scholarly journals Mode of Action of the Antimicrobial Peptide Aureocin A53 from Staphylococcus aureus

2002 ◽  
Vol 68 (11) ◽  
pp. 5274-5280 ◽  
Author(s):  
Daili Jacqueline Aguilar Netz ◽  
Maria do Carmo de Freire Bastos ◽  
Hans-Georg Sahl
Keyword(s):  
Mode Of Action ◽  
Neutral Lipids ◽  
Membrane Damage ◽  
Model Membranes ◽  
Bacterial Cells ◽  
Gram Staining ◽  
Rapid Release ◽  
High Concentrations ◽  
Staphylococcus Simulans ◽  
Tryptophan Emission

ABSTRACT We investigated the mode of action of aureocin A53 on living bacterial cells and model membranes. Aureocin A53 acted bactericidally against Staphylococcus simulans 22, with >90% of the cells killed within a few minutes. Cell death was followed by lysis, as indicated by a clearing of the cell suspension and Gram staining. Aureocin A53 rapidly dissipated the membrane potential and simultaneously stopped biosynthesis of DNA, polysaccharides, and protein. Aureocin A53 induced a rapid release of preaccumulated glutamate and Rb+. Experiments on model membranes demonstrated that aureocin A53 provoked significant leakage of carboxyfluorescein (CF) exclusively from acidic liposomes but only at relatively high concentrations (0.5 to 8 mol%). Thus, the bactericidal activity of aureocin A53 derives from membrane permeation via generalized membrane destruction rather than by formation of discrete pores within membranes. Tryptophan emission fluorescence spectroscopy demonstrated interaction of aureocin A53 with both acidic and neutral membranes, as indicated by similar blue shifts. Since there was no significant aureocin A53-induced CF leakage from neutral liposomes, its appears that the peptide does interact with neutral lipids without provoking membrane damage.

scholarly journals Mode-of-Action of Antimicrobial Peptides: Membrane Disruption vs. Intracellular Mechanisms

2020 ◽  
Vol 2 ◽  
Author(s):  
Aurélie H. Benfield ◽  
Sónia Troeira Henriques
Keyword(s):  
Antimicrobial Peptides ◽  
Physicochemical Properties ◽  
Broad Spectrum ◽  
Mode Of Action ◽  
Bacterial Pathogens ◽  
Model Membranes ◽  
Membrane Disruption ◽  
Attractive Alternative ◽  
Bacterial Cells

Antimicrobial peptides are an attractive alternative to traditional antibiotics, due to their physicochemical properties, activity toward a broad spectrum of bacteria, and mode-of-actions distinct from those used by current antibiotics. In general, antimicrobial peptides kill bacteria by either disrupting their membrane, or by entering inside bacterial cells to interact with intracellular components. Characterization of their mode-of-action is essential to improve their activity, avoid resistance in bacterial pathogens, and accelerate their use as therapeutics. Here we review experimental biophysical tools that can be employed with model membranes and bacterial cells to characterize the mode-of-action of antimicrobial peptides.

scholarly journals The Mode of Action of Cyclic Monoterpenes (−)-Limoneneand (+)-α-Pinene on Bacterial Cells

Biomolecules ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 806
Author(s):  
Olga E. Melkina ◽  
Vladimir A. Plyuta ◽  
Inessa A. Khmel ◽  
Gennadii B. Zavilgelsky
Keyword(s):  
Heat Shock ◽  
Biological Activities ◽  
Membrane Damage ◽  
Genetically Engineered ◽  
Bacterial Cells ◽  
Prolonged Incubation ◽  
E Coli ◽  
Volatile Organic ◽  
High Concentrations ◽  
First Time

A broad spectrum of volatile organic compounds’ (VOCs’) biological activities has attracted significant scientific interest, but their mechanisms of action remain little understood. The mechanism of action of two VOCs—the cyclic monoterpenes (−)-limonene and (+)-α-pinene—on bacteria was studied in this work. We used genetically engineered Escherichia coli bioluminescent strains harboring stress-responsive promoters (responsive to oxidative stress, DNA damage, SOS response, protein damage, heatshock, membrane damage) fused to the luxCDABE genes of Photorhabdus luminescens. We showed that (−)-limonene induces the PkatG and PsoxS promoters due to the formation of reactive oxygen species and, as a result, causes damage to DNA (SOSresponse), proteins (heat shock), and membrane (increases its permeability). The experimental data indicate that the action of (−)-limonene at high concentrations and prolonged incubation time makes degrading processes in cells irreversible. The effect of (+)-α-pinene is much weaker: it induces only heat shock in the bacteria. Moreover, we showed for the first time that (−)-limonene completely inhibits the DnaKJE–ClpB bichaperone-dependent refolding of heat-inactivated bacterial luciferase in both E. coli wild type and mutant ΔibpB strains. (+)-α-Pinene partially inhibits refolding only in ΔibpB mutant strain.

