Evaluating the effect of silver nanoparticles on bacteriophage lytic infection cycle-a mechanistic understanding
Bacteriophages and engineered nano-material (AgNPS) interactions is a relatively unexplored area of research. To answer the fundamental question whether bacteriophage lytic growth cycle is affected by the presence of AgNPs, laboratory experiments were performed with phages of Klebsiella pneumoniae, Delftia tsuruhatensis, Salmonella typhimurium, and Shigella flexneri using silver nanoparticles (AgNPs) with coating materials. One-step growth curves of bacteriophages indicated that the presence of these nanoparticles, and the associated ions of silver, produced pronounced effects on the lytic infection of certain bacteriophages. Effects included 96% reductions in post-infection phage yield in terms of plaque forming units (PFUs) after phages were incubated with silver nanoparticles and 28%–43% reductions from the presence of Ag+ alone. However, when Klebsiella pneumonia phage KL and Salmonella typhimurium phage Det7 were exposed to silver nanoparticles coated with poly-N-vinyl-2 pyrrolidone (PVP), an increase in final phage yield by as much as 250% was observed compared with the same phage not incubated with nanoparticles. A proposed mechanism, observed by transmission electron microscopy and verified using synthetic biology by which the nanoparticle binding phenotype can be produced, is that the binding of metal nanomaterial to phage virions results in potentially inhibitory effects. This binding was found to be dependent on the presence of exposed positively charged C-terminal amino-acid residues on the phage capsid surface, implied at first by amino-acid sequence comparisons between capsid proteins of the different phages used in this study. This was then proven experimentally using targeted DNA editing methods to fuse positive charged amino-acid residues to the coat protein C-terminus of non-binding phage. This induced the AgNP binding phenotype, as observed by TEM, DLS size measurements, and growth curve data that show the mutant constructs to be functionally inhibited after exposure to AgNPs. This research sets up a first platform for further research in the unexplored area of phage and AgNP interactions and provides useful findings.
Publication date: 15/08/2020
Author: Eddie Gilcrease, Ryan Williams, Ramesh Goel