Deer Skin Collagen Peptides Bound to Calcium: In Vitro Gastrointestinal Simulation of Digestion, Cellular Uptake and Analysis of Antioxidant Activity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Preparation of Ca-DSCP
2.2.1. Single-Factor Experiments
2.2.2. Response Surface Experiments
2.2.3. Determination of Calcium Chelation Rate
2.3. Amino Acid Analysis
2.4. Zeta Potential and XRD of DSCP and Ca-DSCP
2.5. UV–vis and FTIR of DSCP and Ca-DSCP
2.6. Stability of Ca-DSCP
2.6.1. Acid–Base Stability and Thermal Stability
2.6.2. In Vitro Gastrointestinal Digestion
2.7. Cellular Experiments
2.7.1. Caco-2 Cell Culture
2.7.2. Cytotoxicity Test of Caco-2 Cells
2.7.3. Caco-2 Cells Monolayer Calcium Transport Studies
2.7.4. Measurement of Oxidative Stress Indicators
2.8. Antioxidant Capacity of DSCP and Ca-DSCP
2.8.1. DPPH Radical Scavenging Ability Assay
2.8.2. Hydroxyl Radical Scavenging Ability Assay
2.9. Bioinformatics and Molecular Docking
2.10. Statistical Analysis
3. Results and Discussion
3.1. Effect of Different Process Conditions on the Preparation of Ca-DSCP
3.2. Response Surface Analysis for the Preparation of Ca-DSCP
3.3. Amino Acid Composition
3.4. Zeta Potential and XRD Analysis
3.5. Structural Characterization of UV–vis and FTIR Analysis
3.6. Acid–Base Stability and Thermal Stability Analysis
3.7. In Vitro Gastrointestinal Digestion
3.8. Caco-2 Cytotoxicity Test
3.9. Caco-2 Cells Monolayer Calcium Transport Studies
3.10. Analysis of Oxidative Stress Indicators
3.11. DPPH Radical Scavenging Capacity of DSCP and Ca-DSCP
3.12. Hydroxyl Radical Scavenging Capacity of DSCP and Ca-DSCP
3.13. Bioinformatics and Molecular Docking
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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No. | A-Mass Ratio of Peptide to Calcium | B-pH | C-Temperature | Chelation Rate % |
---|---|---|---|---|
1 | 5:1 | 9 | 50 | 61.52 |
2 | 3:1 | 9 | 65 | 46.48 |
3 | 7:1 | 9 | 65 | 50.83 |
4 | 7:1 | 8 | 50 | 54.58 |
5 | 5:1 | 9 | 50 | 59.77 |
6 | 7:1 | 10 | 50 | 53.75 |
7 | 5:1 | 9 | 50 | 61.19 |
8 | 7:1 | 9 | 35 | 52.74 |
9 | 3:1 | 10 | 50 | 51.27 |
10 | 5:1 | 10 | 65 | 52.27 |
11 | 5:1 | 8 | 65 | 52.99 |
12 | 3:1 | 8 | 50 | 51.30 |
13 | 5:1 | 8 | 35 | 55.30 |
14 | 5:1 | 10 | 35 | 55.51 |
15 | 5:1 | 9 | 50 | 59.11 |
16 | 3:1 | 9 | 35 | 49.57 |
17 | 5:1 | 9 | 50 | 60.16 |
Source | Sum of Squares | df | Mean Square | F-Value | p-Value | Significant |
---|---|---|---|---|---|---|
Model | 305.62 | 9 | 33.96 | 53.59 | <0.0001 | ** |
A-Mass ratio of peptide to calcium | 21.95 | 1 | 21.95 | 34.63 | 0.0006 | ** |
B-pH | 0.2244 | 1 | 0.2244 | 0.3542 | 0.5705 | |
C-Temperature | 13.97 | 1 | 13.97 | 22.04 | 0.0022 | * |
AB | 0.1681 | 1 | 0.1681 | 0.2653 | 0.6224 | |
AC | 0.3782 | 1 | 0.3782 | 0.5968 | 0.4651 | |
