Bacterial Cellulose Containing Combinations of Antimicrobial Peptides with Various QQ Enzymes as a Prototype of an “Enhanced Antibacterial” Dressing: In Silico and In Vitro Data
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Antibacterial Combinations Based on Antibacterial Agents and Enzymes Loaded onto Bacterial Cellulose (BC)
2.3. Water Absorption and Scanning Electron Microscopy (SEM) Analysis
2.4. Computational Methods
3. Results
3.1. Enzymes for Hydrolysis of G(−) Bacteria Signal Molecules
3.1.1. Physical–Chemical Interactions in Simulated Models of Enzyme–AMP Combinations
3.1.2. Antimicrobial Activity of AMP Combinations with AHL-Hydrolyzing Enzymes
3.2. Enzymes for Hydrolysis of G(+) Bacteria Signal Molecules in Combination with AMPs
3.2.1. Physical–Chemical Interactions in Simulated Models of AM Combinations with Peptidases
3.2.2. Antimicrobial Activity of BC Samples Loaded with AMPs in Combination with Peptidases
3.3. Absorption Capacity of BC Samples
3.4. Scanning Electron Microscopy of the Surface of BC Samples
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AHL | N-acyl homoserine lactone |
AIP | Autoinducing peptide |
AMP | Antimicrobial peptide |
BC | Bacterial cellulose |
CFU | Colony-forming unit |
His6-OPH | Hexahistidine-tagged organophosphorus hydrolase |
Quorum quenching | |
PON2 | Paraoxonase 2 |
References
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Antimicrobial Agent | Enzyme | Affinity, kJ × mol−1 | Occupied Area, % | ||
---|---|---|---|---|---|
Mean | Median | Total | Near Active Sites | ||
Dermicidin | Acylase | −20.4 | −20.1 ± 0.9 | 13.5 | 0.6 |
Lactonase | −14.6 | −15.1 ± 1.0 | 30.7 | 0.7 | |
PON2 | −14.7 | −14.6 ± 0.5 | 16.8 | 0 | |
His6-OPH | −15.2 | −15.1 ± 0.9 | 10.0 | 0.1 | |
Polymyxin B | Acylase | −25.3 | −24.7 ± 1.5 | 3.0 | 0 |
Lactonase | −28.6 | −27.8 ± 1.9 | 11.8 | 0.5 | |
PON2 | −25.9 | −26.1 ± 1.0 | 7.6 | 0 | |
His6-OPH | −23.0 | −22.6 ± 1.1 | 7.6 | 0.4 | |
Colistin | Acylase | −25.2 | −24.5 ± 1.4 | 7.2 | 0.2 |
Lactonase | −22.9 | −22.8 ± 1.4 | 12.9 | 0.5 | |
PON2 | −19.5 | −19.2 ± 0.8 | 16.3 | 0 | |
His6-OPH | −21.5 | −21.3 ± 1.0 | 9.6 | 0.1 | |
Oritavancin | Acylase | −41.6 | −41.8 ± 1.5 | 11.4 | 0.6 |
Lactonase | −35.5 | −34.9 ± 2.2 | 18.9 | 0.6 | |
PON2 | −29.4 | −29.1 ± 0.7 | 20.1 | 0 | |
His6-OPH | −33.7 | −33.5 ± 0.9 | 7.5 | 0.1 | |
Indolicidin | Acylase | −32.4 | −32.2 ± 1.0 | 10.2 | 0.5 |
