Pea Protein Extracts Research Articles

EFEKTIVITAS BUNGA TELANG (CLITORIA TERNATEA) TERHADAP RESPON GLIKEMIK: SISTEMATIK REVIEW

Background: Diabetes mellitus is a chronic disease that is globally prevalent and has reached a very high prevalence rate. In 2021, more than 422 million people worldwide will suffer from diabetes. Various databases have reported that one of the ornamental plants, namely the butterfly pea flower (Clitoria Ternatea), has quite a hypoglycemic effect. 
 Purpose: The aim of this systematic research is to help in understanding the potential of butterfly pea flowers as an antidiabetic agent.
 Methods: The research method we used in this systematic review search was the PRISMA method. This article searches literature sources in Pubmed and Google Scholar databases from 2013 to 2023 using appropriate inclusion criteria. Clitoria Ternatea has excellent anti-diabetic properties.
 Result: In vitro analysis showed that water extract of butterfly pea flower (Clitoria Ternatea), ethanol extract, and protein extract effectively inhibited the α-amylase enzyme, hemoglobin glycosylation, and AGE formation. They also show increased glucose absorption.
 Conclusion: This explains that CTE can suppress postprandial glucose and insulin responses induced by sucrose through inhibiting intestinal sucrose. It can be concluded that butterfly pea flowers (Clitoria Ternatea) are effective against glycemic responses in the body.

Pea protein extraction method impacts the protein (micro)structural organisation and in vitro digestion kinetics.

There is increasing interest in including pulse proteins into food products due to their nutrient-rich and sustainable character. However, little is known regarding the consequences of different extraction approaches on the pulse protein structure and the subsequent protein (micro)structural organization and protein digestion kinetics. Therefore, three green pea protein extracts were created: (i) cooking followed by cotyledon cell isolation, (ii) alkaline extraction followed by isoelectric precipitation, or (iii) salt extraction, and compared to the original pea flour as well as to sodium caseinate. The results showed that encapsulated, denatured protein inside pea cotyledon cells presented the (s)lowest digestion, while accessible and more native protein (e.g., pea flour, pea protein salt extract) presented much faster and higher digestion. Moreover, the alkali extracted pea protein was denatured to some extent, significantly lowering in vitro digestion kinetics. In the second part, three different in vitro approaches were applied to digest the salt extracted pea protein. Semi-dynamic gastric digestion approaches simulate in vivo conditions more closely which especially impacted the rate of digestion.

Evaluation of the Efficacy of Xyloglucan, Pea Protein and Opuntia ficus-indica Extract in a Preclinical Model of Psoriasis

Psoriasis is a chronic inflammatory skin disease characterized by epidermal gene abnormalities, epidermal barrier defects and inflammation. Corticosteroids are considered to be standard treatments, but often come with side effects and lose efficacy with long-term use. Alternative treatments targeting the epidermal barrier defect are needed to manage the disease. Film-forming substances such as xyloglucan, pea protein and Opuntia ficus-indica extract (XPO) have generated interest for their ability to restore skin barrier integrity and may pose an alternative approach to disease management. Thus, the aim of this two-part study was to evaluate the barrier-protective properties of a topical cream containing XPO on the membrane permeability of keratinocytes exposed to inflammatory conditions and compare its efficacy to dexamethasone (DXM) in an in vivo model of psoriasis-like dermatitis. XPO treatment significantly reduced S. aureus adhesion, subsequent skin invasion and restored epithelial barrier function in keratinocytes. Furthermore, the treatment restored the integrity of keratinocytes, reducing tissue damage. In mice with psoriasis-like dermatitis, XPO significantly reduced erythema, inflammatory markers and epidermal thickening with a superior efficacy to dexamethasone. Given the promising results, XPO may represent a novel steroid-sparing therapeutic for epidermal-related diseases such as psoriasis, thanks to its ability to preserve skin barrier function and integrity.

