Effect of Heat Treatment on Yellow Field Pea (Pisum sativum) Protein Concentrate Coupled with Membrane Ultrafiltration on Emulsification Properties of the Isolated >50 kDa Proteins.

The aim of this paper was to determine the emulsification properties of protein aggregates obtained from heat pretreated yellow field pea protein concentrate (PPC). PPC dispersions were prepared in distilled water (adjusted to pH 3.0, 5.0, 7.0, or 9.0), heated in a water bath (100 °C) for 30 min, centrifuged and the supernatant passed first through a 30 kDa membrane and, then, the first retentate (>30 kDa) through a 50 kDa membrane. The 50 kDa membrane separation yielded a second retentate (>50 kDa proteins), which was isolated for emulsification studies. The near UV circular dichroic spectra of the protein samples showed more unfolded structures at pH 3.0 and 5.0 than at pH 7.0 and 9.0. The presence of small and spherical oil droplets of emulsions stabilized by the >50 kDa proteins at pH 3.0, 7.0, and 9.0 was confirmed by confocal laser scanning microscopy images. Emulsions stabilized at pH 7.0 and 9.0 had a narrower size distribution range than at pH 3.0 and 5.0. A narrow oil droplet size distribution range and lower interfacial protein concentrations of the emulsions stabilized by the >50 kDa proteins were observed at the corresponding pH of the heat treatment when compared to other pH values. Emulsions stabilized by the >50 kDa proteins exhibited a relatively low flocculation and coalescence index, which infers relative stability. The results from this work suggest that heat pretreatment of the PPC led to the formation of new protein aggregates, especially FT9 with enhanced emulsification properties, at some of the test conditions when compared to the unheated PPC.

Quantitative Structure-Activity Relationship Modeling of Pea Protein-Derived Acetylcholinesterase and Butyrylcholinesterase Inhibitory Peptides.

The aim of this work was to determine the structural requirements for peptides that inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities. The data set used consisted of 19 oligopeptides that had been identified through mass spectrometry analysis of enzymatic digests of yellow field pea protein. The structure-function relationship was analyzed by partial least squares regression using the 5z scores. A nine-component model was created from 16 peptides for AChE inhibitory peptides (Q2 = 67.2% and R2 = 0.9974), while three data sets were prepared for BuChE inhibitory peptides to improve the quality of the models (Q2 = 26.7-46.4% and R2 = 0.9577-0.9958). The most active peptides from the PLS models have threonine, leucine, alanine, and valine at the N terminal, asparagine, histidine, proline, and arginine at the second position, with aspartic acid and serine at the third, and arginine at the C terminal.

Yellow Field Pea Protein (Pisum sativum L.): Extraction Technologies, Functionalities, and Applications.

Yellow field peas (Pisum sativum L.) hold significant value for producers, researchers, and ingredient manufacturers due to their wealthy composition of protein, starch, and micronutrients. The protein quality in peas is influenced by both intrinsic factors like amino acid composition and spatial conformations and extrinsic factors including growth and processing conditions. The existing literature substantiates that the structural modulation and optimization of functional, organoleptic, and nutritional attributes of pea proteins can be obtained through a combination of chemical, physical, and enzymatic approaches, resulting in superior protein ingredients. This review underscores recent methodologies in pea protein extraction aimed at enhancing yield and functionality for diverse food systems and also delineates existing research gaps related to mitigating off-flavor issues in pea proteins. A comprehensive examination of conventional dry and wet methods is provided, in conjunction with environmentally friendly approaches like ultrafiltration and enzyme-assisted techniques. Additionally, the innovative application of hydrodynamic cavitation technology in protein extraction is explored, focusing on its prospective role in flavor amelioration. This overview offers a nuanced understanding of the advancements in pea protein extraction methods, catering to the interests of varied stakeholders in the field.

Physicochemical and Functional Properties of Membrane-Fractionated Heat-Induced Pea Protein Aggregates.

