Bacteriophage endolysins: applications for food safety

Schmelcher, Mathias; Loessner, Martin J.

doi:10.1016/j.copbio.2015.10.005

Cited by 168 publications

(119 citation statements)

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“…Endolysins have a wide array of usage such as antimicrobial agents [1], food safety [2], against phytopathogenic bacteria [3], enzybiotics [4], disinfectants [5] etc. Endolysins have been categorized into four classes: i) glycosidases (muramidase), ii) endopeptidases, iii) amidohydrolases (amidase), and iv) lytic transglycosylases.…”

Section: Introductionmentioning

confidence: 99%

A computational assessment of pH-dependent differential interaction of T7 lysozyme with T7 RNA polymerase

Borkotoky

Murali

2017

BMC Struct Biol

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BackgroundT7 lysozyme (T7L), also known as N-acetylmuramoyl-L-alanine amidase, is a T7 bacteriophage gene product. It involves two functions: It can cut amide bonds in the bacterial cell wall and interacts with T7 RNA polymerase (T7RNAP) as a part of transcription inhibition. In this study, with the help of molecular dynamics (MD) calculations and computational interaction studies, we investigated the effect of varying pH conditions on conformational flexibilities of T7L and their influence on T7RNAP -T7L interactions.ResultsFrom the MD studies of the T7L at three different pH strengths viz. 5, neutral and 7.9 it was observed that T7L structure at pH 5 exhibited less stable nature with more residue level fluctuations, decrease of secondary structural elements and less compactness as compared to its counterparts: neutral pH and pH 7.9. The T-pad analysis of the MD trajectories identified local fluctuations in few residues that influenced the conformational differences in three pH strengths. From the docking of the minimum energy representative structures of T7L at different pH strengths (obtained from the free energy landscape analysis) with T7RNAP structures at same pH strengths, we saw strong interaction patterns at pH 7.9 and pH 5. The MD analysis of these complexes also confirmed the observations of docking study. From the combined in silico studies, it was observed that there are conformational changes in N-terminal and near helix 1 of T7L at different pH strengths, which are involved in the T7RNAP interaction, thereby varying the interaction pattern.ConclusionSince T7L has been used for developing novel therapeutics and T7RNAP one of the most biologically useful protein in both in-vitro and in vivo experiments, this in silico study of pH dependent conformational differences in T7L and the differential interaction with T7RNAP at different pH can provide a significant insight into the structural investigations on T7L and T7RNAP in varying pH environments.Electronic supplementary materialThe online version of this article (doi:10.1186/s12900-017-0077-9) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

A computational assessment of pH-dependent differential interaction of T7 lysozyme with T7 RNA polymerase

Borkotoky

Murali

2017

BMC Struct Biol

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“…Besides, endolysins are considered safe, not likely to generate bacterial resistance, and promising anti‐biofilm agents (Coffey et al . ; Schmelcher and Loessner ). For all these reasons, the application of purified phage endolysins against pathogenic bacteria has attracted considerable interest over the last decade.…”

Section: Methods Of Phage Detection: Advantages and Disadvantagesmentioning

confidence: 99%

“…This natural process is called lysis ‘from within’, but in the case of most Gram‐positive phages, their isolated endolysins are capable of lysing bacterial cells ‘from without’, with no phage infection required. These enzymes normally possess cell wall‐binding domains, involved in an extremely specific recognition of distinctive molecules, and enzymatically active domains, actually responsible of cell wall degradation (Loessner ; Schmelcher and Loessner ). This modular structure allows domain shuffling approaches to obtain chimeric molecules with desired specificity and improved activity (Fig.…”

Section: Methods Of Phage Detection: Advantages and Disadvantagesmentioning

confidence: 99%

Bacteriophages on dairy foods

2018

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This review focuses on the impact of bacteriophages on the manufacture of dairy foods. Firstly, the impact of phages of lactic acid bacteria in the dairy industry, where they are considered enemies, is discussed. The sources of phage contamination in dairy plants are detailed, with special emphasis on the rise of phage infections related to the growing use of cheese whey as ingredient. Other topics include traditional and new methods of phage detection, quantification and monitoring, and strategies of phage control in dairy plants, either of physical, chemical or biological nature. Finally, the use of phages or purified phage enzymes as allies to control pathogenic bacteria in the food industry is reviewed.

