Characterization of the Bacteriophage-Derived Endolysins PlySs2 and PlySs9 with In Vitro Lytic Activity against Bovine Mastitis Streptococcus uberis

Elst, Niels Vander; Linden, Sara B.; Lavigne, Rob; Meyer, Evelyne; Briers, Yves; Nelson, Daniel

doi:10.3390/antibiotics9090621

Cited by 27 publications

(52 citation statements)

References 42 publications

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“…Turbidity reduction assay was carried out according to Vander Elst et al 44 with some modifications. Briefly, Escherichia coli C, C600 (K-12), HB101 and B/R strains were grown in separate tubes overnight in Luria Broth at 37 °C in the air shaker.…”

Section: Methodsmentioning

confidence: 99%

Substantiation of propitious “Enzybiotic” from two novel bacteriophages isolated from a wastewater treatment plant in Qatar

Ramadoss

Al-Shukri

Shomar

et al. 2022

Sci Rep

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Lysin of bacteriophages isolated from a particular ecosystem could be inducted as a bio-controlling tool against the inhabiting pathogenic bacterial strains. Our study aims at both experimental and computational characterization of the identical lysin gene product inherent in the genomes of two novel Myoviridae bacteriophages, Escherichia Phage C600M2 (GenBank accession number OK040807, Protein ID: UCJ01465) and Escherichia Phage CL1 (GenBank Genome accession number OK040806.1, Protein ID: UCJ01321) isolated from wastewater collected from the main water treatment plant in Qatar. The lysin protein, evinced to be a globular N-acetyl-muramidase with intrinsic “cd00737: endolysin_autolysin” domain, was further expressed and purified to be experimentally validated by turbidimetric assay for its utility as an anti-bacterial agent. Comprehensive computational analysis revealed that the scrutinized lysin protein shared 85–98% sequence identity with 61 bacteriophages, all native to wastewater allied environments. Despite varied Host Recognition Components encoded in their genomes, the similitude of lysins, suggests its apparent significance in host–pathogen interactions endemic to wastewater environment. The present study substantiates the identical lysin from Escherichia Phage C600M2 and Escherichia Phage CL1 as propitious “enzybiotic”, a hybrid term to describe enzymes analogous to anti-biotics to combat antibiotic-resistant bacteria by in silico analysis and subsequent experimental validation.

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Section: Methodsmentioning

confidence: 99%

Substantiation of propitious “Enzybiotic” from two novel bacteriophages isolated from a wastewater treatment plant in Qatar

Ramadoss

Al-Shukri

Shomar

et al. 2022

Sci Rep

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“…Such candidate gene products were cloned into E. coli for expression and tested for their lytic activity in vitro and in vivo . This approach was used to obtain the PlyPy endolysin from S. pyogenes , PlySK1249 from S. dysgalactiae , λSa1 and λSa2 from S. agalactiae , Skl from S. mitis , and PlySs2 and PlySs9 from S. suis serotypes 2 and 9 ( Nelson et al, 2001 ; Llull et al, 2006 ; Gilmer et al, 2013 ; Oechslin et al, 2013 ; Lood et al, 2014 ; Vander Elst et al, 2020 ). An overall classification of streptococcal endolysins reported to date and their corresponding streptococcal species targets are summarized in Figure 2 .…”

Section: Sources and Isolation Of Streptococcal Endolysinsmentioning

confidence: 99%

“…However, some endolysins have an EAD at both N- and C-terminal domains with their CBD in the middle of the protein ( Lood et al, 2014 ). Several endolysins hold more than one EAD, such as amidase, endopeptidase, and glycosidase domains, while others only have one EAD ( Pritchard et al, 2007 ; McGowan et al, 2012 ; Vander Elst et al, 2020 ).…”

Section: Enzymatically Active Domain and Cell Wall-binding Domain Of ...mentioning

confidence: 99%

“…The endolysins of S. suis were shown to have more varieties in terms of its EAD and CBD than other streptococcal species. For example, PlySs2 contains only one EAD (CHAP) and one CBD (SH3_5), whereas PlySs9 contains two EADs, one amidase at the N terminus and one endopeptidase at the C terminus with a LysM CBD in the middle ( Vander Elst et al, 2020 ). PlySs2 showed a wider host range than PlySs9, which included MRSA, VISA, S. suis , Listeria , Staphylococcus simulans , Staphylococcus epidermidis , S. equi , S. agalactiae , S. pyogenes , S. sanguinis , GGS, GES, and S. pneumoniae .…”

