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TitleGenetically manipulated phages with improved pH resistance for oral administration in veterinary medicine
Author(s)Nóbrega, Franklin Luzia
Costa, Ana Rita
Santos, J.
Siliakus, Melvin F.
van Lent, Jan W. M.
Kengen, Servé W. M.
Azeredo, Joana
Kluskens, Leon
Phage Biology
Issue date15-Dec-2016
PublisherNature Publishing Group
JournalScientific Reports
CitationNóbrega, Franklin L.; Costa, Ana Rita; Santos, J.; Siliakus, Melvin F.; van Lent, Jan W. M.; Kengen, Servé W. M.; Azeredo, Joana; Kluskens, Leon D., Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine. Scientific Reports, 6(39235), 1-12, 2016
Abstract(s)Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. Here we report a simple and efficient alternative, consisting in the genetic engineering of phages to display lipids on their surfaces. Escherichia coli phage T7 was used as a model and the E. coli PhoE signal peptide was genetically fused to its major capsid protein (10A), enabling phospholipid attachment to the phage capsid. The presence of phospholipids on the mutant phages was confirmed by High Performance Thin Layer Chromatography, Dynamic Light Scattering and phospholipase assays. The stability of phages was analysed in simulated GIT conditions, demonstrating improved stability of the mutant phages with survival rates 102107 pfu.mL1 higher than wild-type phages. Our work demonstrates that phage engineering can be a good strategy to improve phage tolerance to GIT conditions, having promising application for oral administration in veterinary medicine.
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Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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