Bioactive Peptides from Germinated Soybean with Anti-Diabetic Potential by Inhibition of Dipeptidyl Peptidase-IV, α-Amylase, and α-Glucosidase Enzymes
Abstract
:1. Introduction
2. Results and Discussion
2.1. In Vitro Anti-Diabetic Activity of Germinated Soybean Protein Hydrolyzed Under Simulated Gastrointestinal Conditions
2.2. Identification of Potential Anti-Diabetic Peptides
3. Materials and Methods
3.1. Materials
3.2. Preparation of Germinated Soybean Protein Isolate
3.3. Simulation of Gastrointestinal Digestion
3.4. Measurement of the Enzymatic Inhibitory Activity
3.4.1. DPP-IV Inhibitory Activity
3.4.2. α-Amylase Inhibitory Activity
3.4.3. α-Glucosidase Inhibitory
3.5. Separation of DPP-IV Inhibitory Peptides by Semi-Preparative RP-HPLC
3.6. Identification of Anti-Diabetic Peptides by RP-HPLC-MS/MS
3.7. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
6GSPD | 6-day germinated soybean protein digest |
DM | diabetes mellitus |
DPP-IV | dipeptidyl peptidase IV |
MGAM | maltase-glucoamylase |
RP-HPLC-MS/MS | RP-HPLC coupled to tandem mass spectrometry |
SI | sucrase-isomaltase |
T2DM TFA | type 2 diabetes mellitus trifluoroacetic acid |
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Enzymatic Inhibitory Activity (IC50, mg/mL) * | ||||
---|---|---|---|---|
Samples | DPP-IV | α-Amylase | α-Glucosidase | |
Maltase Activity | Sucrase Activity | |||
Positive control 1 | 0.003 ± 0.000 a | 0.16 ± 0.01 a | 0.07 ± 0.01 a | 0.03 ± 0.00 a |
6GSPD | 1.49 ± 0.14 c | 1.70 ± 0.18 b | 3.73 ± 0.06 c | 2.90 ± 0.07 d |
>10 kDa | 1.18 ± 0.15 b | 4.80 ± 0.87 c | >10.00 d | 5.27 ± 0.16 e |
5–10 kDa | 0.91 ± 0.17 b | >10.00 | 3.56 ± 0.14 c | 2.20 ± 0.40 c |
<5 kDa | 2.21 ± 0.15 d | 8.30 ± 1.65 d | 2.56 ± 0.03 b | 1.23 ± 0.19 b |
Peptide Fraction | Mass | Protein Fragment | Peptide Sequence | Net Charge c | Hydrophobicity (kcal/mol) c | DPP-IV Inhibitory Peptides a | Antioxidant Peptides a | ACE Inhibitory Peptides a |
---|---|---|---|---|---|---|---|---|
F1 b | 834.3 | β-conglycinin α, α’and β-chain f(270–276) | NNDDRDS | −2 | +22.79 | DR, ND, NN | --- | --- |
1013.4 | β-conglycinin α and α’-chain f(295–303) | VVNPDNNEN | −2 | +17.79 | DN, NE, NN, NP, VN, VV | NEN | VNP | |
949.4 | β-conglycinin α and β-chain f(324–332) | LSSTEAQQS | −1 | +13.95 | QQ, QS, TE | --- | EA, ST, TE | |
958.4 | β-conglycinin α, α’and β-chain f(530–537) | NAENNQRN | 0 | +18.01 | AE, NA, NN, NQ, RN | --- | --- | |
777.4 | β-conglycinin α’-chain f(604–610) | IKSQSES | 0 | +15.36 | ES, KS, QS | --- | --- | |
981.4 | Glycinin G1 subunit f(112–119) | EEPQQPQQ | −2 | +18.52 | EP, QP, PQ, QQ | --- | PQ | |
874.4 | Glycinin G1 subunit f(121–128) | GQSSRPQD | 0 | +17.10 | GQ, PQ, QD, QS, RP | --- | GQ, PQ, RP | |
887.4 | Glycinin G2 and G1 subunit f(181–188) | LAGNQEQE | −2 | +17.95 | AG, LA, NQ, QE | --- | AG, LA | |
787.4 | Glycinin G1 subunit f(465–471) | NLKSQQA | +1 | +12.80 | KS, NL, QA, QQ | LK | --- | |
972.4 | Glycinin G2 subunit f(109–116) | QEPQESQQ | −2 | +18.84 | EP, ES, PQ, QE, QQ | --- | PQ | |
