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An Endolysin LysSE24 by Bacteriophage LPSE1 Confers Specific Bactericidal Activity against Multidrug-Resistant Salmonella Strains.

ABSTRACT: Salmonella is responsible for a wide range of infections and is a constant threat to public health, particularly in light of emerging antibiotic resistance. The use of bacteriophages and phage endolysins as specific antibacterial agents is a promising strategy to control this bacterial infection. Endolysins are important proteins during the process of bacteria lysis by bacteriophages. In this study, we identify a novel endolysin, named LysSE24. LysSE24 was predicted to possess N-acetylmuramidases activity, with a molecular mass of ca. 17.4 kDa and pI 9.44. His-tagged LysSE24 was heterologously expressed and purified by Ni-NTA chromatography. LysSE24 exhibited optimal bactericidal activity against Salmonella Enteritidis ATCC 13076 at a concentration of 0.1 ?M. Salmonella population (measured by OD600 nm) decreased significantly (p < 0.05) after 10 min of incubation in combination with the outer membrane permeabilizer in vitro. It also showed antibacterial activity against a panel of 23 tested multidrug-resistant Salmonella strains. Bactericidal activity of LysSE24 was evaluated in terms of pH, temperature, and ionic strength. It was very stable with different pH (4.0 to 10.0) at different temperatures (20 to 60 °C). Both K+ and Na+ at concentrations between 0.1 to 100 mM showed no effects on its bactericidal activity, while a high concentration of Ca2+ and Mg2+ showed efficacy. Transmission electron microscopy revealed that exposure to 0.1 ?M LysSE24 for up to 5 min caused a remarkable modification of the cell shape of Salmonella Enteritidis ATCC 13076. These results indicate that recombinant LysSE24 represents a promising antimicrobial activity against Salmonella, especially several multidrug-resistant Salmonella strains. Further studies can be developed to improve its bactericidal activity without the need for pretreatment with outer membrane-destabilizing agents by synthetic biology methods.

SUBMITTER: Ding Y

PROVIDER: S-EPMC7284969 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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An Endolysin LysSE24 by Bacteriophage LPSE1 Confers Specific Bactericidal Activity against Multidrug-Resistant Salmonella Strains.

Ding Yifeng Y Zhang Yu Y Huang Chenxi C Wang Jia J Wang Xiaohong X

Microorganisms 20200515 5

Salmonella is responsible for a wide range of infections and is a constant threat to public health, particularly in light of emerging antibiotic resistance. The use of bacteriophages and phage endolysins as specific antibacterial agents is a promising strategy to control this bacterial infection. Endolysins are important proteins during the process of bacteria lysis by bacteriophages. In this study, we identify a novel endolysin, named LysSE24. LysSE24 was predicted to possess N-ac ...[more]

PMID: 32429030

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Project description:Lytic bacteriophages (or phages) drive bacterial mortality by elaborating exquisite abilities to bind, breach, and destroy bacterial cell membranes and subjugate critical bacterial cell functions. These antimicrobial activities make phages ideal candidates to serve as, or provide sources of, biological control measures for bacterial pathogens. In this study, we isolated the Myoviridae phage vB_BanS_Bcp1 (here referred to as Bcp1) from landfill soil, using a Bacillus anthracis host. The antimicrobial activities of both Bcp1 and its encoded endolysin, PlyB, were examined across different B. cereussensu lato group species, including B. cereussensu stricto, Bacillus thuringiensis, and Bacillus anthracis, with pathogenic potential in humans and multiple different uses in biotechnological applications. The Bcp1 phage infected only a subset (11 to 66%) of each B. cereussensu lato species group tested. In contrast, functional analysis of purified PlyB revealed a potent bacteriolytic activity against all B. cereussensu lato isolates tested (n = 79). PlyB was, furthermore, active across broad temperature, pH, and salt ranges, refractory to the development of resistance, bactericidal as a single agent, and synergistic with a second endolysin, PlyG. To confirm the potential for PlyB as an antimicrobial agent, we demonstrated the efficacy of a single intravenous treatment with PlyB alone or combination with PlyG in a murine model of lethal B. anthracis infection. Overall, our findings show exciting potential for the Bcp1 bacteriophage and the PlyB endolysin as potential new additions to the antimicrobial armamentarium.IMPORTANCE Organisms of the Bacillus cereussensu lato lineage are ubiquitous in the environment and are responsible for toxin-mediated infections ranging from severe food poisoning (B. cereussensu stricto) to anthrax (Bacillus anthracis). The increasing incidence of many of these infections, combined with the specter of antibiotic resistance, has created a need for novel antimicrobials with potent activity, including bacteriophages (or phages) and phage-encoded products (i.e., endolysins). In this study, we describe a broadly infective phage, Bcp1, and its encoded endolysin, PlyB, which exhibited a rapidly bacteriolytic effect against all B. cereussensu lato isolates tested with no evidence of evolving resistance. Importantly, PlyB was highly efficacious in a mouse model of lethal bacteremia with B. anthracis Both the Bcp1 phage and the PlyB endolysin represent novel mechanisms of action compared to antibiotics, with potential applications to address the evolving problem of antimicrobial resistance.

Project description:The continued rise and spread of antimicrobial resistance among bacterial pathogens pose a serious challenge to global health. Countering antimicrobial-resistant pathogens requires a multifaceted effort that includes the discovery of novel therapeutic approaches. Here, we establish the capacity of the human CXC chemokines CXCL9 and CXCL10 to kill multidrug-resistant Gram-negative bacteria, including New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae and colistin-resistant members of the family Enterobacteriaceae that harbor the mobile colistin resistance protein MCR-1 and thus possess phosphoethanolamine-modified lipid A. Colistin-resistant K. pneumoniae isolates affected by genetic mutation of the PmrA/PmrB two-component system, a chromosomally encoded regulator of lipopolysaccharide modification, and containing 4-amino-4-deoxy-l-arabinose-modified lipid A were also found to be susceptible to chemokine-mediated antimicrobial activity. However, loss of PhoP/PhoQ autoregulatory control, caused by disruption of the gene encoding the negative regulator MgrB, limited the bactericidal effects of CXCL9 and CXCL10 in a variable, strain-specific manner. Cumulatively, these findings provide mechanistic insight into chemokine-mediated antimicrobial activity, highlight disparities amongst determinants of colistin resistance, and suggest that chemokine-mediated bactericidal effects merit additional investigation as a therapeutic avenue for treating infections caused by multidrug-resistant pathogens.IMPORTANCE As bacterial pathogens become resistant to multiple antibiotics, the infections they cause become increasingly difficult to treat. Carbapenem antibiotics provide an essential clinical barrier against multidrug-resistant bacteria; however, the dissemination of bacterial enzymes capable of inactivating carbapenems threatens the utility of these important antibiotics. Compounding this concern is the global spread of bacteria invulnerable to colistin, a polymyxin antibiotic considered to be a last line of defense against carbapenem-resistant pathogens. As the effectiveness of existing antibiotics erodes, it is critical to develop innovative antimicrobial therapies. To this end, we demonstrate that the chemokines CXCL9 and CXCL10 kill the most concerning carbapenem- and colistin-resistant pathogens. Our findings provide a unique and timely foundation for therapeutic strategies capable of countering antibiotic-resistant "superbugs."

Dataset Information

An Endolysin LysSE24 by Bacteriophage LPSE1 Confers Specific Bactericidal Activity against Multidrug-Resistant Salmonella Strains.

Publications

An Endolysin LysSE24 by Bacteriophage LPSE1 Confers Specific Bactericidal Activity against Multidrug-Resistant <i>Salmonella</i> Strains.

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