Tailoring the Host Range of Ackermannviridae Bacteriophages through Chimeric Tailspike Proteins

Host range is a major determinant in the industrial utility of a bacteriophage. A model host range permits broad recognition across serovars of a target bacterium while avoiding cross-reactivity with commensal microbiota. Searching for a naturally occurring bacteriophage with ideal host ranges is ch...

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Bibliographic Details
Published in:Viruses 2023-01, Vol.15 (2), p.286
Main Authors: Gil, Jose, Paulson, John, Brown, Matthew, Zahn, Henriett, Nguyen, Minh M, Eisenberg, Marcia, Erickson, Stephen
Format: Article
Language:English
Subjects:
Ackermannviridae
bacteriophage
Bacteriophages - genetics
Caudovirales
Cross Reactions
Cross-reactivity
Design
E coli
Engineering
Food safety
Host range
Host Specificity
Industrial applications
Listeria
luciferase reporter phage
Pathogens
phage-based detection
Phages
Plasmids
Protein Engineering
Proteins
receptor-binding protein
Recombinant Fusion Proteins - metabolism
Salmonella
Salmonella enterica
Tailspike protein
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Summary:Host range is a major determinant in the industrial utility of a bacteriophage. A model host range permits broad recognition across serovars of a target bacterium while avoiding cross-reactivity with commensal microbiota. Searching for a naturally occurring bacteriophage with ideal host ranges is challenging, time-consuming, and restrictive. To address this, SPTD1.NL, a previously published luciferase reporter bacteriophage for , was used to investigate manipulation of host range through receptor-binding protein engineering. Similar to related members of the bacteriophage family, SPTD1.NL possessed a receptor-binding protein gene cluster encoding four tailspike proteins, TSP1-4. Investigation of the native gene cluster through chimeric proteins identified TSP3 as the tailspike protein responsible for detection. Further analysis of chimeric phages revealed that TSP2 contributed off-target recognition, whereas TSP1 and TSP4 were not essential for activity against any known host. To improve the host range of SPTD1.NL, TSP1 and TSP2 were sequentially replaced with chimeric receptor-binding proteins targeting . This engineered construct, called RBP-SPTD1-3, was a superior diagnostic reporter, sensitively detecting additional serovars while also demonstrating improved specificity. For industrial applications, bacteriophages of the family are thus uniquely versatile and may be engineered with multiple chimeric receptor-binding proteins to achieve a custom-tailored host range.
ISSN:1999-4915
1999-4915
DOI:10.3390/v15020286