BC | 0.2209 | 1 | 0.2209 | 0.3486 | 0.5735 | |
A2 | 144.90 | 1 | 144.90 | 228.64 | <0.0001 | ** |
B2 | 12.95 | 1 | 12.95 | 20.43 | 0.0027 | * |
C2 | 88.18 | 1 | 88.18 | 139.14 | <0.0001 | ** |
Residual | 4.44 | 7 | 0.6337 | |||
Lack of fit | 0.4515 | 3 | 0.1505 | 0.1511 | 0.9238 | not significant |
Pure error | 3.98 | 4 | 0.9962 | |||
Cor total | 310.06 | 16 |
Peptide | Mass (Da) | Toxicity | Hydrophobicity (Kcal/mol) | Post-Digestive Fragments | Binding Energy (kcal/mol) | Hydrogen Bonds | Structural Formula | |
---|---|---|---|---|---|---|---|---|
Quantity | Binding Sites | |||||||
A | 1014.11 | None | 16.14 | PGPGPGPGPGPGA | −12.1 | 13 | VAL561, VAL604, VAL606, ILE559, GLY367, GLY419, THR,560, THR609, ARG326 | |
B | 1097.2 | None | 16.93 | PGPGPGPGPGPGPG | −11.9 | 18 | LEU365, LEU468, ASN469, ASN517, VAL369, VAL420, VAL463, VAL465, VAL512, VAL608, GLY419, ILE416, ILE559, ASP442, ALA510 | |
C | 1044.14 | None | 15.72 | PAAGGPFPGHH | −11.2 | 17 | ILE416, ILE559, GLY367, GLY423, GLY558, VAL467, VAL561, VAL606, THR560, ASN469, ASN517, ASP422, ARG362 | |
D | 1154.29 | None | 17.77 | PPGEPGPPGPRP | −11.0 | 14 | VAL420, VAL467, VAL512, VAL516, VAL604, LEU365, LEU557, ALA510, ILE559, THR560, HIS516, | |
E | 1142.24 | None | 16.91 | PGPGPGPGQAPPGG | −10.9 | 6 | ILE559, VAL420, VAL606, GLY367, ARG326 | |
F | 1003.08 | None | 14.26 | PGPSPGPGPSPG | −10.9 | 13 | VAL418, VAL420, VAL463, VAL465, VAL467, ASP422, THR560, ARG326 | |
G | 873.96 | None | 14.2 | PGPAGPPGAPG | −10.9 | 8 | VAL369, VAL418, VAL420, VAL463, VAL465, ASP422, ASN469, IEU365 | |
H | 1171.34 | None | 15.11 | MPGPGPGPGPGPGP | −10.8 | 15 | ARG326, GLN563, GLY367, GLY419, GLY564, ASN469, VAL512, ILE559, LEU557, ALA510, VAL606 |
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Du, R.; Sun, L.; Liu, J.; Gao, F.; Guo, X.; Shi, M.; Guo, P.; Chen, W.; Zong, Y.; Geng, J.; et al. Deer Skin Collagen Peptides Bound to Calcium: In Vitro Gastrointestinal Simulation of Digestion, Cellular Uptake and Analysis of Antioxidant Activity. Nutrients 2024, 16, 2585. https://doi.org/10.3390/nu16162585
Du R, Sun L, Liu J, Gao F, Guo X, Shi M, Guo P, Chen W, Zong Y, Geng J, et al. Deer Skin Collagen Peptides Bound to Calcium: In Vitro Gastrointestinal Simulation of Digestion, Cellular Uptake and Analysis of Antioxidant Activity. Nutrients. 2024; 16(16):2585. https://doi.org/10.3390/nu16162585
Chicago/Turabian StyleDu, Rui, Li Sun, Jinze Liu, Fusheng Gao, Xiangjuan Guo, Meiling Shi, Pengli Guo, Weijia Chen, Ying Zong, Jianan Geng, and et al. 2024. "Deer Skin Collagen Peptides Bound to Calcium: In Vitro Gastrointestinal Simulation of Digestion, Cellular Uptake and Analysis of Antioxidant Activity" Nutrients 16, no. 16: 2585. https://doi.org/10.3390/nu16162585