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.

Analysis of pea mutants reveals the conserved role of FRUITFULL controlling the end of flowering and its potential to boost yield.

Monocarpic plants have a single reproductive phase in their life. Therefore, flower and fruit production are restricted to the length of this period. This reproductive strategy involves the regulation of flowering cessation by a coordinated arrest of the growth of the inflorescence meristems, optimizing resource allocation to ensure seed filling. Flowering cessation appears to be a regulated phenomenon in all monocarpic plants. Early studies in several species identified seed production as a major factor triggering inflorescence proliferative arrest. Recently, genetic factors controlling inflorescence arrest, in parallel to the putative signals elicited by seed production, have started to be uncovered in Arabidopsis, with the MADS-box gene FRUITFULL (FUL) playing a central role in the process. However, whether the genetic network regulating arrest is also at play in other species is completely unknown. Here, we show that this role of FUL is not restricted to Arabidopsis but is conserved in another monocarpic species with a different inflorescence structure, field pea, strongly suggesting that the network controlling the end of flowering is common to other plants. Moreover, field trials with lines carrying mutations in pea FUL genes show that they could be used to boost crop yield.

In vitro protein digestibility of RuBisCO-enriched wheat dough: a comparative study with pea and gluten proteins.

Growing demand for sustainable, plant-based protein sources has stimulated interest in new ingredients for food enrichment. This study investigates the nutritional and digestive implications of enriching wheat dough with RuBisCO, in comparison to pea protein-enriched and gluten-enriched doughs. The protein quality and digestibility of these enriched doughs were analysed through dough characterization, in vitro digestion experiments and biochemical analysis of digesta. Our findings indicate that an enrichment at 10% of RuBisCO or pea proteins improves the chemical score and the in vitro PDCAAS (IV-PDCAAS) score of wheat dough as compared to the control dough. Digestibility assays suggest that RuBisCO introduction modifies the protein hydrolysis kinetics: the nitrogen release is lower during gastric digestion but larger during intestinal digestion than other samples. The analysis of the protein composition of the soluble and insoluble parts of digesta, using size-exclusion chromatography, reveals that the protein network in RuBisCO-enriched dough is more resistant to gastric hydrolysis than the ones of other doughs. Indeed, non-covalently bound peptides and disulfide-bound protein aggregates partly composed of RuBisCO subunits remain insoluble at the end of the gastric phase. The digestion of these protein structures is then mostly performed during the intestinal phase. These results are also discussed in relation to the digestive enzymatic cleavage sites, the presence of potential enzyme inhibitors, the protein aggregation state and the secondary structures of the protein network in each dough type.

Enhancing the functionality of pea proteins by conjugation with propylene glycol alginate via transacylation reaction assisted with ultrasonication.

Pea proteins lack the desirable functional characteristics for food and beverage applications. In this study, transacylation reaction assisted with ultrasonication was used to glycate pea proteins with propylene glycol alginate to enhance their functional properties. The reaction was carried out at pH 11.0 for different pea protein isolate: propylene glycol alginate mass ratios and time durations in a sonic bath at 40 °C. Glycation was confirmed in gel electrophoresis, and ultrasonication enhanced the glycation, with optimal degrees of glycation observed at 45 min reaction time and mass ratios of 2:1 (37.73%) and 1:1 (35.96%). The transacylation reaction increased random coil content of pea proteins by 28% and enhanced their solubility by 2.02 times at pH 7.0, water holding capacity by >50% at pH 7.0, foaming properties, emulsifying properties, and heat stability. This study offers a novel approach that can enhance functionality and applicability of pea proteins.

Acid-induced pea protein gels pretreated with media milling: Gelling properties and the formation mechanism.

This study explored the effect of stirred media mill (SMM) processing on the acid-induced gelling properties of pea protein. Results showed that SMM treatment enhanced the gel strength from 75.06 g to 183.89 g and increased the water holding capacity from 46.64 % to 73.50 %. The minimum gelation concentration achieved for SMM-treated pea protein was 4 %, significantly lower than that of heat-pretreated pea protein (9 %). SMM decreased protein aggregate size from 104 µm to 180 nm. Microscopy analysis revealed that the small aggregates facilitated the formation of uniform gel networks with tight connections. Linear rheology indicated that small protein aggregates resulted in slower gelation rates with a higher G' for the formed gels. The SMM-pretreated protein gel showed strain hardening, shear thinning behaviors, and satisfactory stability to withstand large-amplitude oscillatory shear. Overall, SMM emerges as a promising technology for producing protein gel products with strong mechanical attributes and customizable rheological properties.