The impact of heating and freeze or spray drying on the interface and foam stabilising properties of pea protein extracts: Explained by aggregation and protein composition

The processing of plant protein extracts can affect the protein structure, leading to altered functional properties. In this work, we evaluated the impact of two common processes in pea protein extraction: heating and drying. Non-heated and heated (5 min at 95 °C) samples were compared, which were either freeze- or spray-dried. These processes led to alterations of the proteins, and resulted in changes of their interface and foam-stabilising properties. A mild protein extraction method was used to preserve the native protein structure during aqueous extraction, allowing the extraction of both albumin and globulin proteins. Spray-drying of these fractions led to higher surface hydrophobicity, which resulted in increased surface activity and stiffer interfacial layers at pH 3.8 and 7.0. The heating step induced aggregation of the globulins, while albumins remained soluble. Here, we demonstrated that the albumins had a dominant effect on the interfacial (rheology and ellipsometry) and foaming properties after heating, as the globulin aggregates were too large for effective interface stabilisation. A similar mechanism was also shown at pH 3.8, where the globulins precipitated, as the pH was close to their pI, while albumins remained soluble. Again, the albumins dictated the interfacial properties, leading to highly stable foams after removing the insoluble globulins. We have shown marginal differences in protein functionality after freeze- or spray-drying. More importantly, the changes in soluble protein composition dictate the protein functionality after heating or pH shifts.

Cottage cheese from blends of fresh green peas (Pisum sativum L.) and dairy milk (pEaneer): Preparation, characterization, and sensory evaluation

Traditional tofu from soybeans provides a bunch of health benefits which makes it a staple option in high-protein diets. Concerns on allergenicity/hypercholesterolemia from animal foods and lactose intolerance from dairy based ones have surged the demand for alternative and sustainable plant-based proteins. In this study, a tofu-like coagulated product named pEaneer was prepared using a blend of fresh green peas and dairy milk. Initially, fresh peas were ground with water in 1:2 (w/v) ratio and used for protein extraction by alkali solubilisation followed by isoelectric precipitation. The process parameters such as pH (6-12), time (40–120 min) and temperature (30–90 °C) were optimized for maximum protein yield using full factorial design. The maximum protein yield of 53.99% w/w was obtained at pH 12/60 °C/40min. The pea protein extract (PPE) obtained under optimized conditions was used to obtain pEaneer with and without dairy milk by blending them in various proportions. Efforts to achieve a desirable appearance and texture for pEaneer using D-optimal design indicated 0.3% (w/v) gellan to be effective. The best pEaneer obtained with 80:20 (w/w) of PPE and dairy milk had 58.01% protein and 16.16% fat vis-à-vis 42.07% protein and 48.78% fat in dairy paneer.

Impact of microfluidization on colloidal properties of insoluble pea protein fractions

Microfluidization is a technique commonly used to disrupt and homogenize dispersions such as oil-in-water emulsions or cellular suspensions. In this study, we investigated its ability to alter the physicochemical properties of plant-derived insoluble protein aggregates such as those found in pea protein extracts. Insoluble pea protein dispersions (5% w/w, pH 7) were homogenized at 25–150 MPa for 1–5 cycles. Increasing the homogenization pressure and cycles decreased the particle size (d43) of the unhomogenized insoluble pea proteins from 180 ± 40 μm to 0.2 ± 0.0 μm (at ≥ 125 MPa), leading to more transparent dispersions. Furthermore, the solubility of the insoluble pea proteins increased from 23 ± 1% to 86 ± 4%. Treatments with chaotropic agents, dithiothreitol and urea, revealed that insoluble pea protein aggregates were stabilized not only by disulphide bonds but also by hydrogen bonds and hydrophobic interactions. These molecular interactions were disrupted by microfluidization. The study provides insights into the disruption mechanism of insoluble pea proteins by applying microfluidization and offers a mean to improve their technofunctional properties to facilitate further use in food manufacture.

