Mung Bean Protein Hydrolysates Research Articles

Assessment of the DPP‐IV inhibitory potential of mung bean and adzuki bean protein hydrolysates using enzymatic hydrolysis process: specificity of peptidases and novel peptides

SummaryMung bean and adzuki bean protein concentrates were hydrolysed to the same degree of hydrolysis by two peptidases: Flavourzyme and Alcalase. Peptide fractions were analysed using MALDI‐TOF MS to investigate the differences in enzyme cleavage specificity and the inhibition activity of Dipeptidyl peptidase‐IV (DPP‐IV), using bioinformatics tools. It was shown that each peptidase showed specific selectivity in cleaving the designated substrate at a specific site. Alcalase main cutting sites on mung bean and adzuki bean proteins were identified as hydrophobic amino acid residues, particularly between Leu‐X (X‐Leu) or Phe‐X (X‐Phe) while Flavourzyme cleavage occurred between peptide bonds between Gln‐X (X‐Gln) or Pro‐X (X‐Pro). The findings demonstrated that Flavourzyme had a higher capacity for generating active peptides from both mung bean and adzuki bean protein concentrates with higher levels of DPP‐IV inhibitory activity. Proteomics and bioinformatics methodologies provided the essential knowledge required to explore novel peptides with DPP‐IV inhibitor potential from mung bean and adzuki bean hydrolysates.

Comparative study on structural, biological and functional activities of hydrolysates from Adzuki bean (Vigna angularis) and mung bean (Vigna radiata) protein concentrates using Alcalase and Flavourzyme

The physicochemical features of mung bean protein (MBP) and adzuki bean protein (ABP) hydrolysates derived from Alcalase (MBPHA, ABPHA) and Flavourzyme (MBPHF, ABPHF) were assessed using FTIR, hydrophobicity, emulsion activity, zeta potential, and health-promoting activities. The results proved that the choice of peptidase and substrate both have a significant effect on the hydrolysates in different physicochemical, structural and functional properties. Size exclusion-HPLC was used to fractionate the MBP and ABP hydrolysates. The results demonstrated that Alcalase hydrolysates included smaller peptides than Flavourzyme hydrolysates, and the chromatogram patterns of the two peptidases were similar. The peptides with the most potent antioxidant and ACE-inhibitory properties were identified using MALDI-TOF-MS. The fraction (F4) of MBPHA exhibited the highest levels of metal chelating activity. The Flavourzyme hydrolysates fraction (F2) and the ABPHA fraction (F2) showed the highest ABTS radical scavenging activity and ACE-inhibitory activity, respectively. Pro-Pro was identified in peptide sequences with ABTS radical scavenging activity as an active component while Pro-Gln was identified in peptide sequences with ACE-inhibitory activity. As a result, Pro-Pro and Pro-Gln, respectively, are likely-one of the characteristics of antioxidant and ACE-inhibitory peptides from MBP and ABP. Compared to mung bean and adzuki bean protein as substrate, Alcalase and Flavourzyme as peptidases significant impacted the development of distinct functionalities and biological activities.

Preparation, characterization and in vitro stability of iron-chelating peptides from mung beans

Mung bean protein was enzymatically hydrolyzed with either alcalase, neutral protease, or papain. The mung bean protein hydrolysates (MPH) showed good ability to chelate ferrous ions, and the chelates had high stability in vitro. The hydrolysates prepared by alcalase showed the highest degree of hydrolysis and the highest ferrous chelating rate. Single factor tests showed that the pH and the material ratio had significant effects on ferrous chelating rates. The optimal MPH to FeCl2·4H2O material ratio was 8:1 (w/w) and the optimal pH of the reaction was 7.0, which yielded a chelating rate of 96.19 ± 0.94%. The fraction 3 with the highest ferrous chelating activity up to 61.25 ± 1.02 μg/mg was obtained from MPH by affinity chromatography. Meanwhile, the MPH-Fe complex had higher digestive stability than just MPH in both in vitro and acid-alkali tolerance assays. The characterization results showed that ferrous ions mainly combined with the amino, carboxyl, imidazole and other chelating active groups in mung bean peptides to form peptide-iron chelates. Scanning electron microscopy (SEM) analysis showed that mung bean peptide chelated ferrous ions to form polymer particles. These results provided insight into ways to develop functional foods such as iron-fortified cereals.

Comparative assessment of physicochemical and antioxidative properties of mung bean protein hydrolysates.