In vitro gastrointestinal simulated digestion of three plant proteins: determination of digestion rate, free amino acids and peptide contents.

Cassava protein (CP), barley protein (BP) and yellow pea protein (YPP) are important nutrient and integral constituent of staple in pet foods. It is known that the digestion of proteins directly influences their absorption and utilisation. In the present work, we performed in vitro simulated gastrointestinal digestion of three plant proteins as a staple for dog and cat food. The digestion rate of CP, BP and YPP in dog food was 56.33 ± 0.90%, 48.53 ± 0.91%, and 66.96 ± 0.37%, respectively, whereas the digestion rate of CP, BP, and YPP in cat food was 66.25 ± 0.72%, 43.42 ± 0.83%, and 58.05 ± 0.85%, respectively. Using SDS-polyacrylamide gel electrophoresis to determine the molecular weight (MW) of each protein and the products of their digestion, it was revealed that MW of digestion samples decreased, and MW during the small intestine phase was lower than that during the gastric phase. Peptide sequences of digested products were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and it was found that the total number of peptides in the small intestine digestion samples was higher than that in the gastric phase samples. The MW of peptides obtained from CP was within the range of 1000-1500 Da, while MW of peptides derived from BP and YPP was within the range of 400-2000 Da. In addition, free amino acids were mainly produced in the small intestine phase. Furthermore, the percentage of essential amino acids in the small intestine phase (63 ~ 82%) was higher than that in the gastric phase (37 ~ 63%). Taken together, these findings contribute to the current understanding of the utilisation of plant proteins in dog and cat foods and provide important insights into the selection and application of plant proteins as a staple in dog and cat foods.

Pea protein [Pisum sativum] as stabilizer for oil/water emulsions.

A map of stability for various water/oil/pea protein compositions has been plotted from the numerous reported results. Two clear regions of stability were identified. High internal oil phase emulsions with 70-80%, v/v oil content stabilized by total pea protein concentration <2.5%, w/v showed stability. Low oil content of 10-30%, v/v for a range of total pea protein concentrations >0.5%, w/v have also been identified as stable. Intermediate oil content and pea protein concentrations >4% w/v are unexplored regions and are likely to be areas of fruitful future research. The wide range of stability suggests that different stabilization mechanisms could be important for different compositions and careful consideration has to be taken to avoid oversimplification. Both stabilization with particles, i.e. Pickering emulsions, and protein unfolding have been suggested as mechanisms. The diverse way of describing stability makes it difficult to intercompare results in different studies. A summary of different oil types used have been presented and several properties such as dynamic viscosity, density, the dielectric constant and interfacial tension have been summarized for common vegetable oils. The type of vegetable oil and emulsion preparation techniques were seen to have rather little effect on emulsion stability. However, the different extraction methods and processing of the pea material had more effect, which could be attributed to changing composition of different proteins and to the states of aggregation and denaturing. Careful consideration has to be taken in the choice of extraction method and an increased understanding of what contributes to the stability is desirable for further progress in research and eventual product formulation.

Modification of soy protein isolate and pea protein isolate by high voltage dielectric barrier discharge (DBD) atmospheric cold plasma: Comparative study on structural, rheological and techno-functional characteristics.

The modification in structural, rheological, and techno-functional characteristics of soy and pea protein isolates (SPI and PPI) due to dielectric barrier discharge cold plasma (DBD-CP) were assessed. The increased carbonyl groups in both samples with cold plasma (CP) treatment led to a reduction in free sulfhydryl groups. Moreover, protein solubility of treated proteins exhibited significant improvements, reaching up to 59.07 % and 41.4 % for SPI and PPI, respectively, at 30 kV for 8 min. Rheological analyses indicated that storage modulus (G') was greater than loss modulus (G″) for CP-treated protein gels. Furthermore, in vitro protein digestibility of SPI exhibited a remarkable improvement (4.78 %) at 30 kV for 6 min compared to PPI (3.23 %). Spectroscopic analyses, including circular dichroism and Fourier Transform-Raman, indicated partial breakdown and loss of α-helix structure in both samples, leading to the aggregation of proteins. Thus, DBD-CP induces reactive oxygen species-mediated oxidation, modifying the secondary and tertiary structures of samples.