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“…The endolysin encoded by phage T5 is a well characterized endopeptidase that hydrolyzes the bond between L-alanine and D-glutamic acid of the peptidoglycan in E. coli (12). While native endolysins have been successfully applied as antimicrobials against Gram-positive bacteria including Staphylococcus aureus and streptococci, the outer membrane of Gram-negative bacteria generally prevents access to the peptidoglycan layer (13). To enable killing of Gram-negative pathogens using endolysins, a number of engineering strategies have been proposed.…”

Section: Introductionmentioning

confidence: 99%

Innolysins: A novel approach to engineer endolysins to kill Gram-negative bacteria

Zampara¹,

Sørensen²,

Grimon

et al. 2018

Preprint

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13Bacteriophage-encoded endolysins degrading the essential peptidoglycan of bacteria are promising 14 alternative antimicrobials to handle the global threat of antibiotic resistant bacteria. However, 15 endolysins have limited use against Gram-negative bacteria, since their outer membrane prevents 16 access to the peptidoglycan. Here we present Innolysins, a novel concept for engineering endolysins 17 that allows the enzymes to pass through the outer membrane, hydrolyse the peptidoglycan and kill 18 the target bacterium. Innolysins combine the enzymatic activity of endolysins with the binding 19 capacity of phage receptor binding proteins (RBPs). As our proof of concept, we used phage T5 20 endolysin and receptor binding protein Pb5, which binds irreversibly to the phage receptor FhuA 21 involved in ferrichrome transport in Escherichia coli. In total, we constructed twelve Innolysins 22 fusing endolysin with Pb5 or the binding domain of Pb5 with or without flexible linkers in between. 23While the majority of the Innolysins maintained their muralytic activity, Innolysin#6 also showed 24 bactericidal activity against E. coli reducing the number of bacteria by 1 log, thus overcoming the 25 outer membrane barrier. Using an E. coli fhuA deletion mutant, we demonstrated that FhuA is 26 required for bactericidal activity, supporting that the specific binding of Pb5 to its receptor on E. 27 coli is needed for the endolysin to access the peptidoglycan. Accordingly, Innolysin#6 was able to 28 kill other bacterial species that carry conserved FhuA homologs such as Shigella sonnei and 29 Pseudomonas aeruginosa. In summary, the Innolysin approach expands recent protein engineering 30 strategies allowing customization of endolysins by exploiting phage RBPs to specifically target 31Gram-negative bacteria. 32 IMPORTANCE 33The extensive use of antibiotics has led to the emergence of antimicrobial resistant bacteria 34 responsible for infections causing more than 50,000 deaths per year across Europe and the US. In 35 response, the World Health Organization has stressed an urgent need to discover new antimicrobials 36 to control in particular Gram-negative bacterial pathogens, due to their extensive multi-drug 37 resistance. However, the outer membrane of Gram-negative bacteria limits the access of many 38 antibacterial agents to their targets. Here, we developed a new approach, Innolysins that enable 39 endolysins to overcome the outer membrane by exploiting the binding specificity of phage receptor 40 binding proteins. As proof of concept, we constructed Innolysins against E. coli using the endolysin 41 and the receptor binding protein of phage T5. Given the rich diversity of phage receptor binding 42 proteins and their different binding specificities, our proof of concept paves the route for creating an 43 arsenal of pathogen specific alternative antimicrobials. 44 intracellular targets (1). Bacteriophages (phages), viruses that infect bacteria, have naturally 48 evolved mechanisms to overcome the outer membrane to infect their ...

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Bacteriophage endolysins: applications for food safety

Cited by 168 publications

References 103 publications

A computational assessment of pH-dependent differential interaction of T7 lysozyme with T7 RNA polymerase

A computational assessment of pH-dependent differential interaction of T7 lysozyme with T7 RNA polymerase

Bacteriophages on dairy foods

Innolysins: A novel approach to engineer endolysins to kill Gram-negative bacteria

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