Добірка наукової літератури з теми "Pea protein hydrolysates"

Abstract Protein hydrolysate contains a mixture of various lengths of short peptides chain and free amino acids that may excert biological activities. This research aims to screen potential antioxidant and antibacterial activities of protein hydrolysate produced from three kinds of germinated beans i.e. lablab bean (Lablab purpureus), pigeon pea (Cajanus cajan (L.) Millsp) and kidney bean (Phaseolus vulgaris) through enzymatic hydrolysis process. The steps of research included germination process of the beans prior to total protein isolation, enzymatic hydrolysis of total protein isolates using pancreatin enzyme, evaluation of in vitro antioxidant activity of the hydrolysates protein using DPPH (1,1-diphenyl-2-picryl hydrazyl) method, and antibaterial activity testing towards Eschericia coli and Staphyllococcus aureus bacteria. The results revealed that pancreatine enzyme was able to hydrolyse germinated protein of lablab bean, pigeon pea and kidney bean at the experiment condition applied with degree of hydrolysis 34.12%, 27.44%, and 30,93% respectively. It was also found that protein hydrolysates of lablab bean, pigeon pea, and kidney bean demonstrated antioxidant activity which percentage radical DPPH scavenging activity of 84.02%, 68.97% and 67.89 %. On the other hand, all of those protein hydrolysates did not show any antibacterial activity towards Eschericia coli and Staphyllococcus aureus bacteria.

Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea (Pisum sativum L.) seed meal proteins (albumins, legumins and vicilins) were isolated, submitted to a simulated gastrointestinal digestion, and the effects of their hydrolysates (pea albumins hydrolysates (PAH), pea legumins hydrolysates (PLH) and pea vicilin hydrolysates (PVH), respectively) on 3T3-L1 murine pre-adipocytes were investigated. The pea vicilin hydrolysate (PVH), but not native pea vicilins, increased lipid accumulation during adipocyte differentiation. PVH also increased the mRNA expression levels of the adipocyte fatty acid-binding protein (aP2) and decreased that of pre-adipocyte factor-1 (Pref-1) (a pre-adipocyte marker gene), suggesting that PVH promotes adipocyte differentiation. Moreover, PVH induced adiponectin and insulin-responsive glucose transporter 4 (GLUT4) and stimulated glucose uptake. The expression levels of peroxisome proliferator-activated receptor γ (PPARγ), a key regulator of adipocyte differentiation, were up-regulated in 3T3-L1 cells treated with PVH during adipocyte differentiation. Finally, PVH exhibited PPARγ ligand activity. Lactalbumin or other pea hydrolysates (PAH, PLH) did not exhibit such effects. These findings show that PVH stimulates adipocyte differentiation via, at least in part, the up-regulation of PPARγ expression levels and ligand activity. These effects of PVH might be relevant in the context of the beneficial health effects of legume consumption in obesity-associated metabolic disorders.

In this work, we report the potency of enzymatic hydrolysates of pea proteins against trypsin and chymotrypsin. Pea protein concentrate was digested with each of alcalase, chymotrypsin, pepsin, and trypsin, followed by membrane separation of the protein hydrolysates into peptide fractions (<1, 1–3, 3–5, and 5–10 kDa). Peptide size profiling with size-exclusion gel chromatography indicated the narrowest size range (0.85–4.98 kDa) for alcalase. Trypsin activity was strongly (p < 0.05) inhibited by the ultrafiltration fractions (mean IC50 = 2.2 mg/mL) obtained from the trypsin hydrolysate when compared to the unfractionated hydrolysate (IC50 = 6.8 mg/mL). Similarly, ultrafiltration also enhanced trypsin inhibition by the alcalase-digested peptides with an IC50 of 21.4 mg/mL for the unfractionated hydrolysate in comparison to 3.1–4.7 mg/mL for the fractions. However, ultrafiltration did not enhance trypsin inhibitory activity of chymotrypsin-digested peptides, while the peptide separation reduced efficacy of pepsin-digested peptides. In contrast, chymotrypsin inhibition by all the enzymatic digests was significantly (p < 0.05) enhanced by ultrafiltration, especially peptide sizes >3 kDa. Kinetics of enzyme inhibition indicate peptides were bound to the enzyme active site in a competitive mode that led to reduced catalysis. We conclude that the pea peptides could function as useful tools to promote human health and as a preservative during food processing and storage.

