Influence of protein extraction and texturization on odor-active compounds of pea proteins.

BACKGROUND: The use of plant proteins as food ingredients might be limited due to the presence of foreign or 'off' flavors, which may evolve during extraction and subsequent processing. In this study, the influence of dry (TVP) and wet (WTP) texturization on characteristic volatile compounds of two different pea protein isolates was assessed using gas chromatography-mass spectrometry-olfactometry (GC-MS-O) after direct immersion stir bar sorptive extraction (DI-SBSE). RESULTS: Twenty-four odor-active compounds were found, with a prevalence of carbonyls from fat oxidation. Nine of these compounds which are also known as major (off-) flavor contributors in peas were distinctively impacted in all texturates: hexanal, nonanal, 2-undecanone, (E)-2-octenal, (E, Z)-3,5-octadiene-2-one, (E, E)-2,4-decadienal, 2-pentyl-furan, 2-pentyl-pyridine, and γ-nonalactone. For example, hexanal, a characteristic green odorant, was reduced by up to sixfold by wet texturization, from 3.29 ± 1.05% (Pea Protein I) to 0.52 ± 0.02% (Pea WTP I). Furthermore, (E,Z)-3,5-Octadiene-2-one and (E,E)-2,4-decadienal were decreased by 1.5- and 1.8-fold when Pea Protein I and Pea TVP I were compared. CONCLUSION: An overall reduction in fat oxidation products and of green and fatty odor-active compounds was observed. The results represent a first insight into the process-related modulation of pea protein (off-) flavors to broaden the applicability of pea proteins as food ingredients.

Formulating protein-based beverages for the dysphagia diets of the elderly: viscosity, protein quality, in vitro digestion, and consumers acceptability.

BACKGROUND: Dysphagia is defined as a disorder of the swallowing mechanism. The most common management of dysphagia is diet modification by thickening food and beverages. This study aimed to obtain protein-based beverages for the dysphagia diets of the elderly, corresponding to the 'honey' (III) level of dysphagia fluids according to the National Dysphagia Diet classifications, and containing 100 g kg-1 of good-quality proteins with a high rate of hydrolysis during digestion. RESULTS: Four protein formulations made from pea proteins, milk proteins, a mixture of milk and pea proteins, and milk proteins with added konjac glucomannan, were evaluated on the basis of rheological characterization and proteolysis kinetics during in vitro digestion. The mixture of milk proteins and pea proteins, and the mixture of milk proteins with added konjac glucomannan, showed typical yielding pseudoplastic fluid behavior with similar apparent viscosity but different structural characteristics. These differences were the reason for the differences in proteolysis kinetics during digestion. The mixture of milk and pea proteins showed viscous liquid behavior and was more rapidly hydrolyzed under gastrointestinal conditions than mixtures containing milk proteins and konjac glucomannan acting as a weak gel system. CONCLUSION: We presume that geriatric consumers with swallowing difficulties may benefit from 'honey'-level viscosity, protein-based beverages containing pea and milk proteins through faster proteolysis and better bioaccessibility of amino acids during digestion. © 2020 Society of Chemical Industry.

Protein gels and emulsions from mixtures of Cape hake and pea proteins.

BACKGROUND: Portioning of frozen fish generates by-products such as fish 'sawdust' and cut-offs which can be further processed into protein concentrates and isolates. The objective of the present work was to produce gels and emulsions using recovered Cape hake protein powder (HPP). In previous works, the structures of the gels produced by HPP were found to be strong, with a high rubbery character. In this work, the addition of commercial pea proteins (PPC) to HPP gels and emulsions was studied. RESULTS: Physical properties of gels and emulsions prepared with different proportions of mixtures of PPC and HPP were evaluated. In general, gels and emulsions showed high values for whiteness and, as expected, the higher content of HPP in the protein mixtures led to higher firmness values of the gels. The gel network was rapidly formed upon heating due to the fish protein macromolecules and further reinforced by the pea protein macromolecules when cooled to 5 °C. Both visco-elastic parameters, storage and loss moduli, of the produced gels increased with the HPP proportion in the protein mixtures, corresponding to more structured systems. For the emulsions, two different pH environments were studied: 3.8 and 7.0. At neutral pH a synergy was found between the vegetable and fish protein, which is not so strong when pH is lowered to 3.8, near the isoelectric point of pea proteins (pI = 4.5). This evidence was supported by the results from the texture measurements, viscosity and visco-elastic parameters. CONCLUSIONS: Gels made from Cape hake proteins showed a softer texture and were less rubbery with the addition of pea proteins. Emulsions stabilised by these mixtures showed slightly different behaviour when produced at pH 7.0 or pH 3.8.