Insight into the improvement mechanism of gel properties of pea protein isolate based on the synergistic effect of cellulose nanocrystals and calcium ions.

Here, the influence and potential mechanism by which cellulose nanocrystals (CNC) collaborated with Ca2+ enhancing the heat-induced gelation of pea protein isolate (PPI) were investigated. It was found that the combination of 0.45% CNC and 15 mM Ca2+ synergistically increased the gel strength (from 14.18 to 65.42 g) and viscoelasticity of PPI while decreased the water holding capacity. The improved particle size, turbidity, and thermostability as well as the reduced solubility, crystallinity, and gel porosity were observed in CNC/CaCl2 composite system. CNC fragments bind to specific amino acids in 11S legumin and 7S vicilin mainly through hydrogen bonding and van der Waals forces. Moreover, changes in the protein secondary structure and enhancement of the molecular interaction induced by CNC and Ca2+ could favor the robust gel network. The results will provide a new perspective on the functional regulation of pea protein and the creation of pea protein gel-based food.

Ultra-stable pickering emulsion stabilized by anisotropic pea protein isolate-fucoidan conjugate particles through Maillard reaction.

Bio-based emulsifiers hold significant importance in various industries, particularly in food, cosmetics, pharmaceuticals and other related fields. In this study, pea protein isolate (PPI) and fucoidan (FUD) were conjugated via the Maillard reaction, which is considered safe and widely used in the preparation of food particle. The PPI-FUD conjugated particles exhibit an anisotropic non-spherical structure, thereby possessing a high detachment energy capable of preventing emulsion coalescence and Ostwald ripening. Compared to emulsions previously prepared in other studies (< 500 mM), the Pickering emulsion stabilized by PPI-FUD conjugate particles demonstrates outstanding ionic strength resistance (up to 5000 mM). Furthermore, when encapsulating curcumin, the Pickering emulsion protects the curcumin from oxidation. Additionally, the formulated emulsions demonstrated the capability to incorporate up to 60 % (v/v) oil phase, revealing remarkable performance in terms of storage stability, pH stability, and thermal stability.

The effects of resonance acoustic mixing modulation on the structural and emulsifying properties of pea protein isolate.

The effects of resonant acoustic mixing (RAM) with different treatment times (0, 5, 10, 15, 20 and 30 min) on the structural and emulsifying properties of pea protein isolate (PPI) were investigated for the first time. Increasing the RAM treatment time from 0 to 20 min decreased the α-helix/ß-sheet ratio and particle size of the PPI samples by 37.84 % and 46.44 %, respectively, accompanied by an increase in solubility from 54.79 % to 71.80 % (P < 0.05). Consequently, the emulsifying activity index of PPI (from 10.45 m2/g to 14.2 m2/g) and the physical stability of RAM-PPI emulsions were effectively enhanced, which was confirmed by the small and uniformly distributed oil droplets in the micrographs of the emulsions. However, excessive RAM treatment (30 min) diminished the effectiveness of the aforementioned improvements. Therefore, obviously enhanced solubility and emulsifying properties of PPI can be attained through proper RAM treatment (15-20 min).

Konjac glucomannan-assisted fabrication of stable emulsion-based oleogels constructed with pea protein isolate and its application in surimi gels.

Konjac glucomannan (KGM) is widely used as a stabilizer for the structuring of highly unsaturated oils. This study aimed to investigate the changes in structure and functional properties of soybean oil - based oleogels (emulsion template method) prepared with different amounts of KGM-modified pea isolate protein (PPI). The findings revealed that the oleogels formed three - dimensional networks through van der Waals interactions and hydrogen bonding between the stretched PPI and KGM. As the amount of KGM increased, the oil droplets were more uniformly dispersed within the continuous PPI - KGM rigid network, especially when the ratio of PPI to KGM was 4:1. This formulation also showed the highest thixotropy (73.2 %) and the best oil binding capacity (94 %). Cryo - SEM revealed that the oleogel - prepared surimi gels successfully enclosed oil droplets in a dense matrix through a dual stabilization mechanism. Additionally, the incorporation of oleogels significantly improved the textural properties of surimi in comparison to directly adding oil.

Fermentation of Texturized Pea Protein in Combination with Proteases for Aroma Development in Meat Analogues.