Pigeon pea (Cajanus cajan (L) Mill sp.) seeds are rich sources of protein in the legume family and their consumption has been associated with the prevention of noncommunicated diseases, which is attributable to their content of bioactive components. Antioxidant protein hydrolysates were produced from pigeon pea protein isolate (PPI) by enzymatic hydrolysis using pancreatin and flavourzyme. The hydrolysates were analyzed for their physicochemical, molecular weight, amino acid composition, and in vitro antioxidant activities. The molecular weights of polypeptides in the hydrolysates were 8, 20, 25 and 48 kDa, which were determined after pancreatin or flavourzyme hydrolysis of the protein isolate for 4 h. Pancreatin-hydrolyzed pigeon pea protein (PPHP) contained high hydrophobic amino acids, especially isoleucine, leucine and valine, which were related to the high content of aromatic amino acids. The hydrolysates obtained from flavourzyme hydrolysis of pigeon pea proteins (PPHF) presented significantly higher capacities to scavenge ABTS˙+ and reduce Fe3 + better than that of PPI, while the PPI exhibited strong DPPH scavenging (98.4 mg trolox equivalent antioxidant capacity). The results indicated that the partial hydrolysis for PPI provided medium to high molecular weight of peptides. Therefore, PPHF could be a promising source of bioactive peptides and a potential ingredient for the formulation of functional foods against oxidative stress

Animal protein plays a key role in the human diet as the most balanced amino acid composition; however, its consumption often causes allergic reactions. Plant protein serves as a substitute for animal protein. The most promising sources of plant protein are the seeds of cereals, pulses, oilseeds and cereals. Research aim: selection of conditions for obtaining protein isolates and enzymatic hydrolysates having the desired functional properties from different types of vegetable raw materials.Pea, corn and oat flour LLC "FavoritT"; linseed flour LLC NGO "Compass Health". Enzyme preparations: chymotrypsin LLC "Samson-Med"; Protex 40E Genencor; protosubtilin G3x produced by PO "Sibbiofarm" LLC; pancreatin PJSC "Biosintez"; trypsin LLC "Diaem"; beef pepsin OJSC "MHSF". Crude protein content was determined by Kjeldahl method, protein substances - by modified Lowry method. Fat-holding, water-holding, emulsifying and foaming capacities, as well as allergenicity of protein isolates and hydrolysates were determined. Conditions for protein substances extraction from flax, corn, oat and pea flour with the yield of high-molecular protein fraction not less than 70 % of raw protein content were selected. The conditions of protein isolates precipitation to produce preparations containing not less than 85% of protein have been selected. The type of enzyme preparation for hydrolysis - pancreatin - was selected. It was found that in order to increase water- and fat-holding capacity of pea isolate, hydrolysis is possible with duration not exceeding 15 min, for all other isolates hydrolysis is undesirable. The best emulsifying and foam-forming capacities are possessed by linseed hydrolysates after 60 and 90 min of hydrolysis, respectively. Enzymatic hydrolysis was shown to reduce the allergenicity of plant proteins. The obtained hydrolysates of vegetable proteins can be used as ingredients for functional products, as well as for obtaining products with reduced allergenicity.