Pea proteins immunotherapy in peanut allergic mice model

Mice (Balb/c), with peanut allergy induced, were subjected to desensitization therapy with the use of pea protein extract (PE) or isolated globulin fractions: legumin (PL) and vicilin (PV). B- and T-cell responses to peanut proteins were analysed by determination of the IgE, IgG1, and IgG2a antibody levels in plasma and the concentration of IL-4, IFN-gamma and IL-10 cytokines secreted by isolated splenocytes.Conducted studies have demonstrated that immunotherapy with proteins resulted in the decrease of total IgE and peanut-specific IgG1 levels and significantly enhanced synthesis of peanut-specific IgG2a in plasma (ELISA method) and at the cellular level (ELISPOT type B). A successful and effective immunotherapy is related to the shift in profile of lymphocytes from Th2 subpopulation towards Th1 subpopulation. In our studies significant increase in the activity of Th1 lymphocytes was observed in groups desensitized with pea protein extracts (PE) and pea legumin fraction (PL). In these groups, significant statistic decrease in IL-4 secreted and increase in IL-10 level were found.Desensitization method with the use of pea proteins being suggested in the presented studies can be an alternative method for specific immunotherapy for people, especially with strong allergic reaction to peanuts; however, this method needs further studies with mouse model.

Characterisation of odour active compounds along extraction process from pea flour to pea protein extract

Pisum sativum, rich in proteins, represents a main interest for human food. Nevertheless, pea products are underused because of their organoleptic characteristics. The extraction process of the proteins can partly explain the development of the typical flavour.The objective of the present study was to identify odour active compounds and to follow their evolution during four steps of the process (from pea flour to pea protein).Firstly, volatile compounds were extracted by Solvent Assisted Flavour Evaporation from each step and analysed by Gas Chromatography coupled with Mass Spectrometry and Olfactometry. Secondly, the volatile compounds, identified as odour active in pea flour and in pea protein, were followed in the intermediary steps of the process.As hypothesised, it appeared that the flavour profile is evolving during the protein extraction process. The odour active compounds are different between pea flour (first step) and pea protein powder (final step). Some compounds are disappearing earlier or later during the process, whereas some are appearing at different steps. Only few odour active compounds are common to each step.These results lead to a better understanding of the flavour compounds of pea flour and of proteins, and of their evolution during the protein extraction process. Then, these results could permit to adapt, to improve some key parameters or to add some steps at the process in order to better control the beany flavour development.

Vicilin and the basic subunit of legumin are putative chickpea allergens

IgE-mediated reactions to food allergens constitute a major health problem in industrialized countries. Chickpea is consumed in Mediterranean countries, and reportedly associated with IgE-mediated hypersensitivity reactions. However, the nature of allergic reactions to chickpea has not been characterized.A serum pool from paediatric patients allergic to chickpeas was used to detect IgE-binding proteins from chickpea seeds by immunoassay and immunoblot inhibition assay. Protein samples enriched in chickpea legumin and vicilin were obtained by anion exchange chromatography, and were identified by mass spectrometric analysis.IgE-immunoassays of globulin fractions from chickpeas revealed that vicilin (50kDa) and the basic subunit of legumin (20kDa) were bound by IgE from patient sera. Pea and lentil protein extracts strongly inhibited the IgE binding to chickpea globulin.We speculate that vicilin and the basic subunit of legumin are major chickpea allergens. Also, the globulin fraction of chickpea likely cross-reacts with the allergenic proteins of pea and lentil.

