Study on the Inhibitory Effect of Bioactive Peptides Derived from Yak Milk Cheese on Cholesterol Esterase
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Instruments and Equipment
2.3. Experimental Methods
2.3.1. Prediction of Physicochemical Properties of Peptides RK7, KQ7, QP13, VN10, TL9, LQ10, and SN12
2.3.2. Structure Optimization and Processing of Peptides and Cholesterol Esterase
2.3.3. Molecular Docking
2.3.4. GROMACS Molecular Dynamics Simulation
2.3.5. Synthesis and CE Inhibitory Activity Assay of Peptides RK7, KQ7, QP13, and VN10
2.3.6. Simulating the Impact of Gastrointestinal Digestion on α-Amylase Inhibitory Activity
2.3.7. Data Processing
3. Results and Discussion
3.1. Physicochemical Characterisation of RK7, KQ7 QP13, TL9, VN10, SN12, and LQ10 Peptides
3.2. Molecular Docking Using LibDock
3.2.1. Reliability Verification of Molecular Docking Methods
3.2.2. Molecular Docking of CE Inhibitory Peptides
3.3. GROMACS Analysis of the Molecular Dynamics Stability of Peptide–CE Complexes
3.3.1. RMSD, RMSF, Hydrogen Bonds, SASA, and Gibbs Free Energy Stability Analysis
3.3.2. MM/GBSA Calculations and Analyses
3.3.3. Analysis of Interactions in the 0–100 ns Process of Molecular Dynamics
3.4. Synthesis and Validation of CE Inhibitory Activity of Peptides RK7, KQ7, QP13 and VN10
3.4.1. Synthesis of Peptides RK7, KQ7, QP13 and VN10
3.4.2. Validation of In Vitro Activity of Peptides RK7, KQ7, QP13, and VN10
3.4.3. Simulating the Effect of Gastrointestinal Digestion on CE Inhibition Activity
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Peptide Sequence | Molecular Weight/(Da) | Isoelectric Point | Net Charge | Hydrophobic Amino Acids | Proportion of Hydrophobic Amino Acids |
---|---|---|---|---|---|
RK7 | 874.90 | 11.57 | 3 | P, I | 42.86% |
KQ7 | 779.50 | 9.84 | 1 | V, L, P | 71.42% |
QP13 | 1392.60 | 6.58 | 0 | P, V, L, G, F | 61.54% |
TL9 | 968.19 | 3.32 | 0 | P, V, F, L | 88.89% |
VN10 | 1100.26 | 3.63 | 0 | V, P, F, G, I | 70.00% |
LQ10 | 1300.50 | 4.34 | 0 | L, P, V, M, F | 66.67% |
SN12 | 1126.37 | 3.70 | 0 | L, V, P, F, G, I | 80.00% |
RK7 | KQ7 | QP13 | VN10 | TL9 | LQ10 | SN12 | |
---|---|---|---|---|---|---|---|
Hydrogen bonds | Thr68 Ala67 Gln66 Asn118 Ala117 Asn122 Arg423 Pro425 Asn122 | His435 Asp437 Arg423 Met424 Asn118 Ser422 | Asp437 Arg423 Ser422 Pro425 Asn118 | Arg423 Try75 Gln71 Pro425 Met424 | Try75 Arg423 Ile426 | Arg423 Tyr75 Pro425 Tyr75 | Asn118 Tyr75 Asn118 |
Hydrophobic interactions | Met424 Leu69 Phe119 | Arg423 Met424 Ala117 Phe119 | Ile426 Phe119Ala117 Leu69 Leu124 Ala117 | Ile426 Pro425Met424 Arg423 Leu69 Tyr75 | Leu69 Tyr75 Pro425Met424 Ile426 Pro425 Ile426 Ala117 | Leu69 Tyr75 Ile426 | Phe119 Ala117 Leu69Met111 Tyr75 |
Van der Waals forces | Ile426 Lys445 Tyr123 Leu124 Met424 Pro425 Tyr75 Leu69 Met111 Gly112 | Tyr427 Leu323 Gln440 Pro425 Leu120 Leu69 Gln71 Asn122 Ala436 | Asp434 Lys445 Asn118 Met424 Ile426 Tyr75 Thr68 Tyr427 Tyr123 Leu69 Gln440 Asn112 Phe119 Arg63 | Gln440 Ser422 Asp434 His435 Ala436 Thr68 Leu323 Tyr427 | Gln71 Thr68 Met111 Ala67 Arg63 Ser422 Asp437 Tyr427 Asn118 Phe119 Thr68 | Tyr453 Pro425 Gly452 Met424 Thr68 Gln71 Ala117 Tyr427 | Leu274 Leu120 Ala113 Gly116 Gly112 Asp72 Thr74 Ala67 Tyr75 Arg63 Gln71 Thr70 |
Electrostatic interactions | — | — | Asp72 | Met424Asp437 | Arg423 | — | — |
(kcal/mol) | RK7 | KQ7 | QP13 | VN10 |
---|---|---|---|---|
VDWAALS | −89.74 ± 0.01 | −74.74 ± 1.38 | −84.13 ± 2.79 | −68.20 ± 0.15 |
ΔEEL | −25.73 ± 3.86 | −72.67 ± 3.27 | −50.58 ± 7.24 | −152.73 ± 1.87 |
ΔEGB | 65.11 ± 1.11 | 91.60 ± 3.06 | 106.08 ± 7.32 | 229.91 ± 7.28 |
ΔEsurf | −10.99 ± 0.01 | −9.44 ± 0.20 | −11.34 ± 0.02 | −10.06 ± 0.11 |
ΔGgas | −115.46 ± 3.86 | −147.40 ± 3.55 | −134.71 ± 7.75 | −220.94 ± 1.87 |
ΔGsolv | 54.12 ± 1.11 | 82.16 ± 3.07 | 94.74 ± 7.32 | 219.84 ± 7.28 |
ΔTotal | −61.34 ± 4.02 | −65.24 ± 4.69 | −39.97 ± 10.66 | −1.09 ± 7.52 |
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Wang, P.; Song, X.; Liang, Q. Study on the Inhibitory Effect of Bioactive Peptides Derived from Yak Milk Cheese on Cholesterol Esterase. Foods 2024, 13, 2970. https://doi.org/10.3390/foods13182970
Wang P, Song X, Liang Q. Study on the Inhibitory Effect of Bioactive Peptides Derived from Yak Milk Cheese on Cholesterol Esterase. Foods. 2024; 13(18):2970. https://doi.org/10.3390/foods13182970
Chicago/Turabian StyleWang, Peng, Xuemei Song, and Qi Liang. 2024. "Study on the Inhibitory Effect of Bioactive Peptides Derived from Yak Milk Cheese on Cholesterol Esterase" Foods 13, no. 18: 2970. https://doi.org/10.3390/foods13182970