The potential use of texturized pea protein in meat analogues was investigated by comparing the effects of fermentation on pea and myofibrillar pork proteins in a model system including additives, microbial starters, and proteases. Model fermentation was controlled for 15 days by a pH decrease and microbial count and free amino acid increase. Besides, volatile production and sensory properties were evaluated at the end of fermentation. Protein type affected free amino acid generation and volatile profile. Models supplemented with proteases showed an increase in amino-acid-derived compounds (branched aldehydes and alcohols) and fruity odor notes. During fermentation, protease addition significantly reduced the production of linear aldehydes (pentanal, hexanal, and octanal) in vegetal models, while pyrazine compounds were not affected. This changes in the volatile profile reduced the legume beany odor but increased the perception of toasted cereal-like notes generated by the texturization process.

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.

Improving the Three-Dimensional Printability of Potato Starch Loaded onto Food Ink.

This study focuses on improving the 3D printability of pea protein with the help of food inks designed for jet-type 3D printers. Initially, the food ink base was formulated using nanocellulose-alginate with a gradient of native potato starch and its 3D printability was evaluated. The 3D-printed structures using only candidates for the food ink base formulated with or without potato starch exhibited dimensional accuracy exceeding 95% on both the X and Y axes. However, the accuracy of stacking on the Z-axis was significantly affected by the ink composition. Food ink with 1% potato starch closely matched the CAD design, with an accuracy of approximately 99% on the Z-axis. Potato starch enhanced the stacking of 3D-printed structures by improving the electrostatic repulsion, viscoelasticity, and thixotropic behavior of the food ink base. The 3D printability of pea protein was evaluated using the selected food ink base, showing a 46% improvement in dimensional accuracy on the Z-axis compared to the control group printed with a food ink base lacking potato starch. These findings suggest that starch can serve as an additive support for high-resolution 3D jet-type printing of food ink material.

Constructing a strongly interacting Pea-Cod binary protein system by introducing metal cations toward enhanced gelling properties.

Developing prospective plant-animal binary protein systems with desirable nutritional and rheological properties stands as a significant and challenging pursuit within the food industry. Our understanding of the effect of adding salt on the aggregation behavior of food proteins is currently based on single model protein systems, however, this knowledge is rather limited following binary protein systems. Herein, various ionic strength settings are used to mitigate the repulsive forces between pea-cod mixed proteins during the thermal process, which further benefits the construction of a strengthened gel network. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) collectively demonstrated that larger heat-induced protein aggregates were formed, which increased in size with higher ionic strength. In the presence of 2.5 mM CaCl2 and 50 mM NaCl, the disulfide bonds significantly increased from 19.3 to 27.53 and 30.5 µM/g, respectively. Notably, similar aggregation behavior could be found when introducing 2.5 mM CaCl2 or 25 mM NaCl, due to the enhanced aggregation tendency by specific binding of Ca2+ to proteins. With relevance to the strengthened cross-links between protein molecules, salt endowed composite gels with preferable gelling properties, evidenced by increased storage modulus. Additionally, the gelling temperature of mixed proteins decreased below 50 °C at elevated ionic strength. Simultaneously, the proportion of network proteins in composite gels increased remarkably from 82.05 % to 93.61 % and 92.31 % upon adding 5.0 mM CaCl2 and 100 mM NaCl, respectively. The findings provide a valuable foundation for designing economically viable and health-oriented plant-animal binary protein systems.

Chemical acylation of pea protein isolate hydrolysate with fatty acid N-hydroxysuccinimide esters: Effect on structure and functional properties.

Applications of pea protein in the food industry have been greatly restricted by its poor functional properties. In order to solve this problem, a novel technique combining enzymatic hydrolysis and fatty acid acylation has been applied in this work to construct a pea protein-fatty acid covalent complex that aims to improve its functional properties. The processed pea protein with increased water solubility tends to decrease the chance of self-aggregation. Additionally, emulsifying and antioxidant properties have also been found after this process. On top of that, the modified pea protein has been characterized by Fourier transform infrared and circular dichroism spectroscopy. These results demonstrate that these properties were mainly caused by the acylation of the amino group from hydrolyzed pea protein and the carboxyl group from the fatty acid. The enzymatic hydrolysis/fatty acid acylation research provides insights into manufacturing high-quality functional lipoproteins from inexpensive pea protein for the food industry.

Efficacy of an oral suspension containing xyloglucan and pea proteins on a murine model of gastroesophageal reflux disease.

