Bacteriophage phi11 lysin: Physicochemical characterization and comparison with phage phi80α lysin

Filatova, L. Yu.; Donovan, David M.; Foster‐Frey, Juli; Pugachev, V. G.; Дмитриева, Н Ф; Chubar, T. A.; Klyachko, Natalia L.; Kabanov, Alexander V.

doi:10.1016/j.enzmictec.2015.03.005

Cited by 17 publications

(7 citation statements)

References 20 publications

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“…In this study, we demonstrated that PGHs selected for activity under relevant conditions and fused to CPPs are promising antibacterial agents that effectively eliminate intracellular S. aureus. The different, highly complex environments found inside and outside cellular compartments pose diverse challenges to PGH activity, as they vary in multiple parameters such as pH, ionic strength, and osmolality (36)(37)(38). To accommodate these challenges, we screened PGHs for high activity in multiple buffer systems that simulate conditions found in different intracellular niches of S. aureus, such as the cytoplasm or phagolysosomes.…”

Section: Discussionmentioning

confidence: 99%

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Röhrig

Huemer

Lorgé

et al. 2020

mBio

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Staphylococcus aureus is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of S. aureus plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan hydrolases (PGHs) are highly specific bactericidal enzymes active against both drug-sensitive and -resistant bacteria. However, PGHs able to effectively target intracellular S. aureus are not yet available. To overcome this limitation, we first screened 322 recombineered PGHs for staphylolytic activity under conditions found inside eukaryotic intracellular compartments. The most active constructs were modified by fusion to different cell-penetrating peptides (CPPs), resulting in increased uptake and enhanced intracellular killing (reduction by up to 4.5 log units) of various S. aureus strains (including methicillin-resistant S. aureus [MRSA]) in different tissue culture infection models. The combined application of synergistic PGH-CPP constructs further enhanced their intracellular efficacy. Finally, synergistically active PGH-CPP cocktails reduced the total S. aureus by more than 2.2 log units in a murine abscess model after peripheral injection. Significantly more intracellular bacteria were killed by the PGH-CPPs than by the PGHs alone. Collectively, our findings show that CPP-fused PGHs are effective novel protein therapeutics against both intracellular and drug-resistant S. aureus. IMPORTANCE The increasing prevalence of antibiotic-resistant bacteria is one of the most urgent problems of our time. Staphylococcus aureus is an important human pathogen that has acquired several mechanisms to evade antibiotic treatment. In addition, S. aureus is able to invade and persist within human cells, hiding from the immune response and antibiotic therapies. For these reasons, novel antibacterial strategies against these pathogens are needed. Here, we developed lytic enzymes which are able to effectively target drug-resistant and intracellular S. aureus. Fusion of these so-called enzybiotics to cell-penetrating peptides enhanced their uptake and intracellular bactericidal activity in cell culture and in an abscess mouse model. Our results suggest that cell-penetrating enzybiotics are a promising new class of therapeutics against staphylococcal infections.

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Section: Discussionmentioning

confidence: 99%

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Röhrig

Huemer

Lorgé

et al. 2020

mBio

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“…At temperatures over 40°C, a dramatic loss in stability was observed and attributed to secondarystructure changes, as measured by circular dichroism spectroscopy (134). The same group reported that LysK displayed increased stability in complexes with polycationic polymers (poly-L-lysines [PLLs]) and block copolymers of these PLLs with polyethylene glycol (135).…”

Section: Lysk and Its Homologues Structure Function And Physicochemmentioning

confidence: 99%

Recombinant Endolysins as Potential Therapeutics against Antibiotic-Resistant Staphylococcus aureus: Current Status of Research and Novel Delivery Strategies