In situ interaction of pea peptides and bile salts under in vitro digestion: Potential impact on lipolysis.

The present work evaluated how a native pea protein isolate (PPI) affects the key roles carried out by bile salts (BS) in lipid digestion by means of the in vitro static INFOGEST protocol. Two gastric residence times were evaluated (10 and 60 min), and then the peptides obtained (GPPP) were mixed with BS at physiological concentration in simulated intestinal fluid to understand how they interact with BS both at the bulk and at the interface. Both GPPP give rise to a film with a predominant viscous character that does not constitute a barrier to the penetration of BS, but interact with BS in the bulk duodenal fluid. When the peptides flushing from the stomach after the different gastric residence times undergo duodenal digestion, it was found that for the longer gastric residence time the percentage of soluble fraction in the duodenal phase, that perform synergistically with BS micelles, was twice that of the lower residence time, leading to an increase in the solubilization of oleic acid. These results finally lead to a greater extent of lipolysis of olive oil emulsions. This work demonstrates the usefulness of in vitro models as a starting point to study the influence of gastric residence time of pea protein on its interaction with BS, affecting lipolysis. Pea proteins were shown to be effective emulsifiers that synergistically perform with BS improving the release and bioaccessibility of bioactive lipids as olive oil.

Evaluation of structure, quality, physicochemical properties, and phenolics content of pea proteins: A novel strategy through the incorporation of fermentation.

The utilization of pea proteins (PPs) is limited due to their relatively low protein digestibility (∼81%) compared to animal-based proteins, such as whey. The present investigation involved the fermentation of PPs at a concentration of 1% (w/v) using 5% (w/v) water kefir for 60 h at 25°C to improve the functional properties of PPs. The results showed a significant (p < 0.05) increase in lactic acid and acetic acid production during fermentation. These findings suggest that PPs can be effectively fermented using water kefir as a starter culture for the increased protein digestibility of PPs. The PP conformation underwent modifications, including secondary and tertiary protein structure alterations. The total phenolic compounds increased throughout the fermentation, reaching around 695.32 ± 15 mg gallic acid equivalent/100 g after 24 h of fermentation. Furthermore, the fermentation process has culminated in significant (p < 0.05) changes in the surface charge and hydrophobic properties of the fermented PPs, from -38.1 to -45.73 and 362.7 to 550.2, respectively. Fermentation using water kefir is a promising technique for improving the digestibility, protein structure, and nutritional values of PPs.

Use of Pea Proteins in High-Moisture Meat Analogs: Physicochemical Properties of Raw Formulations and Their Texturization Using Extrusion.

A new form of plant-based meat, known as 'high-moisture meat analogs' (HMMAs), is captivating the market because of its ability to mimic fresh, animal muscle meat. Utilizing pea protein in the formulation of HMMAs provides unique labeling opportunities, as peas are both "non-GMO" and low allergen. However, many of the commercial pea protein isolate (PPI) types differ in functionality, causing variation in product quality. Additionally, PPI inclusion has a major impact on final product texture. To understand the collective impact of these variables, two studies were completed. The first study compared four PPI types while the second study assessed differences in PPI inclusion amount (30-60%). Both studies were performed on a Wenger TX-52 extruder, equipped with a long-barrel cooling die. Rapid-visco analysis (RVA) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated differences in protein solubility among the different PPI types. In general, lower protein solubility led to better product quality, based on visual evaluation. Cutting strength and texture profile analysis showed increasing PPI inclusion from 30-60% led to significantly higher product hardness (14,160-16,885 g) and toughness (36,690-46,195 g. s). PPI4 led to lower product toughness (26,110 and 33,725 g. s), compared to the other PPIs (44,620-60,965 g. s). Heat gelling capacity of PPI4 was also highest among PPI types, by way of least gelation concentration (LGC) and RVA. When compared against animal meat, using more PPI (50-60%) better mimicked the overall texture and firmness of beef steak and pork chops, while less PPI better represented a softer product like chicken breast. In summary, protein content and also functionality such as cold water solubility and heat gelation dictated texturization and final product quality. High cold water solubility and poor heat gelation properties led to excessive protein cross linking and thicker yet less laminated shell or surface layer. This led to lower cutting firmness and toughness, and less than desirable product texture as compared to animal meat benchmarks. On the other hand, pea proteins with less cold water solubility and higher propensity for heat gelation led to products with more laminated surface layer, and higher cutting test and texture profile analysis response. These relationships will be useful for plant-based meat manufacturers to better tailor their products and choice of ingredients.