Lactonase | −31.7 | −31.6 ± 1.2 | 15.1 | 0.7 | |
PON2 | −28.7 | −28.2 ± 1.0 | 19.5 | 0 | |
His6-OPH | −32.3 | −32.0 ± 1.2 | 7.8 | 0.6 | |
Temporin A | Acylase | −25.3 | −25.3 ± 0.4 | 9.2 | 0.6 |
Lactonase | −26.1 | −25.9 ± 0.6 | 15.8 | 0.7 | |
PON2 | −26.6 | −26.6 ± 0.5 | 16.5 | 0 | |
His6-OPH | −25.2 | −25.1 ± 0.2 | 9.6 | 0.1 |
G(+) Bacterial Cells:Bacillus subtilisB-522 | ||||
QQ Enzyme | Colistin | Polymyxin B | Indolicidin | Temporin A |
None | 6.16 ± 0.19 | 5.13 ± 0.16 | 66.72 ± 0.43 | 7.90 ± 0.06 |
His6-OPH | 1.41 ± 0.03 | 0.38 ± 0.01 | 12.24 ± 0.23 | 5.31 ± 0.01 |
Penicillin acylase | 0.47 ± 0.03 | 0.21 ± 0.01 | ||
Thermolysin | 614 ± 13 | 364 ± 12 | ||
Carboxypeptidase A | 14,166 ± 94 | 1684 ± 53 | ||
G(−) Bacterial Cells: Pseudomonas sp. 78G | ||||
None | 4.61 ± 0.22 | 4.35 ± 0.11 | 6.93 ± 0.18 | 2.24 ± 0.03 |
His6-OPH | 0.13 ± 0.02 | 1.16 ± 0.16 | 2.61 ± 0.24 | 1.18 ± 0.02 |
Penicillin acylase | 0.37 ± 0.02 | 0.97 ± 0.02 | ||
Thermolysin | 17.28 ± 0.49 | 3.14 ± 0.32 | ||
Carboxypeptidase A | 7.05 ± 0.19 | 33.28 ± 1.39 |
Peptidase (Source) [39] | Molar Mass; Optimal pH | * Preferable Peptide Bond for Cleavage |
---|---|---|
Coccolysin (Enterococcus faecalis) | 31.5 kDa; pH 6–8 | P1’ = Leu, Phe, Ile or Ala |
Griselysin (Streptomyces griseus) | 35.1 kDa; pH ~7 | P1, P1’ = hydrophobic residues |
Stearolysin (Geobacillus stearothermophilus) | 34 kDa; neutral pH | P1, P1’ = hydrophobic residues |
Mycolysin (Streptomyces sp.) | 37.1 kDa; pH 7–8.5 | P1’ = hydrophobic residues |
Microcystinase (Sphingomonadaceae sp.) | 36 kDa; pH 6.5–8.4 | Arg–Adda bond ** |
Carboxypeptidase A (bovine pancreas) | 34.6 kDa; pH 7–9 | C-terminal hydrophobic residues (incl. acylated ones) |
Antimicrobial Agent | Enzyme | Affinity, kJ × mol−1 | Occupied Area, % | ||
---|---|---|---|---|---|
Mean | Median | Total | Near Active Sites | ||
Dermicidin | Coccolysin | −14.4 | −14.2 ± 0.6 | 32.5 | 0.1 |
Griselysin | −13.1 | −13.0 ± 0.6 | 16.4 | 0 | |
Stearolysin | −16.4 | −16.3 ± 0.3 | 10.8 | 0 | |
Mycolysin | −12.3 | −12.3 ± 0.5 | 14.8 | 0.7 | |
Microcystinase | −13.4 | −13.4 ± 1.0 | 21.7 | 0 | |
Carboxypeptidase A | −18.7 | −18.4 ± 1.2 | 35.6 | 1.9 | |
Polymyxin B | Coccolysin | −29.1 | −29.1 ± 1.5 | 9.1 | 0.3 |
Griselysin | −30.5 | −30.3 ± 1.0 | 17.3 | 0.2 | |
Stearolysin | −35.0 | −35.1 ± 0.6 | 6.9 | 0 | |
Mycolysin | −32.3 | −32.2 ± 1.4 | 10.8 | 0.4 | |
Microcystinase | −26.8 | −26.8 ± 0.5 | 11.0 | 0.5 | |