Improvement of the Aroma of Pea (Pisum sativum) Protein Extracts by Lactic Acid Fermentation

The formation of a green or beany off-flavor during storage of legume protein extracts limits their application in foods. Pea protein extracts were submitted to lactic acid fermentations to improve the flavor by either reducing off-flavor formation or by masking undesirable green notes. The aroma profiles of untreated pea protein extract (PPE) and fermented pea protein extract (PPEF) were compared to each other and to a commercial whey protein as a benchmark. Kinetic measurements of n-hexanal and n-hexanol formation were used as an indicator for progressing lipid oxidation and storage stability. The nonfermented and fermented pea protein extracts showed a shelf-life comparable to the commercial whey protein reference. Volatiles were identified and quantified using dynamic headspace sampling with subsequent coupled TDS-GC-MS and TDS-GC-olfactometry flavor dilution analysis. A total of 18 odorants with dilution factors equal to or higher than 100 were determined in PPE and 17 in PPEF. Altogether, 23 highly odor-active compounds were identified according to their mass spectra, odor impressions, linear retention indices, and standard substances in PPE and PPEF, among them n-hexanal, 1-pyrroline, dimethyl trisulfide, 1-octen-3-one, 2,5-dimethyl pyrazine, 3-octen-2-one, β-damascenone, and guaiacol. The fermentation considerably amended the aroma profile of pea protein preparations resulting in a reduction or a masking of undesirable flavors.

Bitter taste receptor T2R1 is activated by dipeptides and tripeptides

Bitter taste signaling in humans is mediated by a group of 25 bitter receptors (T2Rs) that belong to the G-protein coupled receptor (GPCR) family. Previously, several bitter peptides were isolated and characterized from bitter tasting food protein derived extracts, such as pea protein and soya bean extracts. However, the molecular targets or receptors in humans for these bitter peptides were poorly characterized and least understood. In this study, we tested the ability of the bitter tasting tri- and di-peptides to activate the human bitter receptor, T2R1. In addition, we tested the ability of peptide inhibitors of the blood pressure regulatory protein, angiotensin converting enzyme (ACE) to activate T2R1. Using a heterologous expression system, T2R1 gene was transiently expressed in C6-glioma cells and changes in intracellular calcium was measured following addition of the peptides. We found that the bitter tasting tri-peptides are more potent in activating T2R1 than the di-peptides tested. Among the peptides examined, the bitter tri-peptide Phe-Phe-Phe (FFF), is the most potent in activating T2R1 with an EC50 value in the micromolar range. Furthermore, to elucidate the potential ligand binding pocket of T2R1 we used homology molecular modeling. The molecular models showed that the bitter peptides bind within the same binding pocket on the receptor. The ligand binding pocket in T2R1 is present on the extracellular surface of the receptor, and is formed by the transmembrane helices 1, 2, 3 and 7 and with extracellular loops 1 and 2 forming a cap like structure on the binding pocket.

An enzyme-based extraction process for the purification and enrichment of vegetable proteins to be applied in bakery products

In the current study an efficient aqueous extraction procedure using glucoamylase was developed to produce food grade protein extracts from lentils and white beans. The extracts contained 50.3 ± 0.6 and 49.2 ± 0.05 g protein per 100 g dry extract, respectively. The nutritional value of lentil and white bean protein extracts proved to be satisfying according to the amino acid composition, where methionine was the limiting amino acid. The produced extracts were tested for their technological properties. Both lentil and white bean protein extracts showed similar or even better technological properties regarding foaming and emulsifying capacities, heat stability and gelling properties compared to soy and pea protein extracts. Food application tests in white bread, pound cake and sponge cake proved lentil and white bean protein extracts to be potential alternatives to soy protein in replacing proteins from animal origin.