Nine Lactobacillus strains known for surface proteinase activity were chosen from our collection and tested for their ability to grow in pea seed protein-based medium, and to hydrolyze purified pea proteins in order to produce peptides with antioxidant (AO) activity. Two strains, Lactobacillus rhamnosus BGT10 and Lactobacillus zeae LMG17315, exhibited strong proteolytic activity against pea proteins. The AO activity of the pea hydrolysate fraction, MW <10 kDa, obtained by the fermentation of purified pea proteins with Lactobacillus rhamnosus BGT10, was tested by standard spectrophotometric assays (DPPH, ABTS, Fe3+-reducing capacity) and the recently developed direct current (DC) polarographic assay. The low molecular weight fraction of the obtained hydrolysate was separated using ion exchange chromatography, while the AO activity of eluted fractions was determined by means of a sensitive DC polarographic assay without previous concentration of samples. Results revealed that the fraction present in low abundance that contained basic peptides possessed the highest antioxidant activity. Based on the obtained results, it can be concluded that Lactobacillus rhamnosus BGT10 should be further investigated as a candidate strain for large-scale production of bioactive peptides from legume proteins.

The objective was to investigate the anti-adipogenesis potential of selected legume protein hydrolysates (LPH) and combinations using biochemical assays and in silico predictions. Black bean, green pea, chickpea, lentil and fava bean protein isolates were hydrolyzed using alcalase (A) or pepsin/pancreatin (PP). The degree of hydrolysis ranged from 15.5% to 35.5% for A-LPH and PP-LPH, respectively. Antioxidant capacities ranged for ABTS•+ IC50 from 0.3 to 0.9 Trolox equivalents (TE) mg/mL, DPPH• IC50 from 0.7 to 13.5 TE mg/mL and nitric oxide (NO) inhibition IC50 from 0.3 to 1.3 mg/mL. LPH from PP–green pea, A–green pea and A–black bean inhibited pancreatic lipase (PL) (IC50 = 0.9 mg/mL, 2.2 mg/mL and 1.2 mg/mL, respectively) (p < 0.05). For HMG-CoA reductase (HMGR) inhibition, the LPH from A–chickpea (0.15 mg/mL), PP–lentil (1.2 mg/mL), A–green pea (1.4 mg/mL) and PP–green pea (1.5 mg/mL) were potent inhibitors. Combinations of PP–green pea + A–black bean (IC50 = 0.4 mg/mL), A–green pea + PP–green pea (IC50 = 0.9 mg/mL) and A–black bean + A–green pea (IC50 = 0.6 mg/mL) presented synergistic effects to inhibit PL. A–chickpea + PP–lentil (IC50 = 0.8 mg/mL) and PP–lentil + A–green pea (IC50 = 1.3 mg/mL) interacted additively to inhibit HMGR and synergistically in the combination of A–chickpea + PP–black bean (IC50 = 1.3 mg/mL) to block HMGR. Peptides FEDGLV and PYGVPVGVR inhibited PL and HMGR in silico, showing predicted binding energy interactions of −7.6 and −8.8 kcal/mol, respectively. Combinations of LPH from different legume protein sources could increase synergistically their anti-adipogenic potential.

