Characterization of a novel endolysin LysSA11 and its utility as a potent biocontrol agent against Staphylococcus aureus on food and utensils

Chang, Yoonjee; Kim, Minsik; Ryu, Sangryeol

doi:10.1016/j.fm.2017.07.004

Cited by 73 publications

(48 citation statements)

References 40 publications

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“…The endolysin E-LM12 demonstrated capability to lyse all the Staphylococcus strains tested, including CoNS. These results are in agreement with other studies that report that Staphylococcus phage endolysins present a broad lytic activity covering all the Staphylococcus genus and commonly their spectrum is similar to the one from their CBD 56,63 . Also, the amino acid sequence of E-LM12 is very conserved, being closely related to several staphylococcal endolysins, namely LysK, which was described to be effective against S. aureus and CoNS 64 .…”

Section: Discussionsupporting

confidence: 93%

A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood

Costa

Dias

Melo

et al. 2020

Sci Rep

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Bloodstream infections (BSIs) are considered a major cause of death worldwide. Staphylococcus spp. are one of the most BSIs prevalent bacteria, classified as high priority due to the increasing multidrug resistant strains. Thus, a fast, specific and sensitive method for detection of these pathogens is of extreme importance. In this study, we have designed a novel assay for detection of Staphylococcus in blood culture samples, which combines the advantages of a phage endolysin cell wall binding domain (CBD) as a specific probe with the accuracy and high-throughput of flow cytometry techniques. In order to select the biorecognition molecule, three different truncations of the C-terminus of Staphylococcus phage endolysin E-LM12, namely the amidase (AMI), SH3 and amidase+SH3 (AMI_SH3) were cloned fused with a green fluorescent protein. From these, a higher binding efficiency to Staphylococcus cells was observed for AMI_SH3, indicating that the amidase domain possibly contributes to a more efficient binding of the SH3 domain. The novel phage endolysin-based flow cytometry assay provided highly reliable and specific detection of 1-5 CFU of Staphylococcus in 10 mL of spiked blood, after 16 hours of enrichment culture. Overall, the method developed herein presents advantages over the standard BSIs diagnostic methods, potentially contributing to an early and effective treatment of BSIs. Bloodstream infections (BSIs) are severe diseases caused by the presence of microorganisms, mainly bacteria, in blood and are characterized by high morbidity and mortality 1,2. The BSIs and associated organ dysfunctions (sepsis or septic shock) remain a life-threating disease due, partially, to the inability to rapidly detect and identify the responsible pathogens 3,4. Staphylococcus spp. are Gram-positive facultative anaerobic bacteria that frequently colonize the human body 5,6. These pathogens are becoming increasingly resistant to antibiotics and are well-established in both community and healthcare environments, being commonly isolated in intensive care units (ICU) 6,7. Staphylococcus aureus is a common cause of a variety of infections, from superficial skin infections to life-threatening diseases, including necrotizing pneumonia 8 , infective endocarditis 9 and BSIs 10. Coagulase-negative staphylococci (CoNS) have also been described as harmful to humans, causing several infections, particularly in patients with implanted medical devices 6. The empirical antibiotic therapy remains the standard of BSIs treatments 11 and its correct use within the first hour after the recognition of the BSI is recommended by the Surviving Sepsis Campaign Guidelines 11 and was reported as having a great impact on the patient survival rate 12. Nevertheless, the extensive use of broad-spectrum antibiotics and the large number of patients having negative blood culture samples and thus receiving unnecessary antibiotic treatment, are important contributors to the increase of antimicrobial resistance 13-15. Thus, sensitive, rapid, cost-efficient and specific ...