Two commercial plant proteases namely ficin and bromelain, were acquired to hydrolyze mung bean protein over 300 min hydrolysis, and the physicochemical and antioxidative properties of the obtained hydrolysates were investigated. Bromelain-treated mung bean protein hydrolysates presented a higher degree of hydrolysis in comparison with ficin-treated hydrolysates, further modifying their physicochemical and emulsifying properties. All mung bean protein hydrolysates exhibited 50% scavenging of DPPH radical (IC50) in the concentration range from 8.67 to 16.22 μg mL−1. Our results also showed that strong metal ion-chelating activity was found in the ficin- (higher activity) and bromelain-treated protein hydrolysates. In addition, oxidative stability of linoleic acid was significantly enhanced by two selected protein hydrolysates, particularly the bromelain-treated hydrolysate with the highest inhibition effect of linoleic acid oxidation (94.55 ± 0.10%). Interestingly, both of these two hydrolysates could effectively retard lipid oxidation of sunflower oil and sunflower oil-in-water emulsion, while the ficin-treated hydrolysate showed slightly better performance. Therefore, mung bean protein hydrolysates showed potential to inhibit lipid oxidation, which could be advantageous in the food industry for producing fortified food.

Mung Bean Protein Hydrolysates Protect Mouse Liver Cell Line Nctc-1469 Cell from Hydrogen Peroxide-Induced Cell Injury.

Mung bean is nutritious and rich in protein (19.5%–33.1%). However, there are few studies on mung bean protein active peptides so the mung bean protein hydrolysates (MBPHs) were investigated for evaluating their ability to clear intracellular reactive oxygen species (ROS) and regulating the ability of antioxidant enzymes on NCTC-1469 cells. Results showed that MBPHs, MBPHs-I (molecular weight < 3 kDa), MBPHs-II (molecular weight between 3 and 10 kDa), and MBPHs-III (molecular weight > 10 kDa) could all improve the survival rate of cells compared with the model group. MBPHs, MBPHs-I, and MBPHs-II could significantly decrease the content of lactate dehydrogenase (LDH) and reduce the generation of malonaldehyde (MDA) at a concentration of 0.4 mg/mL. Regarding the intracellular ROS, the result showed that MBPHs-I significantly reduced the production of ROS (from 58.3% to 26.6%) and had a dose-dependent relationship. In addition, the amino acid analysis showed that MBPHs-I had a balanced amino acid composition. MBPHs-I is rich in lysine but was deficient in cereals. Therefore, the hydrophobic and aromatic amino acids in MBPHs-I were high, which could improve its antioxidant activity. According to the results, MBPHs-I was the best and most potent natural antioxidant and it can contribute to drug development and medical application.

Isolation and characterization of zinc-binding peptides from mung bean protein hydrolysates

The aim of this study was to isolate, purify, and characterize a novel peptide with zinc-binding ability from mung bean protein hydrolysates obtained by alcalase. The peptide purification process was performed using ultrafiltration, TSK-GEL size-exclusion chromatography, and reversed-phase high-performance liquid chromatography (RP-HPLC). The molecular weight of the isolated peptide was determined by liquid chromatography–electrospray ionization mass spectrometry (LC–ESI/MS) and found to be 1192.56 Da with amino acid sequence SSEDQPFNLR. The identified peptide showed zinc-binding capacity of 80.82 mg/g, which is 55.09% higher than that of the original mung bean protein hydrolyzate. Moreover, UV–vis and FTIR spectra indicated that the zinc-binding sites on the peptide were nitrogen atoms of amide, terminal amino group, and oxygen atoms on the carboxyl group. Moreover, zinc-releasing experiments showed that the peptide–zinc chelate is highly soluble under both acidic and alkaline conditions. Furthermore, the identified peptide significantly promoted the transportation and absorption of zinc ions in Caco-2 cell model. These findings indicate a potential for production of zinc-binding peptide from mung bean protein for the utilization as ingredient in functional foods.

Finding and isolation of novel peptides with anti-proliferation ability of hepatocellular carcinoma cells from mung bean protein hydrolysates

Abstract Mung bean protein hydrolysate has shown some biological properties. Isolated mung bean proteins were hydrolyzed by papain and tested for antiproliferative effects on cancer cells. It was confirmed that the mung bean peptides could inhibit the proliferation of tumor cells in mice, without the side effects of chemical anticancer drugs, and that the mung bean peptides promoted apoptosis of HepG2 cells and arrested the cell cycle in the S phase at a low dose and the G0/G1 phase at a high dose. Subsequently, the hydrolysates were purified through ultrafiltration and Sephadex G-15 gel chromatography. Fraction A, obtained following size-fractionation (

Anti-allergic activity of mung bean (Vigna radiata (L.) Wilczek) protein hydrolysates produced by enzymatic hydrolysis using non- gastrointestinal and gastrointestinal enzymes