Les peptides chélateurs de métaux (PCMs), issus d'hydrolysats de protéines, présentent diverses applications dans les domaines de la nutrition, de la pharmacie, des cosmétiques, etc. Cependant, l'approche empirique généralement utilisée pour découvrir des peptides bioactifs à partir d'hydrolysats est longue et coûteuse en raison de nombreuses étapes de fractionnement, de séparation et d'évaluation des activités biologiques. Cette thèse a donc pour but de développer une nouvelle approche pour la prédiction de la séparation des PCMs en utilisant la modélisation et la simulation chromatographiques basées sur l'analogie entre la chromatographie d'affinité sur ions métalliques immobilisés (IMAC) et la résonance plasmonique de surface (SPR). Pour la première fois, l'analogie SPR-IMAC a été étudiée expérimentalement sur 22 peptides et 70% d'entre eux ont validé cette analogie, puisque les peptides bien retenus en IMAC étaient également dotés d'une bonne affinité pour Ni2+ en SPR. Dans un deuxième temps, des peptides ayant une forte affinité pour Ni2+ (c'est-à-dire une faible constante de dissociation KD en SPR et un temps de rétention élevé en IMAC) ont été utilisés pour étudier la simulation des profils de concentration de peptides à la sortie de la colonne en IMAC. La connaissance des isothermes d'adsorption étant nécessaire pour effectuer la simulation, il a fallu développer une méthodologie pour prédire les paramètres de l'isotherme de Langmuir en IMAC à partir des données SPR. La simulation a été évaluée en comparant les temps de rétention expérimentaux et simulés qui devraient être proches pour une prédiction fiable. Par conséquent, plusieurs approches ont été étudiées pour déterminer les paramètres de sorption de Langmuir. L‘approche la plus intéressante a introduit un facteur correctif sur la capacité d'adsorption maximale qmax seulement. En effet, l'affinité des peptides pour le Ni2+ immobilisé étant supposée constante quelle que soit technologie utilisée (SPR vs. IMAC), la constante d'affinité KA n'a pas été modifiée. Parallèlement, les applications industrielles des PCMs et des hydrolysats peptidiques ont été étudiées. Tout d'abord, des hydrolysats de protéines de pois ont été produits par protéolyse enzymatique soit avec l'Alcalase® suivi de la Flavourzyme® (Alc+Flav≤1kDa), soit par la Protamex® suivi de la Flavourzyme® (Prot+Flav≤1kDa). La technologie SwitchSENSE® a mis en évidence la présence de peptides chélateurs de Ni2+ et les tests antioxydants ont montré que l'hydrolysat Prot+Flav≤1kDa avait une activité antiradicalaire et un pouvoir réducteur plus élevés, liés à son degré d'hydrolyse plus élevé et à la quantité de peptides de petite taille. Les hydrolysats de protéines de pois et les PCMs ont ensuite été étudiés pour leur capacité à inhiber l'oxydation des lipides dans les émulsions. Ils ont ralenti l'oxydation des lipides par chélation des métaux pro-oxydants (tels que Fe2+) en réduisant les produits d'oxydation primaires et secondaires, responsables de la détérioration des produits contenant des lipides. Ainsi, les hydrolysats de pois et les PCMs pourraient être utilisés comme antioxydants dans les produits alimentaires et cosmétiques, comme alternatives aux produits chimiques tels que l'EDTA, le BHT et le TBHQ
Metal-Chelating Peptides (MCPs), from protein hydrolysates, present various applications in nutrition, pharmacy, cosmetic etc. Yet, the empirical approach generally used to discover bioactive peptides from hydrolysates is time consuming and expensive due to many steps of fractionation, separation and biological activities evaluation. Thus, this PhD aimed to develop a novel approach for MCPs separation prediction using chromatography modelling and simulation based on the analogy between Immobilized Metal ion Affinity Chromatography (IMAC) and Surface Plasmon Resonance (SPR). For the first time, the SPR-IMAC analogy was experimentally investigated on 22 peptides and 70% of them validated this analogy, since peptides well retained in IMAC were also endowed with a good affinity for Ni2+ in SPR. In the second time, peptides with high affinity for Ni2+ (i.e low dissociation constant KD in SPR and a high retention time in IMAC) were used to study the modelling and simulation of peptide concentration profiles at the column outlet in IMAC. Since knowledge of adsorption isotherms was required to perform simulation, it was necessary to develop a methodology for predicting Langmuir isotherm parameters in IMAC from SPR data. The validity of simulation was evaluated by comparing experimental and simulated retention times that should be close for reliable prediction. Therefore, several approaches were evaluated to determine Langmuir sorption parameters, the most interesting one introduces a correction factor on the maximum adsorption capacity qmax alone, assuming that the affinity of peptides for immobilized Ni2+ did not change depending on the technology used (SPR vs. IMAC), thus affinity constant KA was not modified. Meanwhile, industrial application of MCPs and hydrolysates were studied. First, pea protein hydrolysates were produced by either Alcalase® followed by Flavourzyme® (Alc+Flav≤1kDa) or Protamex® followed by Flavourzyme® (Prot+Flav≤1kDa). SwitchSENSE® technology evidences the presence of Ni2+ chelating peptides and antioxidants tests showed that Prot+Flav≤1kDa has higher radical scavenging and reducing power, related to its higher degree of hydrolysis and small-size peptides quantity. Secondly, pea hydrolysates and MCPs were investigated for their ability to inhibit the lipid oxidation in emulsions. They slowed down lipid oxidation through chelation of prooxidant (metals such as Fe2+) reducing primary and secondary oxidation products responsible of deterioration of lipid containing products. Thus, pea hydrolysates and MCPs could be used as antioxidants in food and cosmetic products, as alternative to chemicals such as EDTA, BHT and TBHQ