Exploring the oil-water interfacial behaviour of pea protein-guar gum/gellan gum mixtures relating to stability and in vitro protein digestion of model pea milk system.

Polysaccharides are effective stabilisers for plant-based milks, but their interfacial adsorption behaviours in plant milk system and effects on plant protein digestibility are still unclear. This study analysed the oil-water interfacial adsorption behaviours of pea protein concentrate (PPC) and isolate (PPI) mixed with neutral guar gum and anionic gellan gum to understand the stability and in vitro protein digestion of corresponding model pea milk system. Adding guar gum or gellan gum enhanced the interfacial adsorption stability of pea proteins without compromising proteolysis during gastrointestinal digestion. Gellan gum performed better than guar gum for long-term co-stabilising the emulsion with pea proteins, due to its electrostatic repulsion with pea proteins and stronger steric hindrance induced by thicker adsorbed interfacial layers (~10 % or ~37 % higher thickness than guar gum counterparts) as detected by a quartz crystal microbalance with dissipation (QCM-D) after rinsing. The model pea milks with 0.1 % (wt%) gellan gum were stable at 4 °C for 28 days regardless of protein types. The presence of polysaccharides at the interface enabled oil droplets to be kept separate during in vitro gastrointestinal digestion, improving the final proteolysis by ~4 to ~27 %. This result is conducive to selecting polysaccharides for developing pea protein-based beverages.

Structural evolution of pea-derived albumins during pH and heat treatment studied with light and X-ray scattering.

Pea albumins are found in the side stream during the isolation of pea proteins. They are soluble at acidic pH and have functional properties which differ from their globulin counterparts. In this study, we have investigated the aggregation and structural changes occurring to pea albumins under different environmental conditions, using a combination of size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALS) and small-angle X-ray scattering (SAXS). Albumins were extracted from a dry fractionated pea protein concentrate by precipitating the globulin fraction at acidic pH. The albumins were then studied at different pH (3, 4, 4.5, 7, 7.5, and 8) values. The effect of heating at 90 °C for 1, 3, and 5 min on their structural changes was investigated using SAXS. In addition, size exclusion of the albumins showed 4 distinct populations, depending on pH and heating conditions, with two large aggregates peaks (∼250 kDa): a dimer peak (∼24 kDa) containing predominantly pea albumin 2 (PA2), and a monomer peak of a molar mass of about 12 kDa (PA1). X-ray scattering intensities as a function of q were modeled as polydisperse spheres, and their aggregation was followed as a function of heating time. Albumins was most stable at pH 3, showing no aggregation during heat treatment. While albumins at pH 7.5 and 8 showed aggregation after heating, solutions at pH 4, 4.5, and 7 already contained aggregates even before heating. This work provides new knowledge on the overall structural development of albumins under different environmental conditions, improving our ability to employ these as future ingredients in foods.

High pressure homogenization to boost the technological functionality of native pea proteins.

Pea proteins are being increasingly used for the formulation of plant-based products, but their globular structure and the presence of aggregates can affect their technological properties. In this study, the effect of high pressure homogenization (HPH) at different intensities (60 and 100 MPa) was investigated as a pre-treatment to modulate the techno-functional properties of a pea protein isolate (IP) extracted through an alkaline extraction/isoelectric precipitation process. SDS-PAGE, circular dichroism, thermal properties, total free sulfhydryl groups, antioxidant capacity and reducing properties were evaluated along with technological indices as solubility, WHC and OHC, interfacial tension and emulsifying capacity. HPH treatments were able to unfold and modify proteins structure, leading also to a change of the relative abundance of pea protein globulins (SDS-PAGE) and of the vicilin to legumin ratio. Solubility, WHC and OHC were improved, while interfacial tension and emulsifying capacity were weakly affected. However, an enhanced physical stability over time of the emulsions prepared with the 60 MPa-treated protein was found, likely as an effect of the decreased ratio between vicilin and legumin after treatment. Results of this study will contribute to deepen the effect of the HPH technology used as pre-treatment, adding useful results and expanding knowledge about the structure and techno-functional properties of native and modified pea proteins.

Topical Application of a Formulation Containing Pea Proteins and Xyloglucan in Adult Patients with Atopic Dermatitis: A Double-blind, Parallel, Randomized, Placebo-controlled, Multicenter Study.