Mung bean seed is a well-known plant protein consumed in Asian countries but the protein is usually retrieved as a waste product during starch production. This study investigated the anti-allergic property of mung bean protein hydrolysates (MBPH) produced by enzymatic hydrolysis using non-gastrointestinal (non-GI), GI and a combination of non-GI+GI enzymes. The hydrolysates were investigated for any anti-allergic property by detecting the amount of β-hexosaminidase released in RBL-2H3 cells, and complemented with the MTT assay to show cell viability. It was found that MBPH hydrolyzed by a combination of flavourzyme (non-GI enzyme) and pancreatin (GI enzyme) exhibited the highest anti-allergic activity (135.61%), followed by those produced with alcalase, a non-GI enzyme (121.74%) and 80.32% for pancreatin (GI enzyme). Minimal toxicity (<30%) of all hydrolysates on RBL-2H3 cells line was observed. The results suggest that MBPH can potentially serve as a hypoallergenic food ingredient or supplement. PRACTICAL APPLICATIONS: Mung bean (Vigna radiata L. (Wilczek)) is also known as "green gram" and it is an excellent source of protein. The major mung bean storage proteins are the globulin, albumin and legumin, which are also referred to as legume allergens. Our study showed that mung bean peptides obtained after enzymatic hydrolysis influenced β-hexosaminidase inhibition without any toxic effect on RBL-2H3 cells. This indicates that mung bean allergenicity can be reduced after enzymatic hydrolysis and the protein hydrolysates could be as a hypoallergic food, ingredient, supplement and/or protein substitute in the formulation of food products.

Structural and functional characterization of calcium and iron-binding peptides from mung bean protein hydrolysate

This work was aimed at producing peptides containing calcium and iron-binding capabilities from mung bean protein extract after enzymatic hydrolysis. Mung bean protein hydrolysates were separated by ultrafiltration into eight fractions, and assayed for calcium and iron-binding properties. The <5 kDa fraction was the most active and was separated by size exclusion chromatography into 4 fractions. Fraction 2 had the most active calcium and iron-binding activities, and further identified by mass spectrometry coupled with FindPept tool. Potential peptides sequences (133) were identified and 10 with high contents of leucine, isoleucine and aspartic acid were synthesized. Peptides LLLGI, AIVIL and HADAD were the best calcium binders while PAIDL was the most potent iron-binding peptide. The presence of L or I at the C- or N-terminal but not together at both terminals may have contributed to better calcium-binding ability. These peptides are potential ingredients to formulate functional foods with enhanced mineral-binding properties.

Determination of Nutritional and Bioactive Properties of Peptides in Enzymatic Pea, Chickpea, and Mung Bean Protein Hydrolysates

Within the primary structure of many pea and mung bean proteins are peptide sequences that can potentially be used in the formulation of therapeutic products for the treatment and prevention of human diseases. However, these peptide sequences need protease treatments before they can be released free of the parent proteins. Unlike chemical hydrolysis, enzymatic treatment enables more efficient tailoring of peptide products without formation of toxic by-products or destruction of amino acids. This review provides information on current methods that have been used to convert inactive pea and mung bean proteins into bioactive peptides. It focuses on 3 main bioactive properties, such as inhibitions of (1) angiotensin converting enzyme (ACE) activity; (2) calmodulin (CaM)-dependent enzymes; and (3) copper-chelating activity. ACE is an established marker for hypertension, high levels of some CaM-dependent enzymes are risk factors for various human diseases including cancer and Alzheimer's disease, and high vascular copper concentrations may potentiate atherosclerosis. Also reviewed are the production and evaluation of activity of hypoallergenic peptides that may offer protection against anaphylactic reactions. The 3 main proteins discussed are chickpea, mung bean, and field pea.

Antihypertensive effect of alcalase generated mung bean protein hydrolysates in spontaneously hypertensive rats

Mung bean protein isolates were hydrolyzed by alcalase and neutrase, respectively, and the angiotensin I-converting enzyme (ACE) inhibitory activities of the enzymatic hydrolysates were determined at different hydrolysis times. The highest ACE inhibitory activity recorded was for a hydrolysate generated by alcalase (IC50: 0.64 mg protein/ml) after 2 h of hydrolysis. A significant decrease in systolic blood pressure was observed in spontaneously hypertensive rats following a single oral administration of this hydrolysate at a dose of 600 mg/kg of body weight.

Mung-bean Protein Hydrolysates Obtained with Alcalase Exhibit Angiotensin I-converting Enzyme Inhibitory Activity

Mung-bean protein isolates were hydrolysed by two proteases alcalase and neutrase commercially available for food industry use, and the angiotensin I-converting enzyme (ACE) inhibitory activities of the enzymatic hydrolysates were measured at different hydrolysis times. The non-hydrolysed protein showed no inhibitory activity. Hydrolysates generated with neutrase displayed very low ACE inhibitory activity, while those obtained with alcalase exhibited high inhibitory activity. The highest ACE inhibitory activity with the IC50 value of 0.64 mg protein/mL was found in the hydrolysate obtained with alcalase at 2h of hydrolysis time. These results indicated that mung-bean protein is a good protein source of ACE inhibitory peptides when hydrolysed with the protease alcalase. The mung-bean protein hydrolysates prepared with alcalase might be utilised for physiologically functional foods with antihypertensive activity.