Degree of Hydrolysis Regulated by Enzyme Mediation of Wheat Gluten Fibrillation: Structural Characterization and Analysis of the Mechanism of Action
Abstract
:1. Introduction
2. Results
2.1. Optimal Enzyme–Substrate Ratio
2.2. Analysis SDS-PAGE
2.3. ThT Fluorescence Intensity
2.4. Analysis FTIR Absorption Spectra
2.5. Analysis of Surface Hydrophobicity and ζ-Potential
2.6. Analysis of SEM
2.7. Analysis of TEM
3. Discussion
3.1. Possible Mechanism Analysis of AFs’ Formation Process of WG
3.2. Concluding Remarks and Future Perspectives
4. Materials and Methods
4.1. Materials
4.2. Hydrolysis of WG
4.3. Preparation of AFs
4.4. SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE)
4.5. ThT Fluorescence Assay
4.6. Surface Hydrophobicity Analysis
4.7. ζ-Potential Measurements
4.8. FTIR Spectroscopy
4.9. SEM
4.10. TEM
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Degrees of Hydrolysis | Enzyme–Substrate Ratio | ||
---|---|---|---|
1:100 | 1:500 | 1:1000 | |
DH4 | 1.3 ± 0.1 (h) a | 2.2 ± 0.2 (h) a | 4.3 ± 0.2 (h) b |
DH5 | 2.7 ± 0.15 (h) a | 3.5 ± 0.15 (h) a | 5.8 ± 0.32 (h) b |
DH6 | 4.5 ± 0.25 (h) a | 4.5 ± 0.18 (h) a | 7 ± 0.31 (h) b |
DH7 | 6.1 ± 0.25 (h) a | 7.5 ± 0.25 (h) a | 10 ± 0.25 (h) b |
DH8 | 7.6 ± 0.15 (h) a | 8.5 ± 0.15 (h) a | 12 ± 0.31 (h) b |
Degrees of Hydrolysis | Area (%) | ||||
---|---|---|---|---|---|
β-Sheet | Random Coil | α-Helix | Antiparallel β-Sheet | β-Turn | |
DH4 | 30.7484 | 17.3761 | 34.5704 | 9.0298 | 12.2753 |
DH5 | 31.9271 | 15.7266 | 33.3201 | 8.067 | 10.9592 |
DH6 | 31.3268 | 14.8518 | 34.9059 | 8.1665 | 10.7489 |
DH7 | 30.1374 | 16.1288 | 34.1976 | 8.2575 | 11.2787 |
DH8 | 29.7842 | 17.635 | 31.6121 | 8.8613 | 12.1073 |
Degrees of Hydrolysis | Area (%) | |||||
---|---|---|---|---|---|---|
Δβ-Sheet * | Β-Sheet | Random Coil | α-Helix | Antiparallel β-Sheet | β-Turn | |
DH4 | 13.037 | 43.7854 | 25.1712 | 14.3445 | 5.6842 | 10.6739 |
DH5 | 14.3558 | 46.2829 | 16.7061 | 17.7164 | 6.0611 | 12.9063 |
DH6 | 19.1468 | 50.4736 | 21.5308 | 12.6679 | 5.7047 | 9.48678 |
DH7 | 14.9632 | 45.1006 | 28.0488 | 10.3465 | 5.7071 | 10.5248 |
DH8 | 14.6054 | 44.3896 | 15.5662 | 23.7324 | 6.1335 | 10.1332 |
Amide I Structural Assignment | Wavenumber (cm−1) |
---|---|
cross-β | 1611–1637 |
Random coil | 1637–1647 |
α-helix | 1647–1662 |
β-turn | 1662–1678; 1689–1699 |
antiparallel β-sheet | 1679–1688 |
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Zhang, H.; Lv, S.; Ren, F.; Liu, J.; Wang, J. Degree of Hydrolysis Regulated by Enzyme Mediation of Wheat Gluten Fibrillation: Structural Characterization and Analysis of the Mechanism of Action. Int. J. Mol. Sci. 2023, 24, 13529. https://doi.org/10.3390/ijms241713529
Zhang H, Lv S, Ren F, Liu J, Wang J. Degree of Hydrolysis Regulated by Enzyme Mediation of Wheat Gluten Fibrillation: Structural Characterization and Analysis of the Mechanism of Action. International Journal of Molecular Sciences. 2023; 24(17):13529. https://doi.org/10.3390/ijms241713529
Chicago/Turabian StyleZhang, Huijuan, Shihao Lv, Feiyue Ren, Jie Liu, and Jing Wang. 2023. "Degree of Hydrolysis Regulated by Enzyme Mediation of Wheat Gluten Fibrillation: Structural Characterization and Analysis of the Mechanism of Action" International Journal of Molecular Sciences 24, no. 17: 13529. https://doi.org/10.3390/ijms241713529