Objective: Atopic dermatitis (AD) management requires long-term use of drugs that come with side effects. Compounds such as xyloglucan (XG) and pea proteins (PP) are emerging alternatives to corticosteroids that have shown to restore skin barrier function in preclinical studies. This double-blind, parallel, randomized, placebo-controlled clinical trial investigated the efficacy and safety of XG and PP, in adult AD patients. Methods: Fourty-two patients with AD were randomly assigned 1:1 to receive a XG+PP treatment or the vehicle without XG+PP twice/day for 14 consecutive days for assessment at baseline, Day 8 and Day 15; follow-up visit was 14 days after the end of treatment (Day 28). Efficacy was evaluated using the Scoring Atopic Dermatitis (SCORAD) index, AD severity index (ADSI) score and patient-oriented eczema measure (POEM). Safety and tolerability were monitored as the occurrence of Adverse Events (AEs). Results: At baseline, both groups exclusively included moderate/severe AD cases. At Day 8, six patients treated with XG+PP displayed complete resolution of AD, while 15 patients had mild AD. At Day 28, 16 patients no longer had eczema, whereas five patients displayed mild AD. Notably, 21 patients in the vehicle group still displayed moderate/severe AD. Conclusion: XG and PP promote rapid and long-lasting relief, supporting its use as a safe alternative to mainstay corticosteroid treatments for AD management. The study protocol has been registered in the ISRCTN registry (TN66879853).

Mechanism and application of fermentation to remove beany flavor from plant-based meat analogs: A mini review.

Over the past few decades, there has been a noticeable surge in the market of plant-based meat analogs (PBMA). Such popularity stems from their environmentally friendly production procedures as well as their positive health effects. In order to meet the market demand, it is necessary to look for plant protein processing techniques that can help them match the quality of conventional meat protein from the aspects of sensory, quality and functionality. Bean proteins are ideal options for PBMA with their easy accessibility, high nutrient-density and reasonable price. However, the high polyunsaturated lipids content of beans inevitably leads to the unpleasant beany flavor of soy protein products, which severely affects the promotion of soy protein-based PBMA. In order to solve this issue, various methods including bleaching, enzyme and fermentation etc. are developed. Among these, fermentation is widely investigated due to its high efficiency, less harm to the protein matrix, targeted performance and low budget. In addition, proper utilization of microbiome during the fermentation process not only reduces the unpleasant beany flavors, but also enhances the aroma profile of the final product. In this review, we provide a thorough and succinct overview of the mechanism underlying the formation and elimination of beany flavor with associated fermentation process. The pros and cons of typical fermentation technologies for removing beany flavors are discussed in alongside with their application scenarios. Additionally, the variations among different methods are compared in terms of the strains, fermentation condition, target functionality, matrix for application, sensory perception etc.

Pea Proteins Have Anabolic Effects Comparable to Milk Proteins on Whole Body Protein Retention and Muscle Protein Metabolism in Old Rats.

Plant proteins are attracting rising interest due to their pro-health benefits and environmental sustainability. However, little is known about the nutritional value of pea proteins when consumed by older people. Herein, we evaluated the digestibility and nutritional efficiency of pea proteins compared to casein and whey proteins in old rats. Thirty 20-month-old male Wistar rats were assigned to an isoproteic and isocaloric diet containing either casein (CAS), soluble milk protein (WHEY) or Pisane™ pea protein isolate for 16 weeks. The three proteins had a similar effect on nitrogen balance, true digestibility and net protein utilization in old rats, which means that different protein sources did not alter body composition, tissue weight, skeletal muscle protein synthesis or degradation. Muscle mitochondrial activity, inflammation status and insulin resistance were similar between the three groups. In conclusion, old rats used pea protein with the same efficiency as casein or whey proteins, due to its high digestibility and amino acid composition. Using these plant-based proteins could help older people diversify their protein sources and more easily achieve nutritional intake recommendations.

Profile and functional properties of seed proteins from six pea (Pisum sativum) genotypes.

Extractability, extractable protein compositions, technological-functional properties of pea (Pisum sativum) proteins from six genotypes grown in Serbia were investigated. Also, the relationship between these characteristics was presented. Investigated genotypes showed significant differences in storage protein content, composition and extractability. The ratio of vicilin:legumin concentrations, as well as the ratio of vicilin + convicilin: Legumin concentrations were positively correlated with extractability. Our data suggest that the higher level of vicilin and/or a lower level of legumin have a positive influence on protein extractability. The emulsion activity index (EAI) was strongly and positively correlated with the solubility, while no significant correlation was found between emulsion stability (ESI) and solubility, nor between foaming properties and solubility. No association was evident between ESI and EAI. A moderate positive correlation between emulsion stability and foam capacity was observed. Proteins from the investigated genotypes expressed significantly different emulsifying properties and foam capacity at different pH values, whereas low foam stability was detected. It appears that genotype has considerable influence on content, composition and technological-functional properties of pea bean proteins. This fact can be very useful for food scientists in efforts to improve the quality of peas and pea protein products.