Immunoreactive properties of pea protein extract and its trypsin hydrolysates

Immunoreactive properties of pea protein extract and its trypsin hydrolysates

Alternatives to Isinglass for Beer Clarification

Fish products have been identified as serious allergens and people who are allergic to fish must avoid consuming foods made with fish or fish products. There is no evidence that isinglass (which is made from the swim bladders of certain fish) can cause allergenic reactions in people who are allergic to fish, and the European Food Safety Authority (EFSA) has ruled that it is not very likely to do so. However, many people may still wish to avoid drinking beer which has been clarified with isinglass. Two alternative compounds (which had shown promise in earlier experimental trials) for the clarification of cask ales and brewery conditioned beers were investigated. These were avian collagen and a pea protein extract. Both materials were observed to have a good clarification effect in laboratory scale fining trials and performed well in comparison to isinglass. Brewery trials were conducted using isinglass, avian collagen and pea extract as fining agents and sensory evaluation showed that there were few discernable differences between all three beers. The collagen alternative was ideal in that the data obtained from the laboratory, sensory and analytical trials show that it acted in a similar manner to isinglass and has no significant impact on beer quality. However, it was not thought that brewers would be receptive to using animal products in their beer. The pea extract also worked well and would make an excellent vegetarian alternative to isinglass, however further work is needed to scale up the production of the pea extract in order that it should become an economic option for use in the beer and wine industries.

Genetic variation in pea seed globulin composition.

A quantitative characterization of seeds from 59 pea (Pisum sativum L.) lines and relative taxa with various external characteristics and wide geographical origin was performed to explore the genetic variation of pea concerning its starch and protein contents and globulin composition. Pea lines, which produce round, wrinkled, flat, and round-dimpled seeds, have starch as the major reserve, with an average content of 46%. Protein content varied from 13.7 to 30.7% of the seed dry matter, with an overall average of 22.3%. Densitometric quantification of the individual globulins (legumin, vicilin, convicilin, and globulin-related proteins) based on SDS-PAGE gels showed no lines lacking any particular globulin. Among the lines tested, variation was shown in both their total globulins content and their globulin composition. The total globulin content ranged from 49.2 to 81.8% of the total pea protein extract (TPPE). Legumin content varied between 5.9 and 24.5% of the TPPE. Vicilin was the most abundant protein of pea, and its content varied between 26.3 and 52.0% of the TPPE. Both processed and nonprocessed vicilins occurred. The processed vicilin was the predominant one, with values between 17.8 and 40.8%, whereas the nonprocessed ones constituted between 3.1 and 13.5% of the TPPE. Convicilin was the least abundant globulin, and its content ranged from 3.9 to 8.3%. Finally, the globulin-related proteins were present in amounts ranging from 2.8 to 17.3%. They were less abundant in comparison with legumin and vicilin, but they showed the largest relative variation of the four globulin classes. Correlations between the different external characteristics and globulin composition were determined. Comparison with soybean showed that pea lines show more variety in the abundance of globulin proteins, enabling a wider range of food application.

The effect of repellents on penetration into packaging by stored-product insects

Two known repellents of stored-product insects, DEET and neem, were compared to protein-enriched pea flour, defatted protein-enriched pea flour, and pea protein extract for their efficacy at reducing penetration and invasion by several common stored-product insects: Sitophilus oryzae (L.), Tribolium castaneum (Herbst) , Cryptolestes ferrugineus (Stephens), and Oryzaephilus surinamensis (L.). The methods of preparation of pea extract affected the penetration of S. oryzae. Protein-enriched pea flour, DEET and neem reduced the penetration of S. oryzae, but defatted protein-enriched pea flour and pea protein extract did not. The number of S. oryzae, T. castaneum, C. ferrugineus and O. surinamensis entering pierced paper envelopes that contained wheat and were treated with DEET was reduced by 99%, 86%, 97% and 91%, respectively. Neem was less effective than DEET in reducing penetration and invasion of insects. Protein-enriched pea flour did not prevent insects entering pierced envelopes.

The fuc1 gene product (20 kDa FUC1) of Pisum sativum has no alpha-L-fucosidase activity.