scholarly journals Effect of Pseudomonas moorei KB4 Cells’ Immobilisation on Their Degradation Potential and Tolerance towards Paracetamol

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 820
Author(s):  
Robert Surma ◽  
Danuta Wojcieszyńska ◽  
Jagna Karcz ◽  
Urszula Guzik
Keyword(s):  
Protective Effect ◽  
Kinetic Parameters ◽  
Toxicity Assessment ◽  
Bacterial Cells ◽  
Low Concentrations ◽  
Toxicity Analysis ◽  
Immobilised Cells ◽  
Degradation Activity ◽  
High Concentrations ◽  
The Hill

Pseudomonas moorei KB4 is capable of degrading paracetamol, but high concentrations of this drug may cause an accumulation of toxic metabolites. It is known that immobilisation can have a protective effect on bacterial cells; therefore, the toxicity and degradation rate of paracetamol by the immobilised strain KB4 were assessed. Strain KB4 was immobilised on a plant sponge. A toxicity assessment was performed by measuring the concentration of ATP using the colony-forming unit (CFU) method. The kinetic parameters of paracetamol degradation were estimated using the Hill equation. Toxicity analysis showed a protective effect of the carrier at low concentrations of paracetamol. Moreover, a pronounced phenomenon of hormesis was observed in the immobilised systems. The obtained kinetic parameters and the course of the kinetic curves clearly indicate a decrease in the degradation activity of cells after their immobilisation. There was a delay in degradation in the systems with free cells without glucose and immobilised cells with glucose. However, it was demonstrated that the immobilised systems can degrade at least ten succeeding cycles of 20 mg/L paracetamol degradation. The obtained results indicate that the immobilised strain may become a useful tool in the process of paracetamol degradation.

scholarly journals Adhesive properties of an outer structure of Clostridium perfringens type A isolated from piglets with with catarrhal enteritis

1999 ◽  
Vol 30 (3) ◽  
pp. 242-248 ◽  
Author(s):  
Elizabeth Pelosi Teixeira ◽  
Marlene Braide Serafim ◽  
Maria Alice Cruz Höfling ◽  
Aureo T. Yamada ◽  
Antonio Fernando Pestana de Castro
Keyword(s):  
Clostridium Perfringens ◽  
Type A ◽  
Negative Control ◽  
Bacterial Cells ◽  
Intestinal Epithelial ◽  
Adhesive Properties ◽  
Ileal Loop ◽  
Fibrillar Material ◽  
Gram Staining ◽  
Transmission Electron

One strain (S32) of Clostridium perfringens type A was isolated from a case of catarrhal enteritis of piglets. This strain was able to adhere to HeLa cells showing an adherence index (AI) of 25.15 ± 1.26 (mean ± 1 standard error of the mean). Treatment of the bacterial cells with trypsin (0.25mg/ml) decreased in 70%-80% the AI and metaperiodate (10mg/ml) abolished completely the adherence, suggesting that the structure responsible for this phenomenon was probably a glycoprotein. Heating of bacterial suspensions (100ºC/5 min) before carrying out the adhesion test decreased the AI rendering it equal to the negative controls. Rabbit homologous S32 antiserum inhibited the adherence up to dilutions of 1: 640, at least. The piglet ileal loop assay, carried out with strains S32 and Jab-1 (negative control) demonstrated that the strain S32 was able to adhere to the intestinal epithelial cells when examined after Gram staining. Transmission electron microcopy (TEM) demonstrated that S32 strain displayed a loose fibrillar material not seen with Jab-1. Stabilization of the bacterial cells with homologous antiserum of strain S32, followed by staining with rhuteniun red, revealed loose long fibrillar material on the outer surface of the cells, that sometimes could be seen spreading out from the cells and linking bacterial cells. The question whether this structure might be an adhesin for this strain of Cl. perfringes type A, perhaps playing a role in the pathogenesis of the catarrhal enteritis of piglets, is dependent on further studies.