This study was carried out to investigate the effect of heat pre-treatment of pea proteins at different pH values on the formation of functional protein aggregates. A 10% (w/v) aqueous mixture of pea protein concentrate (PPC) was adjusted to pH 3.0, 5.0, 7.0, or 9.0 followed by heating at 100°C for 30 min, cooled and centrifuged. The supernatant was sequentially passed through 30 and 50 kDa molecular weight cut-off membranes to collect the <30, 30-50, and >50 kDa fractions. The >50 kDa fractions from pH 3.0 (FT3), 5.0 (FT5), 7.0 (FT7), and 9.0 (FT9) treatments had >60% protein content in contrast to the ≤20% for the <30 and 30-50 kDa fractions. Therefore, the >50 kDa fractions were collected and then compared to the untreated PPC for some physicochemical and functional properties. Protein aggregation was confirmed as the denaturation temperature for FT3 (124.30°C), FT5 (190.66oC), FT7 (206.33oC) and FT9 (203.17oC) was significantly (p < 0.05) greater than that of PPC (74.45oC). Scanning electron microscopy showed that FT5 had a compact structure like PPC while FT3, FT7, and FT9 contained a more continuous network. In comparison to PPC, the >50 kDa fractions showed improved solubility (>60%), oil holding capacity (~100%), protein content (~7%), foam capacity (>10%), foam stability (>7%), water holding capacity (>16%) and surface hydrophobicity (~50%). Least gelation concentration of PPC (18%), FT3 (25%), FT5 (22%), FT7 (22%), and FT9 (25%) was improved to 16, 18, 20, 16, and 18%, respectively, after addition of NaCl.

Acetylcholinesterase and butyrylcholinesterase inhibitory activities of antioxidant peptides obtained from enzymatic pea protein hydrolysates and their ultrafiltration peptide fractions.

This study optimized the enzymatic hydrolysis of yellow field pea proteins using alcalase (ACH), chymotrypsin (CHH), flavourzyme (FZH), pancreatin (PCH), pepsin (PEH), and trypsin (TPH) to obtain hydrolysates and ultrafiltered fractions (<1, 1-3, 3-5 and 5-10 kDa) that possess antioxidant plus acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities. The hydrolysates exhibited varying degrees of radical scavenging and inhibition of linoleic acid peroxidation, as well as cholinesterase inhibition activities but the potency generally improved by >10% after UF separation into peptide fractions. ACH, FZH, and PEH exhibited significantly (p < .05) higher (20%-30% increases) radical scavenging activities than the other hydrolysates. The 1 and 3 kDa UF fractions of ACH, FZH, and PEH inhibited ~20%-30% AChE activity, while ACH, PCH, TPH, and PEH inhibited ~20%-40% BuChE activity. We conclude that the pea protein hydrolysates and their peptide fractions possess multifunctional properties with potential use against neurodegenerative disorders. PRACTICAL APPLICATIONS: Alzheimer's disease (AD) has multiple pathological pathways in addition to the loss of acetylcholine (ACh) catalyzed by acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The presence of severe oxidative stress triggered by lipid peroxidation and formation of free radicals is a common trait in AD patients. The concept of AChE and BuChE inhibition as an approach toward AD amelioration involves the use of compounds with a similar structure to ACh, the natural substrate. Peptides derived from food proteins consist of ester bonds with structural similarity to ACh and theoretically possess the ability to interact with AChE and BuChE. Results from the present study imply that pea protein-derived peptides are potential candidates for use as inhibitors of AChE and BuChE activities, with application in the prevention and management of AD.

In vitro inhibition of acetylcholinesterase activity by yellow field pea (Pisum sativum) protein-derived peptides as revealed by kinetics and molecular docking.

Compounds with structural similarities to the neurotransmitter (acetylcholine) are mostly used to inhibit the activity of acetylcholinesterase (AChE) in Alzheimer's disease (AD) therapy. However, the existing drugs only alleviate symptoms of moderate to mild conditions and come with side effects; hence, the search is still on for potent and safer options. In this study, High performance liquid chromatography (HPLC) fractionations of AChE-inhibitory pea protein hydrolysates obtained from alcalase, flavourzyme and pepsin digestions were carried out followed by sequence identification of the most active fractions using mass spectrometry. Subsequently, 20 novel peptide sequences identified from the active fractions were synthesized and five peptides, QSQS, LQHNA, SQSRS, ETRSQ, PQDER (IC50 = 1.53 - 1.61 µg/mL) were selected and analyzed for ability to change AChE protein conformation (fluorescence emission and circular dichroism), kinetics of enzyme inhibition, and enzyme-ligand binding configurations using molecular docking. The kinetics studies revealed different inhibition modes by the peptides with relatively low (<0.02 mM and <0.1 mM) inhibition constant and Michaelis constant, respectively, while maximum velocity was reduced. Conformational changes were confirmed by losses in fluorescence intensity and reduced α-helix content of AChE after interactions with different peptides. Molecular docking revealed binding of the peptides to both the catalytic anionic site and the peripheral anionic site. The five analyzed peptides all contained glutamine (Q) but sequences with Q in the penultimate N-terminal position (LQHNA, SQSRS, and PQDER) had stronger binding affinity. Results from the different analysis in this study confirm that the peptides obtained from enzymatic digestion of pea protein possess the potential to be used as novel AChE-inhibitory agents in AD management.