1150.6 | Glycinin G2 subunit f(120–128) | SQRPQDRHQ | +1 | +20.40 | DR, PQ, QD, RH, RP | RHQ | PQ, RP | |
1202.5 | Glycinin G2 subunit f(193–203) | QQQQQGGSQSQ | 0 | +16.50 | GG, QG, QQ, QS | --- | GG, GS, QG | |
1387.7 | Glycinin G2 subunit f(193–205) | QQQQQGGSQSQKG | +1 | +20.46 | GG, KG, QG, QQ, QS | --- | GG, GS, KG, QG, QK | |
1244.6 | Glycinin G2 and G5 subunit f(198–207) | PETMQQQQQQ | −1 | +15.87 | ET, MQ, QQ, TM | --- | --- | |
1311.5 | P34 Probable thiol protease f(250–261) | SDESTESETEQA | −5 | +29.21 | ES, ET, QA, TE | --- | ST, TE | |
F2 | 1546.7 | Glycinin G2 subunit f(238–251) | RNLQGENEEEDSGA | −4 | +32.34 | GA, GE, NE, NL, QG, RN | --- | GA, GE, LQ, QG, SG |
843.5 | Glycinin G4 subunit f(451–458) | VTRGQGKV | +2 | +14.89 | GQM KVM QG, RG, TR, VT | --- | GK, GQ, QG, RG, GKV, VTR | |
644.4 | P34 Probable thiol protease f(143–148) | KKGVIT | +2 | +13.32 | GV, KG, KK, VI | --- | GV, KG | |
1555.6 | P34 Probable thiol protease f(248–261) | IMSDESTESETEQA | −5 | +27.42 | ES, ET, IM, QA, TE | --- | ST, TE | |
F3 | 1497.7 | Glycinin G1 subunit f(37–50) | NALKPDNRIESEGG | −1 | +26.14 | AL, DN, EG, ES, GG, KP, NA, NR, RI | LK, KP, LKP | EG, GG, IE, KP, LKP |
1433.7 | Glycinin G1 subunit f(329–342) | SSPDIYNPQAGSVT | −1 | +14.43 | AG, NP, PQ, QA, SP, SV, VT, YN | IY | AG, GS, IY, PQ, YN, AGS | |
1497.7 | Glycinin G2 subunit f(34–47) | NALKPDNRIESEGG | −1 | +26.14 | AL, DN, EG, ES, GG, KP, NA, NR, RI | LK, KP, LKP | EG, GG, IE, KP, LKP | |
1956.9 | Glycinin G2 subunit f(312–329) | RQNIGQNSSPDIYNPQAG | 0 | +19.57 | AG, GQ, PQ, QA, QN, SP, YN | IY | AG, GQ, IG, IY, PQ, YN | |
1497.7 | Glycinin G3 subunit f(37–50) | NALKPDNRIESEGG | −1 | +26.14 | AL, DN, EG, ES, GG, KP, NA, NR, RI | LK, KP, LKP | EG, GG, IE, KP, LKP | |
1261.6 | Glycinin G4 subunit f(486–497) | VVAEQAGEQGFE | −3 | +21.00 | AE, AG, GE, GF, QA, QG, VA, VV | --- | AG, GE, GF, QG | |
882.5 | β-conglycinin α-chain f(190–196)β-conglycinin α’-chain f(206–212) | HKNKNPF | +2 | +15.96 | NP, PF | --- | HK, NK |
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González-Montoya, M.; Hernández-Ledesma, B.; Mora-Escobedo, R.; Martínez-Villaluenga, C. Bioactive Peptides from Germinated Soybean with Anti-Diabetic Potential by Inhibition of Dipeptidyl Peptidase-IV, α-Amylase, and α-Glucosidase Enzymes. Int. J. Mol. Sci. 2018, 19, 2883. https://doi.org/10.3390/ijms19102883
González-Montoya M, Hernández-Ledesma B, Mora-Escobedo R, Martínez-Villaluenga C. Bioactive Peptides from Germinated Soybean with Anti-Diabetic Potential by Inhibition of Dipeptidyl Peptidase-IV, α-Amylase, and α-Glucosidase Enzymes. International Journal of Molecular Sciences. 2018; 19(10):2883. https://doi.org/10.3390/ijms19102883
Chicago/Turabian StyleGonzález-Montoya, Marcela, Blanca Hernández-Ledesma, Rosalva Mora-Escobedo, and Cristina Martínez-Villaluenga. 2018. "Bioactive Peptides from Germinated Soybean with Anti-Diabetic Potential by Inhibition of Dipeptidyl Peptidase-IV, α-Amylase, and α-Glucosidase Enzymes" International Journal of Molecular Sciences 19, no. 10: 2883. https://doi.org/10.3390/ijms19102883