Antioxidant Potential of Mung Bean (Vigna radiata) Albumin Peptides Produced by Enzymatic Hydrolysis Analyzed by Biochemical and In Silico Methods
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Mung Bean Albumin Extraction
2.3. Gel Electrophoresis
2.3.1. Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis (SDS-PAGE)
2.3.2. Native Blue–Polyacrylamide Gel Electrophoresis (Native Blue-PAGE)
2.4. In Silico Hydrolysis of Mung Bean Albumin Sequences
2.5. Enzymatic Hydrolysis
2.6. Antioxidant Assays
2.6.1. ABTS Radical Scavenging Assay
2.6.2. Ferrous Ion Chelation Assay
2.6.3. Oxygen Radical Absorbance Capacity (ORAC) Assay
2.7. Peptide Sequencing
2.8. Statistical Analysis
3. Results
3.1. Mung Bean Albumin Profiles
3.2. Potentially Antioxidant Peptides from Mung Bean Albumins by In Silico Hydrolysis
3.3. Antioxidant Activity of Mung Bean Albumin Hydrolysates and Peptide Sequencing
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Protease | P4 Position | P3 Position | P2 Position | P1 Position | P1′ Position | P2′ Position | P3′ Position | P4′ Position |
---|---|---|---|---|---|---|---|---|
Alcalase | Gly, Pro, Ala, Val, Leu, Met, Phe, Tyr, Ser, Thr, Asn, Glu, His | Gly, Ala, Val, Leu, Phe, Ser, Thr, Asn, Gln, Asp, Glu, Lys, Arg, His | Gly, Pro, Ala, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Asn, Gln, Glu, Arg, His | Ala, Val, Leu, Met, Phe, Tyr, Ser, Asn, Gln, Glu, Lys, Arg, His | Gly. Ala, Val, Leu, Met, Phe, Tyr, Ser, Thr, Gln, His | Gly, Pro, Ala, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Gln, Glu, Arg, His | Gly, Pro, Ala, Val, Leu, Met, Phe, Tyr, Ser, Thr, Asn, Lys, Arg, His | Gly, Pro, Ala, Val, Leu, Tyr, Ser, Thr, Cys, Asn, Gln, Asp, Glu, Lys, His |
Stem Bromelain | Pro, Ala, Leu, Ile, Phe, Tyr, Ser, Thr, Cys, Glu, Arg, His | Gly, Pro, Ala, Val, Leu, Phe, Ser, Thr, Lys, Arg, His | Gly, Pro, Val, Leu, Phe, Tyr, Ser, Thr, Asn, Arg, His | Gly, Ala, Val, Leu, Phe, Tyr, Ser, Thr, Asn, Gln, Arg | Gly, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Asn, Gln, Asp, Glu, Lys, His | Pro, Ala, Val, Leu, Tyr, Ser, Thr, Asn, Gln, Asp, Glu, Lys, His | Gly, Pro, Val, Leu, Phe, Ser, Thr, Cys, Asn, Gln, Asp, Glu, Arg, His | Gly, Pro, Ala, Val, Leu, Phe, Tyr, Ser, Thr, Asp, Glu, Lys, Arg, His |
Ficin | Pro, Leu, Ser, Glu, Lys, Arg, His | Gly, Ala, Val, Leu, Ile, Phe, Thr, Arg, His | Gly. Val, Leu, Phe, Thr, Lys | Gly, Leu, Phe, Tyr, Ser, Lys, Arg, His | Leu, Phe, Tyr, Ser, Thr, Lys, His | Val, Leu, Tyr, Thr, Asn, Lys, His | Pro, Ala, Val, Leu, Ser, Thr, Glu, Lys | Gly, Pro, Val, Asn, Asp, Glu, Lys |
Papain | Gly, Pro, Ala, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Cys, Asn, Asp, Glu, Arg, His | Gly, Pro, Ala, Val, Leu, Ile, Met, Phe, Tyr, Ser, Gln, Asp, Glu, Lys, Arg, His | All 20 Canonical AA | All 20 Canonical AA | Gly, Ala, Val, Leu, Ile, Met, Phe, Tyr, Ser, Thr, Asn, Gln, Asp, Glu, Lys, His | Gly, Pro, Ala, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Cys, Asn, Gln, Glu, Arg, His | Gly, Pro, Ala, Val, Leu, Phe, Tyr, Ser, Cys, Glu, Lys, Arg, His | Gly, Pro, Ala, Val, Leu, Ile, Phe, Tyr, Ser, Thr, Asn, Asp, Glu, Lys, Arg, His |
Thermolysin | All 20 Canonical AA | All 20 Canonical AA | All 20 Canonical AA | All 20 Canonical AA | Gly, Pro, Ala, Val, Leu, Ile, Met, Phe, Tyr, Trp, Ser, Thr, Asn, Gln, Asp, Glu, Lys, His | All 20 Canonical AA | All 20 Canonical AA | All 20 Canonical AA |
