Effect of peptides' binding on the antimicrobial activity and biocompatibility of protein-based substrates Maja Kaisersberger Vincek
, 2017, doktorska disertacija
Opis: 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.
Ključne besede: wool, gelatine, antimicrobial peptides, ε-poly-L-lysine, oligo-acyl-lysyl, grafting chemistry, grafting approach, peptide orientation, antibacterial activity, cytotoxicity
Objavljeno: 17.08.2017; Ogledov: 111; Prenosov: 8
Polno besedilo (3,98 MB)
Antibacterial and anticandidal activity of Tylosema esculentum (marama) extractsWalter Chingwaru
, Gyebi Duodu
, Yolandi Van Zyl
, Runner Majinda
, Sam Yeboah
, Jose Jackson
, Petrina Kapewangolo
, Avrelija Cencič
, 2011, izvirni znanstveni članek
Opis: Bean and tuber extracts of Tylosema esculentum (marama) – an African creeping plant – were obtained using ethanol, methanol and water. Based on information that T. esculentum is used traditionally for the treatment of various diseases, the antibacterial and anticandidal effects of tuber and bean extracts were investigated. The antimicrobial activity of the extracts was tested on methicillin-resistant Staphylococcus aureus (MRSA, ATCC 6538), Mycobacterium terrae (ATCC 15755), Corynebacterium diphtheriae (clinical) and Candida albicans (ATCC 2091). We performed the broth microdilution test for the determination of the minimum inhibitory concentration (MIC) and a method to determine survival of microorganisms after in vitro co-incubation with the highest concentrations of T. esculentum extracts, followed by assessment of colony counts. Ethanol and methanol (phenolic) bean extracts exhibited higher potency against bacteria and yeast than aqueous extracts. Marama bean seed coat crude ethanolic extract (MSCE) and seed coat polyphenolic fractions, especially soluble-bound fraction (MSCIB), were highly antimicrobial against M. terrae, C. diphtheriae and C. albicans. All marama bean polyphenolic fractions, namely cotyledon acidified methanol fraction (MCAM), seed coat acidified methanol fraction (MSCAM), cotyledon insoluble-bound fraction (MCIB), seed coat insoluble-bound fraction (MSCIB), cotyledon-free polyphenolic fraction (MCFP) and seed coat free polyphenolic fraction (MSCFP) had high antimicrobial effects as shown by low respective MIC values between 0.1 mg/mL and 1 mg/mL. These MIC values were comparable to those of control antimicrobials used: amphotericin B (0.5 mg/mL) and cesfulodin (0.1 mg/mL) against C. diphtheriae, streptomycin (1.0 mg/mL) and gentamicin (0.4 mg/mL) against M. terrae, and amphotericin B (0.05 mg/mL) against C. albicans. Marama seed coat soluble-esterified fraction (MSCS) had closer activity to that of cefsulodin against M. terrae. High amounts of phenolic substances, such as gallic acid, especially in the seed coats, as well as high amounts of phytosterols, lignans, certain fatty acids and peptides (specifically protease inhibitors) in the cotyledons contributed to the observed antibacterial and anticandidal activities. Marama extracts, especially phenolic and crude seed coat extracts, had high multi-species antibacterial and anticandidal activities at concentrations comparable to that of some conventional drugs; these extracts have potential use as microbicides.
Ključne besede: marama, Tylosema esculentum, antibacterial activity, anticandidal activity
Objavljeno: 07.08.2017; Ogledov: 52; Prenosov: 3
Polno besedilo (1,38 MB)