allergies. When proteins and polyphenols were pre-aggregated into stable colloidal particles prior to use as ingredients, highly palatable food formulations (with reduced astringency of polyphenols) could be prepared, and the overall structural properties of food formulations were significantly improved. All of the nutritive and phytoactive benefits of the proteins and concentrated polyphenols remained highly bioavailable, but the protein molecules in the particle matrix did not self-aggregate into networks or react with other food ingredients. Both the drainage half-life (a marker of structural stability) and the yield stress (resistance to flow) of model foams made with the protein-polyphenol particles were increased in a dose-dependent manner. Of high significance in this complexation process, the reactive allergenic epitopes of certain proteins were effectively blunted by binding with polyphenols, attenuating the allergenicity of the food proteins. Porcine macrophages produced TNF-α proinflammatory cytokine when provoked with whey protein, but, this response was blocked completely when the cells were stimulated with particles that complexed whey protein with cinnamon-derived polyphenols. Cytokine and chemokine production characteristic of allergic reactions were blocked by the polyphenols, allowing for the potential creation of hypoallergenic protein-berry polyphenol enriched foods.

A comparison of the lubrication behavior of whey protein model foods using tribology in linear and elliptical movement.

Lubrication is an important factor in the sensory evaluation of food products. Tribology provides a theoretical framework and instrumental methods for evaluating frictional properties between two moving surfaces and the lubrication behavior of products between these surfaces. Relating frictional measurements to sensory properties detected during oral processing requires careful and pertinent choices in surface materials and testing conditions. The aims of this study were to investigate: (a) differences in lubrication behavior of a range of food textures and (b) the differences between linear and elliptical movement and added saliva to understand the contribution of food structure to friction. Six whey protein model food samples, ranging in texture from fluid to semisolid to soft solid, were analyzed using a pin on disk tribometer to determine the coefficient of friction (COF) across a range of sliding speeds. The samples were analyzed in their initial form and post-oral processing (n = 4) in both linear and elliptical movements. Elliptical movement slightly decreased coefficients of friction and extended the shape of the friction curve. Increases in test food viscosity decreased the COF but differences in viscosity were not apparent when test foods were mixed with saliva. Data correction for viscosity shifted the friction curves horizontally, indicating that lubrication had a greater impact upon friction than viscosity. This study provides initial insights for further comparison of linear and elliptical movement with a variety of sample compositions. PRACTICAL APPLICATIONS: Sensory perception of smoothness and creaminess are often major contributors to overall hedonic food liking and are a major reason why products high in fat and sugar are more highly preferred over other foods. These parameters are influenced by friction and lubrication between the tongue, palate, teeth, food products, and saliva during oral processing. Tribology provides an instrumental method to evaluate friction between moving surfaces that mimic oral surfaces and the lubrication behavior of foods. Trends in frictional measurements can be correlated with sensory ratings of the same foods to better understand why preferences exist for certain foods or food compositions and how to effectively improve the acceptability and enjoyment of healthier foods.

Protein-bound Vaccinium fruit polyphenols decrease IgE binding to peanut allergens and RBL-2H3 mast cell degranulation in vitro.

