Atomic force microscopy and surface plasmon resonan<b>ce for real-time single-cell monitoring of bacteriophage-</b>mediated lysis of bacteria

Obořilová, Radka; Šimečková, Hana; Pastucha, Matěj; Klimovič, Šimon; Víšová, Ivana; Přibyl, Jan; Vaisocherová‐Lísalová, Hana; Pantůček, Roman; Skládal, Petr; Mašlaňová, Ivana

doi:10.1039/d1nr02921e

Cited by 6 publications

(2 citation statements)

References 63 publications

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“…The lysin-biofilm removal process is, however, indistinguishable but evident in the curve (i.e., a weak signal resulting in a 6-degree decline from 20 to 35 min). This decline may be caused by lysed bacterial clusters that are still covered on the SPR chip as previously reported ( Obořilová et al, 2021 ) and observed by SEM in Supplementary Figure 2 . In the SEM result, further lysis results in destruction of the biofilm which may characterize stage III of the sensogram.…”

Section: Discussionsupporting

confidence: 74%

In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance

Hong

Nyaruaba

et al. 2021

Front. Microbiol.

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Staphylococcus aureus can produce a multilayered biofilm embedded in extracellular polymeric matrix. This biofilm is difficult to remove, insensitive to antibiotics, easy to develop drug-resistant strains and causes enormous problems to environments and health. Phage lysin which commonly consists of a catalytic domain (CD) and a cell-wall binding domain (CBD) is a powerful weapon against bacterial biofilm. However, the real-time interaction between lysin and S. aureus biofilm is still not fully understood. In this study, we monitored the interactions of three lysins (ClyF, ClyC, PlySs2) against culture-on-chip S. aureus biofilm, in real-time, based on surface plasmon resonance (SPR). A typical SPR response curve showed that the lysins bound to the biofilm rapidly and the biofilm destruction started at a longer time. By using 1:1 binding model analysis, affinity constants (KD) for ClyF, ClyC, and PlySs2 were found to be 3.18 ± 0.127 μM, 1.12 ± 0.026 μM, and 15.5 ± 0.514 μM, respectively. The fact that ClyF and PlySs2 shared the same CBD but showed different affinity to S. aureus biofilm suggested that, not only CBD, but also CD affects the binding activity of the entire lysin. The SPR platform can be applied to improve our understanding on the complex interactions between lysins and bacterial biofilm including association (adsorption) and disassociation (destruction).

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Section: Discussionsupporting

confidence: 74%

In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance

Hong

Nyaruaba

et al. 2021

Front. Microbiol.

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“…The enzyme kills a wide range of clinical isolates of S. aureus , including MRSA, − and lyses both coagulase-positive and coagulase-negative strains . Using SEM and AFM analysis, it is clear that lysostaphin treatment causes nanoscale perforation, formation of spheroplasts and protoplasts, followed by cell swelling and complete cell lysis (Figure B). − This enzyme is active against cell wall fragments, heat-inactivated cells or viable cells regardless of their metabolic state − or encapsulation conditions. , In addition, lysostaphin can disrupt the biofilms of S. aureus , although the effect varies with conditions used for biofilm formation. ,− This enzyme is unable to kill intracellular S. aureus due to its inability to enter mammalian cells, ,− which is nonetheless challenged by a very recent study showing that lysostaphin alone killed intracellular S. aureus to a great extent in several cell lines in a dose-dependent manner …”

Section: Introductionmentioning

confidence: 99%

Lysostaphin: Engineering and Potentiation toward Better Applications

Zha

Zheng

et al. 2022

J. Agric. Food Chem.

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Lysostaphin is a potent bacteriolytic enzyme with endopeptidase activity against the common pathogen Staphylococcus aureus. By digesting the pentaglycine crossbridge in the cell wall peptidoglycan of S. aureus including the methicillin-resistant strains, lysostaphin initiates rapid lysis of planktonic and sessile cells (biofilms) and has great potential for use in agriculture, food industries, and pharmaceutical industries. In the past few decades, there have been tremendous efforts in potentiating lysostaphin for better applications in these fields, including engineering of the enzyme for higher potency and lower immunogenicity with longer-lasting effects, formulation and immobilization of the enzyme for higher stability and better durability, and recombinant expression for low-cost industrial production and in situ biocontrol. These achievements are extensively reviewed in this article focusing on applications in disease control, food preservation, surface decontamination, and pathogen detection. In addition, some basic properties of lysostaphin that have been controversial and only elucidated recently are summarized, including the substrate-binding properties, the number of zinc-binding sites, the substrate range, and the cleavage site in the pentaglycine crossbridge. Resistance to lysostaphin is also highlighted with a focus on various mechanisms. This article is concluded with a discussion on the limitations and future perspectives for the actual applications of lysostaphin.

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Morphological Characterization of Starches

Liu,

Jia,

Chen

et al. 2024

Methods and Protocols in Food Science

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Atomic force microscopy and surface plasmon resonance for real-time single-cell monitoring of bacteriophage-mediated lysis of bacteria

Abstract: The growing incidence of multidrug-resistant bacterial strains presents a major challenge in modern medicine. Antibiotic resistance is often exhibited by Staphylococcus aureus, which causes severe infections in human and animal...

Cited by 6 publications

References 63 publications

In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance

In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance

Lysostaphin: Engineering and Potentiation toward Better Applications

Morphological Characterization of Starches

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