Novel Hybrid Peptide Cecropin A (1–8)-LL37 (17–30) with Potential Antibacterial Activity
Abstract
:1. Introduction
2. Results
2.1. Primary Structural Parameters of Peptides
2.2. Antibacterial and Hemolytic Activity
2.3. Sterilization Speed and Efficacy
2.4. Disruption of Bacterial Cell Surface Structure Integrity
2.5. Synergistic Assay
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Antibiotics
4.2. Design, Biological Information Analysis, and Synthesis of Hybrid Peptides
4.3. Determination of Minimal Inhibitory Concentrations (MICs)
4.4. Hemolytic Assay
4.5. Antibacterial Studies
4.6. SEM Analysis of Bacterial Cells
4.7. Synergy Assays
4.8. Statistical Analyses
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Peptide | Amino Acid Sequence | Theoretical MW | Measured MW |
---|---|---|---|
C-L | KWKLFKKIFKRIVQRIKDFLRN | 2905.70 | 2905.60 |
L | LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES | 4490.57 | 4490.60 |
C | KWKLFKKIEKVGQNIRDGIIKAGPAVAVVGQATQIAK | 4010.60 | 4010.82 |
Peptide | Structure | Net Charge | Helical Residue (%) | Hydrophobic Residue (%) |
---|---|---|---|---|
C-L | α-helical | +8 | 90.9 | 45 |
L | α-helical | +6 | 81.1 | 35 |
C | α-helical | +7 | 81.1 | 45 |
Bacteria Strains | Antimicrobial Peptide | p-Value | |||
---|---|---|---|---|---|
C | L | C-L | |||
MIC (µg/mL) | E. coli K88 | 64 ± 0.72 a | 61 ± 1.78 b | 8 ± 0.33 c | <0.01 |
E. coli CVCC 245 | 4 ± 0.28 c | 84 ± 1.54 a | 7 ± 0.29 b | <0.01 | |
S. aureus CVCC 26003 | 128 ± 7.57 a | 36 ± 1.55 b | 7 ± 0.28 c | <0.01 | |
S. aureus ATCC 25923 | 198 ± 10.1 a | 58 ± 1.68 b | 2 ± 0.03 c | <0.01 | |
L. mono. CVCC 1599 | 64 ± 2.99 a | 13 ± 0.43 b | 2 ± 0.06 c | <0.01 | |
M. luteus CVCC 28001 | 154 ± 5.49 a | 90 ± 1.43 b | 16 ± 0.18 c | <0.01 | |
HC50 (µg/mL) | Sheep erythrocyte cell | 169 ± 15.1 b | 32 ± 0.68 c | 221 ± 3.27 a | <0.01 |
Bacteria Strains | Time (h) | Antimicrobial Peptides | p-Value | ||
---|---|---|---|---|---|
C | L | C-L | |||
Viable S. aureus concentration (×108 CFU/mL) | 1 | 0.54 ± 0.007 b | 0.90 ± 0.020 a | 0.43 ± 0.017 c | <0.01 |
2 | 0.40 ± 0.035 b | 0.53 ± 0.026 a | 0.30 ± 0.012 c | <0.01 | |
3 | 0.21 ± 0.014 a | 0.23 ± 0.011 a | 0.18 ± 0.020 b | 0.014 | |
4 | 0.09 ± 0.005 | 0.09 ± 0.002 | 0.08 ± 0.003 | 0.079 | |
5 | 0.01 ± 0.0005 | 0.01 ± 0.0003 | 0.01 ± 0.0001 | 0.298 | |
Viable E. coli concentration (×108 CFU/mL) | 1 | 0.44 ± 0.028 a | 0.44 ± 0.027 a | 0.29 ± 0.015 b | <0.01 |
2 | 0.30 ± 0.021 b | 0.35 ± 0.014 a | 0.16 ± 0.026 c | <0.01 | |
3 | 0.15 ± 0.014 a | 0.14 ± 0.020 a | 0.05 ± 0.003 b | <0.01 | |
4 | 0.06 ± 0.002 | 0.05 ± 0.002 | 0.05 ± 0.003 | 0.061 | |
5 | 0.01 ± 0.0002 | 0.01 ± 0.003 | 0.01 ± 0.0001 | 0.659 |
Combination Group | Fractional Inhibitory Concentration (FIC) Index | p-Value | ||||
---|---|---|---|---|---|---|
C-L + Chloramphenicol | C-L + Thiamphenicol | C-L + Neomycin Sulfate | C-L + Penicillin G | C-L + Kanamycin | ||
E. coli CVCC 245 | 1.00 ± 0.141 c | 1.50 ± 0.001 a | 0.313 ± 0.001 d | 1.50 ± 0.003 a | 1.25 ± 0.002 b | <0.01 |
S. aureus ATCC 25923 | 0.375 ± 0.005 c | 0.188 ± 0.002 e | 0.250 ± 0.004 d | 1.13 ± 0.005 a | 0.750 ± 0.003 b | <0.01 |
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Wei, X.-B.; Wu, R.-J.; Si, D.-Y.; Liao, X.-D.; Zhang, L.-L.; Zhang, R.-J. Novel Hybrid Peptide Cecropin A (1–8)-LL37 (17–30) with Potential Antibacterial Activity. Int. J. Mol. Sci. 2016, 17, 983. https://doi.org/10.3390/ijms17070983
Wei X-B, Wu R-J, Si D-Y, Liao X-D, Zhang L-L, Zhang R-J. Novel Hybrid Peptide Cecropin A (1–8)-LL37 (17–30) with Potential Antibacterial Activity. International Journal of Molecular Sciences. 2016; 17(7):983. https://doi.org/10.3390/ijms17070983
Chicago/Turabian StyleWei, Xu-Biao, Ru-Juan Wu, Da-Yong Si, Xiu-Dong Liao, Lu-Lu Zhang, and Ri-Jun Zhang. 2016. "Novel Hybrid Peptide Cecropin A (1–8)-LL37 (17–30) with Potential Antibacterial Activity" International Journal of Molecular Sciences 17, no. 7: 983. https://doi.org/10.3390/ijms17070983