Carboxypeptidase A | −26.3 | −26.6 ± 2.1 | 13.5 | 1.9 | |
Colistin | Coccolysin | −27.5 | −27.2 ± 1.4 | 9.5 | 0.3 |
Griselysin | −28.2 | −27.6 ± 1.9 | 18.8 | 0.2 | |
Stearolysin | −31.7 | −31.6 ± 1.2 | 5.4 | 0 | |
Mycolysin | −26.3 | −26.1 ± 0.6 | 12.6 | 0.5 | |
Microcystinase | −27.6 | −27.0 ± 2.1 | 10.3 | 0.6 | |
Carboxypeptidase A | −21.7 | −21.7 ± 0.9 | 7.6 | 1.9 | |
Oritavancin | Coccolysin | −39.0 | −38.5 ± 1.7 | 15.2 | 0.4 |
Griselysin | −39.0 | −38.9 ± 1.9 | 18.9 | 0.2 | |
Stearolysin | −47.4 | −47.1 ± 2.5 | 11.2 | 0.1 | |
Mycolysin | −34.7 | −34.3 ± 1.0 | 10.8 | 0.5 | |
Microcystinase | −38.0 | −37.6 ± 0.5 | 13.9 | 0.2 | |
Carboxypeptidase A | −36.7 | −36.6 ± 1.6 | 11.2 | 1.7 | |
Indolicidin | Coccolysin | −36.3 | −36.6 ± 1.1 | 12.0 | 0.4 |
Griselysin | −29.3 | −29.3 ± 0.9 | 18.6 | 0.3 | |
Stearolysin | −43.9 | −43.7 ± 0.8 | 10.2 | 0 | |
Mycolysin | −33.4 | −33.0 ± 0.9 | 6.9 | 0 | |
Microcystinase | −36.8 | −36.8 ± 0.8 | 13.0 | 0.6 | |
Carboxypeptidase A | −30.5 | −30.1 ± 1.3 | 14.0 | 1.9 | |
Temporin A | Coccolysin | −26.3 | −26.3 ± 1.4 | 9.8 | 0.3 |
Griselysin | −26.6 | −26.8 ± 0.4 | 11.4 | 0 | |
Stearolysin | −29.8 | −29.9 ± 0.6 | 9.2 | 0 | |
Mycolysin | −29.5 | −29.5 ± 1.6 | 8.7 | 0 | |
Microcystinase | −31.5 | −31.4 ± 1.1 | 11.6 | 0.6 | |
Carboxypeptidase A | −27.6 | −27.6 ± 0.8 | 12.1 | 1.9 |
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Aslanli, A.; Lyagin, I.; Stepanov, N.; Presnov, D.; Efremenko, E. Bacterial Cellulose Containing Combinations of Antimicrobial Peptides with Various QQ Enzymes as a Prototype of an “Enhanced Antibacterial” Dressing: In Silico and In Vitro Data. Pharmaceutics 2020, 12, 1155. https://doi.org/10.3390/pharmaceutics12121155
Aslanli A, Lyagin I, Stepanov N, Presnov D, Efremenko E. Bacterial Cellulose Containing Combinations of Antimicrobial Peptides with Various QQ Enzymes as a Prototype of an “Enhanced Antibacterial” Dressing: In Silico and In Vitro Data. Pharmaceutics. 2020; 12(12):1155. https://doi.org/10.3390/pharmaceutics12121155
Chicago/Turabian StyleAslanli, Aysel, Ilya Lyagin, Nikolay Stepanov, Denis Presnov, and Elena Efremenko. 2020. "Bacterial Cellulose Containing Combinations of Antimicrobial Peptides with Various QQ Enzymes as a Prototype of an “Enhanced Antibacterial” Dressing: In Silico and In Vitro Data" Pharmaceutics 12, no. 12: 1155. https://doi.org/10.3390/pharmaceutics12121155