Utilization of plant-based protein-polyphenol complexes to form and stabilize emulsions: Pea proteins and grape seed proanthocyanidins.

Plant-based proteins and polyphenols are increasingly being explored as functional food ingredients. Colloidal complexes were prepared from pea protein (PP) and grape seed proanthocyanidin (GSP) and the ability of the PP/GSP complexes to form and stabilize oil-in-water emulsions were investigated. The main interactions between PP and GSP were hydrogen bonding. The stability of PP-GSP complexes to environmental changes were studied: pH (2-9); ion strength (0-0.3 M); and temperature (30-90 °C). Emulsions produced using PP-GSP complexes as emulsifiers had small mean droplet diameters (~200 nm) and strongly negative surface potentials (~-60 mV). Compared to PP alone, PP-GSP complexes slightly decreased the isoelectric point, thermostability, and salt stability of the emulsions, but increased their storage stability. The presence of GSP gave the emulsions a strong salmon (red-yellow) color, which may be beneficial for some specific applications. These results may assist in the creation of more efficacious food-based strategies for delivering proanthocyanidins.

Formation and characterization of protein-based films from yellow pea (Pisum sativum) protein isolate and concentrate for edible applications.

This study investigated the properties of films or bioplastics fabricated using a wet processing method from yellow pea protein isolate (YPI) and protein concentrate (YPC) for potential application in food packaging. The wet processing method included mixing the protein with water and glycerol followed by casting and drying the films in a humidity- and temperature-controlled chamber. Whey protein isolate (WPI) and a film from a blend of equal amounts of YPI and WPI, labelled as YPI + WPI, were also studied. Fourier transform-infra red analysis revealed that films from YPI, YPC, WPI and YPI + WPI were formed by protein polymerisation with the plasticiser, glycerol, via hydrophobic and hydrophilic interactions. The protein films had contact angles of <90° demonstrating that they had a hydrophilic surface, with YPC < YPI < YPI + WPI < WPI. The pattern of ultraviolent light transmission of the films was WPI > YPC > YPI + WPI > YPI, whereas the mechanical and thermal resilience of films formulated from YPI, YPC and the protein blend were comparable to the properties of WPI-based films. The findings demonstrate that yellow pea proteins can be used as biomaterials to develop protein and protein-blend films or bioplastics for food packaging and edible applications.

The ability to store energy in pea protein gels is set by network dimensions smaller than 50nm.

The objective of this study was to identify which length scales set the ability to elastically store energy in pea protein network structures. Various network structures were obtained from pea proteins by varying the pH and salt conditions during gel formation. The coarseness of the network structure was visualized by the use of confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) and ranked from least coarse to most coarse networks. Least coarse networks were formed at a pH away from the isoelectric point (IEP) of pea proteins, and at a low ionic strength, whereas more coarse networks were formed at pH values close to the IEP and at a high ionic strength during gel formation. Mechanical deformation properties of the gels such as elastically stored (recoverable) energy, Young's moduli (stiffness of gels), fracture stress (gel strength), and fracture strain (brittleness of the gels) were measured by the use of a texture analyzer and correlated to the coarseness of the networks structure. The influence of coarseness on the ability of the networks to elastically store energy was observed for length scales below 50nm. The findings show that elastically stored energy of pea protein gels can be modulated via the creation of different network structures below 50nm length scales. The results from this study contribute to a better understanding of the dimensions that set the ability to elastically storage in pea protein gels. If the ability of pea proteins to store energy can be understood, products can be better tailored for consumers.

A time-resolved investigation at multiple-length scales of the structure of liquid foam stabilized by albumins from pea.

HYPOTHESIS: The structural details of foams made with pea albumins are affected by the pH of the initial solution and followed heat treatment. EXPERIMENTS: An in situ, time-resolved investigation of foams prepared with pea albumins was conducted using small-angle neutron scattering (SANS) in combination with imaging and conductance measurements. Solutions were tested at pH three pH values (3, 4.5, and 8) before and after heating (90 °C for 1 and 5 min). FINDINGS: The characteristic structures present in the foam from the nano to the meso-scale differed during drainage depending on solution pH. Foams obtained at pH 3, had the largest bubble radius and thinnest plateau border, as well as the highest extent of liquid drainage. At pH 4.5, close to the isoelectric point of the proteins, foams displayed similar bubbles' behavior to those at pH 8, but with the largest film thickness. In this case, the proteins were extensively aggregated. Heating of the solutions prior to foaming did not significantly affect the foam aging regardless of pH. The quantification of specific surface areas and film thickness over time without sample disruption shows to be a powerful approach to designing foam structures.