An alpha-L-fucosidase purified from pea (Pisum sativum L. cv Alaska) epicotyl was previously described as a cell wall enzyme of 20 kDa that hydrolyses terminal alpha-L-fucosidic linkages from oligosaccharide fragments of xyloglucan. cDNA and genomic copies were further isolated and sequenced. The predicted product of the cDNA and the genomic clone (fuc1), was a 20 kDa protein containing a signal peptide and five cysteines. This was the first alpha-L-fucosidase gene to be cloned in plants but its fucosidase activity has not been demonstrated. Here, our biochemical and immuno analyses suggest that fuc1 does not encode an alpha-L-fucosidase. Pea fuc1 expressed in Escherichia coli, insect cells and Arabidopsis thaliana produced recombinant proteins without alpha-L-fucosidase activity. Pea plants had endogenous alpha-L-fucosidase activity, but the enzyme was not recognised by an antibody produced against recombinant FUC1 protein expressed in E. coli. In contrast, the antibody immunoprecipitated a 20 kDa protein which was inactive. By chromatographic analysis of pea protein extracts, we separated alpha-L-fucosidase-active fractions from the 20 kDa protein fractions. We conclude that the alpha-L-fucosidase activity is not attributable to the 20 kDa FUC1 protein. A new function for fuc1 gene product, now named PIP20 (for protease inhibitor from pea) is proposed.

Cloning and functional expression of AtCOQ3, the Arabidopsis homologue of the yeast COQ3 gene, encoding a methyltransferase from plant mitochondria involved in ubiquinone biosynthesis

A mutant of Saccharomyces cerevisiae deleted for the COQ3 gene was constructed. COQ3 encodes a 3,4-dihydroxy-5-hexaprenylbenzoate (DHHB) methyltransferase that catalyses the fourth step in the biosynthesis of ubiquinone from p-hydroxybenzoic acid. A full length cDNA encoding a homologue of DHHB-methyltransferase was cloned from an Arabidopsis thaliana cDNA library by functional complementation of a yeast coq3 deletion mutant. The Arabidopsis thaliana cDNA (AtCOQ3) was able to restore the respiration ability and ubiquinone synthesis of the mutant. The product of the 1372 bp cDNA contained 322 amino acids and had a molecular mass of 35,360 Da. The predicted amino acid sequence contained all consensus regions for S-adenosyl methionine methyltransferases and presented 26% identity with Saccharomyces cerevisiae DHHB-methyltransferase and 38% identity with the rat protein, as well as with a bacterial (Escherichia coli and Salmonella typhimurium) methyltransferase encoded by the UBIG gene. Southern analysis showed that the Arabidopsis thaliana enzyme was encoded by a single nuclear gene. The NH2-terminal part of the cDNA product contained features consistent with a putative mitochondrial transit sequence. The cDNA in Escherichia coli was overexpressed and antibodies were raised against the recombinant protein. Western blot analysis of Arabidopsis thaliana and pea protein extracts indicated that the AtCOQ3 gene product is localized within mitochondrial membranes. This result suggests that at least this step of ubiquinone synthesis takes place in mitochondria.

Pea and lentil protein extraction and functionality

These studies have demonstrated that peas and lentils can be used as protein sources for flours, concentrates and isolates. Less research attention has been devoted to lentil protein extraction, probably because of the greater cost of lentils as compared to peas. Pin‐milling and air classification is well adapted to extracting pea flours to produce pea protein concentrates. Apparently, air‐classification can be applied successfully to starch rich legumes, but will not give satisfactory results with lipid rich legumes.Wet processes, including alkaline and salt and acid solubilization, together with isoelectric precipitation or ultrafiltration, have been developed. The pea and lentil protein extracts of these processes exhibit comparable and complementary functionality to homologous soybean products. Air‐classified pea protein concentrates are different from soy protein concentrates because of residual starch which can be useful for particular functional applications. Pea isolates appear to exhibit better foaming properties and more solubility than soy isolates, but pea isolates have to be more concentrated than soy isolates to produce viscous dispersions. The economic feasibility of pea and lentil protein extracts is related directly to protein content of the flour, unique functionality of the extracts, marketability of the by‐products of extraction and the cost of peas or lentils.