scholarly journals Plant defensin antibacterial mode of action against Pseudomonas species

2020 ◽  
Author(s):  
Andrew Edward Sathoff ◽  
Shawn Lewenza ◽  
Deborah A. Samac
Keyword(s):  
Gene Expression ◽  
Antibacterial Activity ◽  
Outer Membrane ◽  
Pseudomonas Syringae ◽  
Mode Of Action ◽  
Fluorescent Microscopy ◽  
Insertion Site ◽  
Membrane Damage ◽  
Plant Defensin ◽  
Plant Defensins

Abstract Background: Though many plant defensins exhibit antibacterial activity, little is known about their antibacterial mode of action (MOA). Antimicrobial peptides with a characterized MOA induce the expression of multiple bacterial outer membrane modifications, which are required for resistance to these membrane-targeting peptides. Mini-Tn5-lux mutant strains of Pseudomonas aeruginosa with Tn insertions disrupting outer membrane protective modifications were assessed for sensitivity against plant defensin peptides. These transcriptional lux reporter strains were also evaluated for lux gene expression in response to sublethal plant defensin exposure. Also, a plant pathogen, Pseudomonas syringae pv. syringae was modified through transposon mutagenesis to create mutants that are resistant to in vitro MtDef4 treatments.Results: Plant defensins displayed specific and potent antibacterial activity against strains of P. aeruginosa. A defensin from Medicago truncatula, MtDef4, induced dose-dependent gene expression of the aminoarabinose modification of LPS and surface polycation spermidine production operons. The ability for MtDef4 to damage bacterial outer membranes was also verified visually through fluorescent microscopy. Another defensin from M. truncatula, MtDef5, failed to induce lux gene expression and limited outer membrane damage was detected with fluorescent microscopy. The transposon insertion site on MtDef4 resistant P. syringae pv. syringae mutants was sequenced, and modifications of ribosomal genes were identified to contribute to enhanced resistance to plant defensin treatments. Conclusions: MtDef4 damages the outer membrane similar to polymyxin B, which stimulates antimicrobial peptide resistance mechanisms to plant defensins. MtDef5, appears to have a different antibacterial MOA. Additionally, the MtDef4 antibacterial mode of action may also involve inhibition of translation.

scholarly journals The amphibian antimicrobial peptide uperin 3.5 is a cross-α/cross-β chameleon functional amyloid

2021 ◽  
Vol 118 (3) ◽  
pp. e2014442118
Author(s):  
Nir Salinas ◽  
Einav Tayeb-Fligelman ◽  
Massimo D. Sammito ◽  
Daniel Bloch ◽  
Raz Jelinek ◽  
...  
Keyword(s):  
Antimicrobial Peptide ◽  
Amyloid Fibrils ◽  
Amyloid Fibril ◽  
Membrane Damage ◽  
Fibril Formation ◽  
Regulation Mechanism ◽  
Bacterial Cells ◽  
Amyloid Fibril Formation ◽  
Β Amyloid ◽  
Β Sheets

Antimicrobial activity is being increasingly linked to amyloid fibril formation, suggesting physiological roles for some human amyloids, which have historically been viewed as strictly pathological agents. This work reports on formation of functional cross-α amyloid fibrils of the amphibian antimicrobial peptide uperin 3.5 at atomic resolution, an architecture initially discovered in the bacterial PSMα3 cytotoxin. The fibrils of uperin 3.5 and PSMα3 comprised antiparallel and parallel helical sheets, respectively, recapitulating properties of β-sheets. Uperin 3.5 demonstrated chameleon properties of a secondary structure switch, forming mostly cross-β fibrils in the absence of lipids. Uperin 3.5 helical fibril formation was largely induced by, and formed on, bacterial cells or membrane mimetics, and led to membrane damage and cell death. These findings suggest a regulation mechanism, which includes storage of inactive peptides as well as environmentally induced activation of uperin 3.5, via chameleon cross-α/β amyloid fibrils.