Gastroesophageal reflux disease (GERD) is the most common foregut disease, affecting about 20% of the adult population. Esophageal epithelial barrier plays a fundamental role in the pathophysiology of GERD; however, pharmacological therapies mainly aim to reduce the acidity of the gastroesophageal environment rather than to protect esophageal tissue integrity. This study aims to evaluate the efficacy of an oral solution containing xyloglucan and pea proteins (XP) in reestablishing gastroesophageal tissue integrity and biochemical markers. To induce GERD, C57BL/6 mice were alternatively overfed and fasted for 56 days and then treated with XP, sodium alginate, omeprazole, or omeprazole+XP twice daily for 7 days. Gastric pain and inflammatory markers were evaluated after 3 and 7 days of treatment. After sacrifice, the esophagi and stomachs were surgically removed for macroscopic and histological examination. Gastric pain was significantly reduced at days 3 and 7 by XP, omeprazole, and omeprazole+XP, while alginates were ineffective at day 3. XP was able to diminish gastric macroscopic damage and demonstrated the same efficacy as omeprazole in reducing esophageal damage. XP significantly reduced histological damage, with an efficacy comparable to that of omeprazole, but superior to alginates. Inflammatory markers were significantly reduced by XP, with superior efficacy compared with alginates at day 7. Interestingly, XP was also able to significantly increase gastric pH. This study demonstrated that XP restored gastric homeostasis, improved esophageal integrity, and decreased inflammation and pain with a similar efficacy to omeprazole and greater than alginates.

Ingestion of a variety of non-animal-derived dietary protein sources results in diverse postprandial plasma amino acid responses which differ between young and older adults.

Whole-body tissue protein turnover is regulated, in part, by the postprandial rise in plasma amino acid concentrations, although minimal data exist on the amino acid response following non-animal-derived protein consumption. We hypothesised that the ingestion of novel plant- and algae-derived dietary protein sources would elicit divergent plasma amino acid responses when compared with vegan- and animal-derived control proteins. Twelve healthy young (male (m)/female (f): 6/6; age: 22 ± 1 years) and 10 healthy older (m/f: 5/5; age: 69 ± 2 years) adults participated in a randomised, double-blind, cross-over trial. During each visit, volunteers consumed 30 g of protein from milk, mycoprotein, pea, lupin, spirulina or chlorella. Repeated arterialised venous blood samples were collected at baseline and over a 5-h postprandial period to assess circulating amino acid, glucose and insulin concentrations. Protein ingestion increased plasma total and essential amino acid concentrations (P < 0·001), to differing degrees between sources (P < 0·001), and the increase was further modulated by age (P < 0·001). Postprandial maximal plasma total and essential amino acid concentrations were highest for pea (2828 ± 106 and 1480 ± 51 µmol·l-1) and spirulina (2809 ± 99 and 1455 ± 49 µmol·l-1) and lowest for chlorella (2053 ± 83 and 983 ± 35 µmol·l-1) (P < 0·001), but were not affected by age (P > 0·05). Postprandial total and essential amino acid availabilities were highest for pea, spirulina and mycoprotein and lowest for chlorella (all P < 0·05), but no effect of age was observed (P > 0·05). The ingestion of a variety of novel non-animal-derived dietary protein sources elicits divergent plasma amino acid responses, which are further modulated by age.

Structure-property relationship of pea protein fibrils in stabilization of HIPEs and the encapsulation, protection, controlled release and oral delivery of carotenoids for alleviating intestinal inflammation.

Increasing attentions are paid to high internal phase emulsions (HIPEs) due to their unique properties. In this study, pea protein-based fibrils were used as emulsifier to stabilize HIPEs. We demonstrated that the molecular assembly pathway and interfacial behavior of pea protein-based fibrils are affected by ionic strength. And the increased abundance of highly flexible worm-like nanofibrils facilitated their adsorption and packing on oil droplets, resulting in improved emulsion properties to stabilize the HIPEs with the internal phase volume fraction as high as 90%. Based on this, high loading content of carotenoids up to 0.05 wt% in the prepared HIPEs, protection of their stability against heating, UV and iron ions, and significantly increased bio-accessibilities of the carotenoids were realized. Animal studies using a mouse model of DSS-induced colitis revealed that carotenoid loaded HIPEs can alleviate the colon injury, by downregulating the expression of inflammatory cytokines, and promoting intestinal barrier function. This work will deepen the understanding of the formation of pea protein fibrils and provide a reference for the rational use of carotenoid loaded HIPEs in IBD management.