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Section: Discussionsupporting

confidence: 93%

A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood

Costa

Dias

Melo

et al. 2020

Sci Rep

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“…The lytic activity was calculated after 40 min as follows: {ΔOD 600 test (endolysin added)-ΔOD 600 control (buffer only)}/initial OD 600 . Antimicrobial spectrum was tested by plate lysis assay as previously described [ 16 ]. In brief, 10 μL of diluted endolysin in reaction buffer (32 μM; final concentration) was spotted onto a freshly prepared bacterial lawn on BHI agar plates.…”

Section: Methodsmentioning

confidence: 99%

Clostridium perfringens Virulent Bacteriophage CPS2 and Its Thermostable Endolysin LysCPS2

Son

Ryu

2018

Viruses

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Clostridium perfringens is one of the most common causes of food-borne illness. The increasing prevalence of multidrug-resistant bacteria requires the development of alternatives to typical antimicrobial treatments. Here, we isolated and characterized a C. perfringens-specific virulent bacteriophage CPS2 from chicken feces. The CPS2 phage contains a 17,961 bp double-stranded DNA genome with 25 putative ORFs, and belongs to the Picovirinae, subfamily of Podoviridae. Bioinformatic analysis of the CPS2 genome revealed a putative endolysin, LysCPS2, which is homologous to the endolysin of Clostridium phage phiZP2 and phiCP7R. The enzyme showed strong lytic activity against C. perfringens with optimum conditions at pH 7.5–10, 25–65 °C, and over a broad range of NaCl concentrations. Interestingly, LysCPS2 was found to be highly thermostable, with up to 30% of its lytic activity remaining after 10 min of incubation at 95 °C. The cell wall binding domain in the C-terminal region of LysCPS2 showed a binding spectrum specific to C. perfringens strains. This is the first report to characterize highly thermostable endolysin isolated from virulent C. perfringens bacteriophage. The enzyme can be used as an alternative biocontrol and detection agent against C. perfringens.

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“…Based on the above observations, depolymerase activity may have remained functional at 4 C in order to support bacteriophage infection. Previously, a well-characterized bacteriophage endolysin has demonstrated activity at refrigeration temperature (4 C; Chang, Kim, & Ryu, 2017), however, confirmation of depolymerase activity at this temperature would require further investigation. To the best of our knowledge, this is the first study to examine the efficacy of bacteriophage (with low MOI of 10 2 ) to control C. jejuni on refrigerated mutton and chicken meat in Malaysia.…”

Section: Bacteriophage Application In Mutton and Chicken Meatmentioning

confidence: 99%

Partial characterization and in vitro evaluation of a lytic bacteriophage for biocontrol of Campylobacter jejuni in mutton and chicken meat

Thung

Lee

Mahyudin

et al. 2020

Journal of Food Safety

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Campylobacter jejuni is a major foodborne pathogen causing acute bacterial enteritis worldwide. Screening of novel bacteriophages as biocontrol tools to eliminate C. jejuni in food during pre‐ and postharvest is an emerging research field. In this study, we partially characterized C. jejuni bacteriophage CJ01 for potential use as biocontrol agent. Transmission electron microscopy revealed bacteriophage CJ01 belonged to the Myoviridae family with a genome size of approximately 136 kb. The results of host range assay revealed CJ01 was able to infect C. jejuni and C. lari strains, with burst size about 35 phages per infected cell and latent period of 30 min. The effectiveness of bacteriophage CJ01 (with 100 multiplicity of infection = 100) was determined in mutton and chicken meat artificially contaminated with C. jejuni. The amount of C. jejuni in treated mutton and chicken meat were significantly reduced by 1.70 log CFU/g and 1.68 log CFU/g, respectively, after 48 hr stored at refrigeration temperature (4°C). The data presented here suggest that bacteriophage CJ01 can be used as a postharvest biocontrol agent.

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Characterization of a novel endolysin LysSA11 and its utility as a potent biocontrol agent against Staphylococcus aureus on food and utensils

Cited by 73 publications

References 40 publications

A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood

A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood

Clostridium perfringens Virulent Bacteriophage CPS2 and Its Thermostable Endolysin LysCPS2

Partial characterization and in vitro evaluation of a lytic bacteriophage for biocontrol of Campylobacter jejuni in mutton and chicken meat

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