et al. 2018

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is one of the most common pathogens of humans and animals, where it frequently colonizes skin and mucosal membranes. It is of major clinical importance as a nosocomial pathogen and causative agent of a wide array of diseases. Multidrug-resistant strains have become increasingly prevalent and represent a leading cause of morbidity and mortality. For this reason, novel strategies to combat multidrug-resistant pathogens are urgently needed. Bacteriophage-derived enzymes, so-called endolysins, and other peptidoglycan hydrolases with the ability to disrupt cell walls represent possible alternatives to conventional antibiotics. These lytic enzymes confer a high degree of host specificity and could potentially replace or be utilized in combination with antibiotics, with the aim to specifically treat infections caused by Gram-positive drug-resistant bacterial pathogens such as methicillin-resistant . LysK is one of the best-characterized endolysins with activity against multiple staphylococcal species. Various approaches to further enhance the antibacterial efficacy and applicability of endolysins have been demonstrated. These approaches include the construction of recombinant endolysin derivatives and the development of novel delivery strategies for various applications, such as the production of endolysins in lactic acid bacteria and their conjugation to nanoparticles. These novel strategies are a major focus of this review.

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“…Screening of phages-host interaction to increase lysis rates precisely (the kinetics exponential decay rate) with a functional lytic enzyme during phage infecting bacterial cells causes bacteriolysis and results in dead but intact bacterial cells (Filatova et al, 2015;Sun et al, 2015). Virolysins lysed cell wall peptidoglycan that yielded to primary amino sugars released and free intracellular components from hydrolyzed bacteria in the reaction qualitatively and quantitatively (Kadurugamuwa et al, 1998;Li et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…However, virolysins are promising enzybiotics for bacterial cell wall hydrolases Elshayeb et al 2015 even if these bacteria are resistant to lysozymes. They have narrow spectra of sensitive bacteria, minimizing the disturbance to normal microflora and a tremendous diversity of lytic bacteriophages in the biosphere, which guarantees their ability of targeting almost any bacteria (Filatova et al, 2015). To determine the kinetics exponential decay of virolysins mathematically, the resulting data points of equations were fit to curves solved on double reciprocal plot (Courchesne et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Potentiality of lytic bacteriophages and their virolysins in lysing multi-drug resistant Salmonella typhi

Elshayeb¹,

Ahmed²,

Siddig³

et al. 2017

Afr. J. Biotechnol.

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Bacteriophage virolysins or lytic enzymes are bacterial peptidoglycan hydrolases responsible for lysing bacterial cells. Consequently, they are used as enzybiotics alongside with bacteriophage therapy to remedy multi-drug resistant Salmonella typhi. The objective of this study was to evaluate the potentiality of lytic bacteriophages and their virolysins in curing multi-drug resistant S. typhi. S. typhi was isolated and identified according to WHO and ISO guidelines. Antibiotics susceptibilities were tested using CLSI recommendations. Correspondingly, bacteriophage-lysing efficiency was assayed by plaques formation using the double-layer agar technique. Virolysins were extracted using ultracentrifugation and purified by dialysis after buffering in ammonium sulfate. Virolysins activity was determined by measuring the reaction mixtures spectrophotometrically (bacteria incubated as substrate in 37°C for 4 h). The phages and virolysins kinetics exponential rates were calculated using specific differential equations. Susceptibility data plotted based on antibiogram criteria confirmed that 33% of S. typhi isolates were multi-drug resistant. For bacteriophage replication and multiplicity of infection, phages were amplified to produce the maximum particles of titers. The phage titration data fit on sonogram revealed exponential decay of S. typhi incubated for 12 h. Meanwhile, the enzyme kinetics exponential decay on double reciprocal plot showed irretrievable relationship of host decay in 4 h. Since phages depend on their lytic cycle in lysing bacterial host, their enzymes have more capability in decaying the host; besides they are safe and time-saving when used in the treatment of antibiotics resistant S. typhi.

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Bacteriophage phi11 lysin: Physicochemical characterization and comparison with phage phi80α lysin

Cited by 17 publications

References 20 publications

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant Staphylococcus aureus

Recombinant Endolysins as Potential Therapeutics against Antibiotic-Resistant Staphylococcus aureus: Current Status of Research and Novel Delivery Strategies

Potentiality of lytic bacteriophages and their virolysins in lysing multi-drug resistant Salmonella typhi

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