Peanut allergy is a worldwide health concern. In this study, the natural binding properties of plant-derived polyphenols to proteins was leveraged to produce stable protein-polyphenol complexes comprised of peanut proteins and cranberry (Vaccinium macrocarpon Ait.) or lowbush blueberry (Vaccinium angustifolium Ait.) pomace polyphenols. Protein-bound and free polyphenols were characterized and quantified by multistep extraction of polyphenols from protein-polyphenol complexes. Immunoblotting was performed with peanut-allergic plasma to determine peanut protein-specific IgE binding to unmodified peanut protein, or to peanut protein-polyphenol complexes. In an allergen model system, RBL-2H3 mast cells were exposed to peanut protein-polyphenol complexes and evaluated for their inhibitory activity on ionomycin-induced degranulation (ß-hexosaminidase and histamine). Among the evaluated polyphenolic compounds from protein-polyphenol complex eluates, quercetin, - in aglycone or glycosidic form - was the main phytochemical identified to be covalently bound to peanut proteins. Peanut protein-bound cranberry and blueberry polyphenols significantly decreased IgE binding to peanut proteins at p < 0.05 (38% and 31% decrease, respectively). Sensitized RBL-2H3 cells challenged with antigen and ionomycin in the presence of protein-cranberry and blueberry polyphenol complexes showed a significant (p < 0.05) reduction in histamine and ß-hexosaminidase release (histamine: 65.5% and 65.8% decrease; ß-hexosaminidase: 60.7% and 45.4% decrease, respectively). The modification of peanut proteins with cranberry or blueberry polyphenols led to the formation of peanut protein-polyphenol complexes with significantly reduced allergenic potential. Future trials are warranted to investigate the immunomodulatory mechanisms of these protein-polyphenol complexes and the role of quercetin in their hypoallergenic potential.

Cocoa and Whey Protein Differentially Affect Markers of Lipid and Glucose Metabolism and Satiety.

Food formulation with bioactive ingredients is a potential strategy to promote satiety and weight management. Whey proteins are high in leucine and are shown to decrease hunger ratings and increase satiety hormone levels; cocoa polyphenolics moderate glucose levels and slow digestion. This study examined the effects of cocoa and whey proteins on lipid and glucose metabolism and satiety in vitro and in a clinical trial. In vitro, 3T3-L1 preadipocytes were treated with 0.5-100 µg/mL cocoa polyphenolic extract (CPE) and/or 1-15 mM leucine (Leu) and assayed for lipid accumulation and leptin production. In vivo, a 6-week clinical trial consisted of nine panelists (age: 22.6 ± 1.7; BMI: 22.3 ± 2.1) consuming chocolate-protein beverages once per week, including placebo, whey protein isolate (WPI), low polyphenolic cocoa (LP), high polyphenolic cocoa (HP), LP-WPI, and HP-WPI. Measurements included blood glucose and adiponectin levels, and hunger ratings at baseline and 0.5-4.0 h following beverage consumption. At levels of 50 and 100 µg/mL, CPE significantly inhibited preadipocyte lipid accumulation by 35% and 50%, respectively, and by 22% and 36% when combined with 15 mM Leu. Leu treatment increased adipocyte leptin production by 26-37%. In the clinical trial, all beverages significantly moderated blood glucose levels 30 min postconsumption. WPI beverages elicited lowest peak glucose levels and HP levels were significantly lower than LP. The WPI and HP beverage treatments significantly increased adiponectin levels, but elicited no significant changes in hunger ratings. These trends suggest that combinations of WPI and cocoa polyphenols may improve markers of metabolic syndrome and satiety.

Designing Whey Protein-Polysaccharide Particles for Colloidal Stability.

Interactions between whey proteins and polysaccharides, in particular the formation of food-grade soluble complexes, are of interest because of potential functional and health benefits. A specific application that has not received much attention is the use of complexes for enhanced colloidal stability of protein sols, such as protein-containing beverages. In beverages, the primary goal is the formation of complexes that remain dispersed after thermal processing and extended storage. This review highlights recent progress in the area of forming whey protein-polysaccharide soluble complexes that would be appropriate for beverage applications. Research in this area indicates that soluble complexes can be formed and stabilized that are reasonably small in size and possess a large surface charge that would predict colloidal stability. Selection of specific proteins and polysaccharides can be tailored to desired conditions. The principal challenges involve overcoming restrictions on protein concentration and ensuring that protein remains bioavailable.

Formation of whey protein-polyphenol meso-structures as a natural means of creating functional particles.