The effects of heat or high pressure treatment on the physicochemical and functional properties of pea proteins were evaluated by measuring polypeptide composition, hydrophobicity, solubility, gelation, emulsification, foaming, water-holding capacity and oil-holding capacity. Heat processing (≥ 70 °C) and high pressure treatment (≥ 200 MPa) led to significant increase (from 1.41 to 2.42) in hydrophobicity of native pea proteins. Native gel electrophoresis showed that the content of 11S protein decreased (increased aggregation) as intensity of pressure treatment was increased. In contrast the 7S protein was resistant to pressure-induced protein aggregation. The solubility and emulsifying capacity of pea proteins processed under higher pressure or heat at neutral pH had slight decreases probably due to the formation of aggregates. High pressure treatment of pea proteins led to reductions in the amount of protease required to produce renin-inhibitory peptides.

Pea protein hydrolysate (PPH) has antihypertensive effects and prostanoids have been implicated in renal diseases. To investigate the role of PPH and prostanoids on renal and cardiovascular effects in cardio-renal disease, normal and diseased Han:SPRD-cy rats were given diets containing either 0, 0.5% or 1% PPH for 8 weeks. At termination, diseased rat kidneys displayed increased renal cyst growth, fibrosis, plasma creatinine and lower monocyte chemoattractant protein-1. Diseased rats also exhibited left ventricular (LV) hypertrophy, elevated systolic and diastolic blood pressures and LV end diastolic and systolic pressures. Four of five prostanoids were elevated in diseased rat kidneys. PPH attenuated systolic blood pressure, but not other components of the cardio-renal syndrome. PPH also increased select prostanoids in normal and diseased rats. Thus, dietary PPH attenuates hypertension in the Han:SPRD-cy rat, but does not ameliorate other components of disease, possibly due to increased prostanoid effects or an insufficient treatment length.

Overview: Alzheimer’s disease (AD) is a neurodegenerative disorder prevalent among the aged population with morbidity and mortality rate of ~ 12%. Research has linked the cause of this disorder to the loss of acetylcholine through excessive activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Therefore, a promising therapeutic approach for AD treatment is the inhibition of AChE/BChE activities. Common features of an AD brain include low levels of acetylcholine, the presence of amyloid-β peptides deposits and severe oxidative stress triggered by lipid peroxidation and formation of free radicals. Natural peptides that possess antioxidant and bioactivities could have prospects for use in AD management. Objectives: Optimize enzymatic hydrolysis of yellow field pea proteins into protein hydrolysates that possess antioxidant, anti-AChE and anti-BChE activities. Methods: Pea protein (70%) was hydrolyzed using six as alcalase (AH), chymotrypsin (CHH), flavourzyme (FZH), pancreatin (PCH), pepsin (PEH) and trypsin (TPH). The supernatants were sequentially passed through ultrafiltration (UF) membranes with molecular weight cut-off of 1, 3, 5 and 10 kDa to collect the permeates as < 1, 1-3, 3-5, and 5-10 kDa, respectively. The hydrolysates and UF fractions were screened for inhibition of linoleic acid peroxidation (LAP) in addition to radical scavenging (hydroxyl and superoxide) and anti-AChE/BChE properties.Results: Hydrolysates showed varying degrees of radicals scavenging and LAP, as well as anti-AChE and anti-BChE activities but the potency improved by >10% for most UF fractions. AH, FZH, PEH and the UF fractions (1-3 kDa) exhibited better and statistically significant (p<0.05) radical scavenging and AChE/BChE activities than other hydrolysates by 20-30% and 20-40% respectively at same concentrations (10-50 µg).Significance of study: The results suggest that pea protein-derived peptides could be potential candidates for use in the inhibition of AChE and BChE activities, which could provide therapeutic tools suitable for the prevention and treatment of AD.

