Purification and Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides and the Antihypertensive Effect of Chlorella sorokiniana Protein Hydrolysates
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation of Hot Water Extract and Hydrolysate of C. sorokiniana
2.3. Chemical Analyses
2.4. Measurement of Peptide Content
2.5. In Vitro Assay for ACE-Inhibitory Activity
2.6. In Vitro Gastrointestinal Digestion
2.7. Size Exclusion Chromatography
2.8. Purification of ACE-Inhibitory Peptide
2.9. Sequence Analysis
2.10. Animals and In Vivo Measurement of Blood Pressure
2.11. Statistical Analysis
3. Results and Discussion
3.1. Soluble Protein Content, Peptide Content, Yield, and IC50
3.2. In Vitro Stability of C. sorokiniana–Derived ACE-Inhibitory Peptides
3.3. Isolation and Purification of ACE-Inhibitory Peptide
3.4. Amino Acid Sequences and ACE-Inhibitory Activity
3.5. Antihypertensive Effect of C. sorokiniana Protein Hydrolysate
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample | Soluble Protein (mg/g) | Peptide Content (mg/g) | Yield 1 | IC50 2 (mg/mL) |
---|---|---|---|---|
HWE 3 | 379.9 ± 1.5 | 179.7 ± 2.1 | 4.0 | 1.070 ± 0.020 |
PX-1 4 | 482.0 ± 2.2 | 260.4 ± 2.0 | 19.1 | 0.043 ± 0.001 |
PX-2 5 | 566.6 ± 3.5 | 298.6 ± 2.0 | 22.7 | 0.043 ± 0.002 |
PN-1 6 | 574.8 ± 2.3 | 332.8 ± 3.0 | 28.1 | 0.035 ± 0.002 |
PN-2 7 | 610.6 ± 3.8 | 341.6 ± 3.1 | 31.2 | 0.042 ± 0.001 |
Protease | IC50 (mg Peptide/mL) |
---|---|
Control | 0.035 ± 0.002 |
Pepsin 1 | 0.044 ± 0.001 |
Pepsin + Pancreatin 2 | 0.044 ± 0.001 |
Fraction | Molecular Weight (Da) | Inhibition (%) | Peptide Content (mg/mL) | IER 1 (%/mg/mL) | IC50 (mg/mL) |
---|---|---|---|---|---|
A | 1400–1180 | 74.0 | 0.210 | 350 | — 2 |
B | 1180–910 | 30.6 | 0.027 | 1130 | 0.0450 |
C | 910–740 | 58.2 | 0.025 | 2230 | 0.0187 |
D | 680–590 | 48.0 | 0.064 | 750 | — |
E | 460–370 | 73.2 | 0.057 | 1280 | 0.0160 |
F | 340–270 | 68.6 | 0.034 | 2020 | 0.0150 |
G | 200–250 | 40.0 | 0.053 | 760 | — |
Peak | ACE Inhibitory (%) | Peptide Content (mg/mL) | IER (%/mg/mL) |
---|---|---|---|
F1 | 14.3 | 0.01 | 1430 |
F2 | 19.1 | 0.01 | 1910 |
F3 | 41.2 | 0.02 | 2060 |
F4 | 36.8 | 0.01 | 3680 |
F5 | 72.4 | 0.02 | 3621 |
F6 | 75.7 | 0.03 | 2523 |
F7 | 54.3 | 0.01 | 5425 |
F8 | 86.1 | 0.01 | 8613 |
F9 | 95.1 | 0.01 | 9510 |
F10 | 87.7 | 0.01 | 8770 |
Peak | Sequence | IC50 (μM) |
---|---|---|
F7 | Trp–Val | 307.61 ± 0.01 |
F8 | Val–Trp | 0.58 ± 0.02 |
F9 | Ile–Trp | 0.50 ± 0.01 |
F10 | Leu–Trp | 1.11 ± 0.02 |
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Lin, Y.-H.; Chen, G.-W.; Yeh, C.H.; Song, H.; Tsai, J.-S. Purification and Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides and the Antihypertensive Effect of Chlorella sorokiniana Protein Hydrolysates. Nutrients 2018, 10, 1397. https://doi.org/10.3390/nu10101397
Lin Y-H, Chen G-W, Yeh CH, Song H, Tsai J-S. Purification and Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides and the Antihypertensive Effect of Chlorella sorokiniana Protein Hydrolysates. Nutrients. 2018; 10(10):1397. https://doi.org/10.3390/nu10101397
Chicago/Turabian StyleLin, Yu-Hsin, Guan-Wen Chen, Chin Hsi Yeh, Helena Song, and Jenn-Shou Tsai. 2018. "Purification and Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides and the Antihypertensive Effect of Chlorella sorokiniana Protein Hydrolysates" Nutrients 10, no. 10: 1397. https://doi.org/10.3390/nu10101397