Whey proteins provide structure and nutritional properties in food, while berry juices are thought to have biological activity that can impart anti-inflammatory health effects. In combination, the two could be an excellent source of necessary and supplemental nutrients as well as expand the functionality of whey proteins in food structures. The objectives of this investigation were to (1) develop an approach for particle formation between whey protein and cranberry, blackcurrant, or muscadine grape juices, (2) determine resulting particle composition and physical characteristics, and (3) evaluate properties related to food structure stability and maintenance of phytochemical bioactivity. Particles were formed by combining 20% w/w whey protein with juice containing 50, 250, or 500 µg g(-1) total phenolics, adjusting pH to 4.5, and centrifuging to collect aggregated particles. Particles had an approximate molar ratio of 9-50 proteins per polyphenol, and the ratio increased with increasing phenolic content of the juice used to create the particles. Particle size ranged from 1-100 µm at pH 4.5, compared to 10 µm particles that formed when whey protein isolate alone was precipitated at pH 4.5. Polyphenols and other juice components, such as acids and sugars appeared to be involved in particle formation. Particles improved foam stability, and the anti-inflammatory properties of entrapped polyphenols were maintained in the particles. Highly functional protein-polyphenol particles can be designed to stabilize food structures and simultaneously deliver polyphenols associated with health benefits.

Caramel as a Model System for Evaluating the Roles of Mechanical Properties and Oral Processing on Sensory Perception of Texture.

Food formulation can have a significant impact on texture perception during oral processing. We hypothesized that slight modifications to caramel formulations would significantly alter mechanical and masticatory parameters, which can be used to explain differences in texture perception. A multidisciplinary approach was applied by evaluating relationships among mechanical properties, sensory texture, and oral processing. Caramels were utilized as a highly adhesive and cohesive model system and the formulation was adjusted to generate distinct differences in sensory hardness and adhesiveness. Descriptive analysis was used to determine sensory texture, and mechanical properties were evaluated by oscillatory rheology, creep recovery, and pressure sensitive tack measurements. Oral processing was measured by determining activity of anterior temporalis and masseter muscles via electromyography and tracking jaw movement during chewing. The substitution of agar or gelatin for corn syrup at 0.6% w/w of the total formulation resulted in increased sensory hardness and decreased adhesiveness. Creep recovery and pressure sensitive tack testing were more effective at differentiating among treatments than oscillatory rheology. Hardness correlated inversely with creep compliance, and both stickiness and tooth adhesiveness correlated with pressure sensitive adhesive force. Harder samples, despite being less adhesive, were associated with increased muscle activity and jaw movement during mastication. Tooth packing, not linked with any mechanical property, correlated with altered jaw movement. The combination of material properties and oral processing parameters were able to explain all sensory texture differences in a highly adhesive food.

Food Protein Functionality--A New Model.

Proteins in foods serve dual roles as nutrients and structural building blocks. The concept of protein functionality has historically been restricted to nonnutritive functions--such as creating emulsions, foams, and gels--but this places sole emphasis on food quality considerations and potentially overlooks modifications that may also alter nutritional quality or allergenicity. A new model is proposed that addresses the function of proteins in foods based on the length scale(s) responsible for the function. Properties such as flavor binding, color, allergenicity, and digestibility are explained based on the structure of individual molecules; placing this functionality at the nano/molecular scale. At the next higher scale, applications in foods involving gelation, emulsification, and foam formation are based on how proteins form secondary structures that are seen at the nano and microlength scales, collectively called the mesoscale. The macroscale structure represents the arrangements of molecules and mesoscale structures in a food. Macroscale properties determine overall product appearance, stability, and texture. The historical approach of comparing among proteins based on forming and stabilizing specific mesoscale structures remains valid but emphasis should be on a common means for structure formation to allow for comparisons across investigations. For applications in food products, protein functionality should start with identification of functional needs across scales. Those needs are then evaluated relative to how processing and other ingredients could alter desired molecular scale properties, or proper formation of mesoscale structures. This allows for a comprehensive approach to achieving the desired function of proteins in foods.