Revealing the Mode of Action of Halictine Antimicrobial Peptides: A Comprehensive Study with Model Membranes

Langmuir ◽  
2020 ◽  
Vol 36 (19) ◽  
pp. 5145-5155 ◽  
Author(s):  
Tatiana M. Domingues ◽  
Katia R. Perez ◽  
Karin A. Riske
Keyword(s):  
Antimicrobial Peptides ◽  
Mode Of Action ◽  
Model Membranes ◽  
Comprehensive Study

scholarly journals Submicellar complexes may initiate the fungicidal effects of cationic amphiphilic compounds on Candida albicans.

1997 ◽  
Vol 41 (3) ◽  
pp. 544-550 ◽  
Author(s):  
B Ahlström ◽  
M Chelminska-Bertilsson ◽  
R A Thompson ◽  
L Edebo
Keyword(s):  
Candida Albicans ◽  
Membrane Damage ◽  
Hydrocarbon Chain ◽  
Hexadecyltrimethylammonium Bromide ◽  
Tetradecyltrimethylammonium Bromide ◽  
Low Concentrations ◽  
Hydrocarbon Chains ◽  
Absorbing Material ◽  
High Concentrations ◽  
Chain Lengths

The killing of Candida albicans by a series of amphiphilic quaternary ammonium compounds (QACs) with different hydrocarbon chain lengths was closely related to the binding of the compounds to the cells and damage of the cell membranes. The membrane damage was measured as the level of release of the UV-absorbing material into the medium in which the cells were suspended and as the level of uptake of propidium iodide in individual cells by flow cytometry. It was shown that of the compounds tested, hexadecyltrimethylammonium bromide (cetyltrimethylammonium bromide [CTAB]) bound most efficiently. Tetradecyl betainate chloride (B14), tetradecanoylcholine bromide (C14), tetradecyltrimethylammonium bromide (TTAB), and dodecyltrimethylammonium bromide (DTAB) followed and had declining degrees of binding efficiency. The proportion of CTAB bound was almost total at concentrations up to the critical micelle concentration (CMC) of the compound, whereas that of B14 was somewhat smaller. For the two remaining tetradecyl compounds (C14 and TTAB), still smaller proportions were bound at low concentrations, but the proportions rose disproportionally at increasing concentrations to a distinct maximum at concentrations of 0.2 to 0.5 times the CMC. We propose that interfacial micelle-like aggregates are formed at the cell surface as a step in the binding process. An analogous, but less conspicuous, maximum was seen for DTAB. Thus, great differences in the binding affinity of QACs with different hydrocarbon chains at different concentrations to C. albicans were observed. These differences were related to the CMC of the compound. In contrast, the binding of TTAB to Salmonella typhimurium 395 MS was almost total at low as well as high concentrations until saturation was attained, indicating fundamental differences between binding to the yeast and binding to gram-negative bacteria. The importance of lipid-type complexes or aggregates to the antifungal effect of membrane-active substances are discussed.

Control of Fatty Acid and Lipid Biosynthesis in Euglena gracilis by Ammonia, Light and DCMU

1972 ◽  
Vol 27 (1) ◽  
pp. 53-61 ◽  
Author(s):  
P. Pohl ◽  
H. Wagner
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
White Light ◽  
Euglena Gracilis ◽  
Fatty Acid Biosynthesis ◽  
Neutral Lipids ◽  
Fluorescent Light ◽  
Low Levels ◽  
High Concentrations ◽  
Monogalactosyl Diglyceride

When Euglena gracilis was grown under white (fluorescent) light in media containing high concentrations of ammonium chloride (more than 0.005%), the main lipids synthesized were monogalactosyl diglyceride, digalactosyl diglyceride, phosphatidyl glycerol, sulfolipid and the all cis △7,10-16:2, △7,10,13-16:3, △4,7,10,13-16:4, △9,12-18:2 and △9,12,15-18:3 fatty acids. At low levels of ammonium (less than 0.002%) these compounds were produced only in small amounts, while neutral lipids, phosphatidyl choline, phosphatidyl ethanolamine and the 14:0, 16:0 and 16:1 fatty acids predominated.When Euglena gracilis was grown in white light in the presence of dichlorophenyldimethylurea (DCMU) or in the dark, fatty acid and lipid biosyntheses followed the same pattern as in white light at low levels of ammonium. Similar results were obtained when nitrate served as the only nitrogen source in the light and in the dark.The results indicate that in Euglena gracilis there are a light independent and a light and ammonium dependent pathway of fatty acid biosynthesis. Both pathways seem to be in association with specific lipids.

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