Section: Enzymatically Active Domain and Cell Wall-binding Domain Of ...mentioning

confidence: 99%

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Endolysins against Streptococci as an antibiotic alternative

et al. 2022

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Multi-drug resistance has called for a race to uncover alternatives to existing antibiotics. Phage therapy is one of the explored alternatives, including the use of endolysins, which are phage-encoded peptidoglycan hydrolases responsible for bacterial lysis. Endolysins have been extensively researched in different fields, including medicine, food, and agricultural applications. While the target specificity of various endolysins varies greatly between species, this current review focuses specifically on streptococcal endolysins. Streptococcus spp. causes numerous infections, from the common strep throat to much more serious life-threatening infections such as pneumonia and meningitis. It is reported as a major crisis in various industries, causing systemic infections associated with high mortality and morbidity, as well as economic losses, especially in the agricultural industry. This review highlights the types of catalytic and cell wall-binding domains found in streptococcal endolysins and gives a comprehensive account of the lytic ability of both native and engineered streptococcal endolysins studied thus far, as well as its potential application across different industries. Finally, it gives an overview of the advantages and limitations of these enzyme-based antibiotics, which has caused the term enzybiotics to be conferred to it.

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“…Of the two, PlySs9 was reported more potent than PlySs2, with lower MIC and one log higher killing. 203 …”

Section: Introductionmentioning

confidence: 99%

How Good are Bacteriophages as an Alternative Therapy to Mitigate Biofilms of Nosocomial Infections

Singh¹,

Padmesh²,

Dwivedi³

et al. 2022

IDR

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Bacteria survive on any surface through the generation of biofilms that provide a protective environment to grow as well as making them drug resistant. Extracellular polymeric matrix is a crucial component in biofilm formation. The presence of biofilms consisting of common opportunistic and nosocomial, drug-resistant pathogens has been reported on medical devices like catheters and prosthetics, leading to many complications. Several approaches are under investigation to combat drug-resistant bacteria. Deployment of bacteriophages is one of the promising approaches to invade biofilm that may expose bacteria to the conditions adverse for their growth. Penetration into these biofilms and their destruction by bacteriophages is brought about due to their small size and ability of their progeny to diffuse through the bacterial cell wall. The other mechanisms employed by phages to infect biofilms may include their relocation through water channels to embedded host cells, replication at local sites followed by infection to the neighboring cells and production of depolymerizing enzymes to decompose viscous biofilm matrix, etc. Various research groups are investigating intricacies involved in phage therapy to mitigate the bacterial infection and biofilm formation. Thus, bacteriophages represent a good control over different biofilms and further understanding of phage-biofilm interaction at molecular level may overcome the clinical challenges in phage therapy. The present review summarizes the comprehensive details on dynamic interaction of phages with bacterial biofilms and the role of phage-derived enzymes – endolysin and depolymerases in extenuating biofilms of clinical and medical concern. The methodology employed was an extensive literature search, using several keywords in important scientific databases, such as Scopus, Web of Science, PubMed, ScienceDirect, etc. The keywords were also used with Boolean operator “And”. More than 250 relevant and recent articles were selected and reviewed to discuss the evidence-based data on the application of phage therapy with recent updates, and related potential challenges.

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Characterization of the Bacteriophage-Derived Endolysins PlySs2 and PlySs9 with In Vitro Lytic Activity against Bovine Mastitis Streptococcus uberis

Cited by 27 publications

References 42 publications

Substantiation of propitious “Enzybiotic” from two novel bacteriophages isolated from a wastewater treatment plant in Qatar

Substantiation of propitious “Enzybiotic” from two novel bacteriophages isolated from a wastewater treatment plant in Qatar

Endolysins against Streptococci as an antibiotic alternative

How Good are Bacteriophages as an Alternative Therapy to Mitigate Biofilms of Nosocomial Infections

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