The fish and shellfish industry processes 851,984 tonnes of fish per year worldwide. However, only 43% of that is consumed, and valuable proteins are processed as waste. Protein hydrolysates are widely used in food technology for their nutritional and functional properties. The goal of this project is to extract protein from whelk by-products derived from the shellfish processing industry and create protein hydrolysates that have marketable value. The by-products were divided into two types: raw (R) and cooked byproduct (C). The proteins were extracted using the pH shift method and quantified using the Bradford assay. It was possible to extract a maximum of 455 mg/g at a neutral pH, for which R had the highest protein yield. Proteins were also qualified using reverse phase high-performance liquid chromatography (RP-HPLC) that showed that R has more hydrophilic proteins while the C extracted protein showed more peaks in the hydrophobic phase. The Fourier-transform infrared spectroscopy (FTIR) indicated the presence of glutamine, tyrosine, and serine in the extracted proteins. Extracted proteins were then hydrolyzed using Alcalase and α-Chymotrypsin. It was possible to obtain higher degrees of hydrolysis (DH) using Alcalase. The hydrolysates were tested for antioxidant activity using the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical antioxidant assay. Alcalase hydrolysates showed to have overall lower IC50 for stabilization of the DPPH radical than α-Chymotrypsin, the lowest one being 13.92±1.57 µg/mL for the Alcalase hydrolyzed neutral proteins. The IC50 results obtained are significantly lower than the ones described in other studies using the same enzymes or other marine species. This can indicate that more heterogenous mixtures of by-product can originate extracted proteins that when hydrolyzed lead to higher radical scavenging activity, thus making shellfish industry by-product a sustainable and valuable source of antioxidant peptides. Keywords: Shellfish; Bioactive peptides; Protein extraction; Protein hydrolysates, Waste management, Nutraceuticals

Slaughterhouse blood is a valuable by-product since multiple bioactive compounds can be derived out of it. Its solid fraction consists mainly of hemoglobin, which is a good source of antimicrobial and antioxidant peptides that can be released by peptic hydrolysis. Nevertheless, this method has limitations such as low yield, expensive cost of enzyme process, and non-eco-friendly production (high energy consumption and chemical reagents requested). Amount the alternative green technologies for protein valorization, pulsed electric field (PEF) stands out since it allows modifying the physicochemical properties of proteins, promoting the enzymatic hydrolysis, enzyme inactivation, and bioactivity enhancement. Thus, this study aimed to evaluate the effect of PEF on the pepsin inactivation and biological activities (antimicrobial and antioxidant) in hemoglobin hydrolysates. Bovine and porcine hemoglobins were hydrolyzed with pepsin for 3 h (37°C, pH 3.0) and treated with PEF (73 pulses, 23.8kV/cm, 90Hz) to inactivate the enzyme. The hydrolysis degree was evaluated, which did not show significant changes after PEF-inactivation of pepsin, whereas the peptide population analysis by RP-UPLC-MS/MS showed some changes in PEF-treated hydrolysates over time, which suggested a residual pepsin activity. Additionally, the impact of pH (3, 7, and 10) on bioactivity was studied. PEF-treatments did not show a significant impact on antimicrobial (antibacterial, antifungal, and anti-yeast activities) and antioxidant activities (DPPH and ORAC). However, higher pH fostered stronger anti-yeast activity (R. mucilaginosa) and DPPH‐scavenging capacity, whereas pH 7 fostered the antifungal activity (M. racemosus). Even though some changes were observed in the peptide population, no negative effects of PEF were found for biological activities. Thus, the utilization of hemoglobin from the meat industry combined with PEF-treatment fits the circular economy concept since derived peptides can be recycled to protect meat and other products against microbial growth and oxidation.