| Title: | Effect of peptides' binding on the antimicrobial activity and biocompatibility of protein-based substrates |
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| Authors: | ID Kaisersberger Vincek, Maja (Author)
ID Kokol, Vanja (Mentor) More about this mentor...  |
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| Files: |  DOK_Kaisersberger_Vincek_Maja_2017.pdf (3,98 MB)
MD5: 33BA98E9E6CA1CA5567FDCFD1B1A8ABE
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| Language: | English |
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| Work type: | Dissertation |
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| Typology: | 2.08 - Doctoral Dissertation |
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| Organization: | FS - Faculty of Mechanical Engineering
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| Abstract: | This work reveals the effect of coupling approach (chemical by using carbodiimide chemistry and grafting-to vs. grafting-from synthesis routes, and enzymatic by using transglutaminase) of a hydrophilic ε-poly-L-lysine (εPL) and an amphiphilic oligo-acyl-lysyl (OAK) derivative (K-7α12-OH) to wool fibers and gelatine (GEL) macromolecules, respectively, and substrates antibacterial activity against Gram-negative E. coli and Gram-positive S. aureus bacteria after 1–24 h of exposure, as well as their cytotoxicity. Different spectroscopic (ultraviolet-visible, infrared, fluorescence and electron paramagnetic resonance) and separation techniques (size-exclusion chromatography and capillary zone electrophoresis) as well as zeta potential and potentiometric titration analysis, were performed to confirm the covalent coupling of εPL/OAK, and to determine the amount and orientation of its immobilisation.
The highest and kinetically the fastest level of bacterial reduction was achieved with wool/GEL functionalised with εPL/OAK by chemical grafting-to approach. This effect correlated with both the highest grafting yield and conformationally the highly-flexible (brush-like) orientation linkage of εPL/OAK, implicating on the highest amount of accessible amino groups interacting with bacterial membrane. However, OAK`s amphipathic structure, the cationic charge and the hydrophobic moieties, resulted to relatively high reduction of S. aureus for grafting-from and the enzymatic coupling approaches using OAK-functionalised GEL.
The εPL/OAK-functionalised GEL did not induce toxicity in human osteoblast cells, even at ~25-fold higher concentration than bacterial minimum inhibitory (MIC) concentration of εPL/OAK, supporting their potential usage in biomedical applications.
It was also shown that non-ionic surfactant adsorbs strongly onto the wool surface during the process of washing, thereby blocking the functional sites of immobilized εPL and decreases its antibacterial efficiency.
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| Keywords: | wool, gelatine, antimicrobial peptides, ε-poly-L-lysine, oligo-acyl-lysyl, grafting chemistry, grafting approach, peptide orientation, antibacterial activity, cytotoxicity |
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| Place of publishing: | Maribor |
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| Publisher: | [M. Kaisersberger Vincek] |
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| Year of publishing: | 2017 |
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| PID: | 20.500.12556/DKUM-65459 |
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| UDC: | 677.31.017.86:604.4(043.3) |
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| COBISS.SI-ID: | 20730902 |
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| NUK URN: | URN:SI:UM:DK:H9R0AYPC |
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| Publication date in DKUM: | 17.08.2017 |
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| Views: | 2142 |
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| Downloads: | 237 |
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| Metadata: |    |
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| Categories: | KTFMB - FS
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