Protease | Protein Sequence | Potentially Antioxidant Di-/Tri-Peptides Expected to be Produced 1 | Number of Potentially Broken Antioxidant Fragments |
---|---|---|---|
Papain | Q9FRT8 | GFC | 0 |
Q43680 | EVY, VY, VYF, EAY, AY, AYV, VLK, KPS | 6 | |
Thermolysin | Q9FRT8 | GFC, FC, FCI, LK, LKK, NFC | 1 |
Q43680 | EVY, VY, VYF, EAY, AY, AYV, AYI, VYL, SIR, IR, IRN, AYL, VLK, LK, LKA, AFC, FC, EL, ELN, NKP, KP, KPS, NHL, HL, HLS | 9 |
Enzyme Scheme | Time (min) | ABTS Radical Scavenging Activity (Ascorbic Acid Equivalent, μM) | Iron Chelating Activity (EDTA Equivalent, μM) | ORAC (Trolox Equivalent, μM) |
---|---|---|---|---|
Thermolysin + MBA | 0 | 685.0 ± 333.8 | 3034.8 ± 216.4 | 127 ± 19 |
5 | 632.7± 288.6 | 5334.9 ± 45.3 | 125 ± 21 | |
10 | 683.2 ± 296.1 | 5337.8 ± 38.3 | 115 ± 17 | |
25 | 449.7 ± 301.4 | 5328.1 ± 17.4 | 98 ± 39 | |
45 | 511.0 ± 255.7 | 5329.2 ± 35.4 | 116 ± 17 | |
60 | 493.9 ± 297.7 | 5336.1 ± 26.2 | 127 ± 17 | |
240 | 646.2 ± 324.7 | 5161.7 ± 280.2 | 127 ± 17 | |
0 | 513.7 ± 230.4 | 3005.5 ± 248.0 | 122 ± 17 | |
15 | 533.5 ± 332.0 | 472.4 ± 227.4 | 126 ± 8 | |
Pepsin + MBA | 25 | 535.3 ± 328.0 | 529.4 ± 234.1 | 96 ± 20 |
45 | 540.8 ± 307.3 | 500.2 ± 173.3 | 99 ± 35 | |
60 | 477.7 ± 316.6 | 490.7 ± 300.4 | 122 ± 12 | |
90 | 638.1 ± 340.2 | 594.3 ± 108.6 | 107 ± 42 | |
120 | 495.7 ± 342.3 | 434.3 ± 230.1 | 101 ± 46 | |
0 | 487.6 ± 338.4 | 448.5 ± 240.1 | 145 ± 38 | |
15 | 450.2 ± 283.0 | 4924.6 ± 119.4 | 105 ± 5 | |
25 | 474.1 ± 245.5 | 4901.0 ± 95.0 | 97 ± 31 | |
Pepsin-Pancreatin + MBA | 45 | 559.7 ± 197.0 | 4886.9 ± 182.8 | 121 ± 17 |
60 | 456.9 ± 260.3 | 4884.4 ± 105.6 | 125 ± 16 | |
90 | 552.9 ± 256.4 | 4794.9 ± 16.5 | 124 ± 16 | |
120 | 581.3 ± 242.9 | 4843.5 ± 155.5 | 127 ± 15 |
Peptide Sequence | Molecular Mass (Da) | Hydrophobicity (Kcal/mol) | Isoelectric Point | Charge |
---|---|---|---|---|
MD | 264 | 10.87 | 2.95 | –1 |
QSA | 304 | 9.63 | 5.49 | 0 |
EW | 333 | 9.44 | 3.27 | –1 |
LGW | 374 | 5.71 | 5.69 | 0 |
KK | 274 | 13.5 | 10.57 | 2 |
SVP | 301 | 8.04 | 5.18 | 0 |
DVAF | 450 | 9.87 | 3.05 | –1 |
Peptide Sequence | Molecular Mass (Da) | Hydrophobicity (Kcal/mol) | Isoelectric Point | Charge |
---|---|---|---|---|
KK | 274 | 14.50 | 10.57 | 2 |
DM | 264 | 10.87 | 3.02 | –1 |
SY | 268 | 7.65 | 5.38 | 0 |
W | 204 | 5.81 | 5.64 | 0 |
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Kusumah, J.; Real Hernandez, L.M.; Gonzalez de Mejia, E. Antioxidant Potential of Mung Bean (Vigna radiata) Albumin Peptides Produced by Enzymatic Hydrolysis Analyzed by Biochemical and In Silico Methods. Foods 2020, 9, 1241. https://doi.org/10.3390/foods9091241
Kusumah J, Real Hernandez LM, Gonzalez de Mejia E. Antioxidant Potential of Mung Bean (Vigna radiata) Albumin Peptides Produced by Enzymatic Hydrolysis Analyzed by Biochemical and In Silico Methods. Foods. 2020; 9(9):1241. https://doi.org/10.3390/foods9091241
Chicago/Turabian StyleKusumah, Jennifer, Luis M. Real Hernandez, and Elvira Gonzalez de Mejia. 2020. "Antioxidant Potential of Mung Bean (Vigna radiata) Albumin Peptides Produced by Enzymatic Hydrolysis Analyzed by Biochemical and In Silico Methods" Foods 9, no. 9: 1241. https://doi.org/10.3390/foods9091241