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Strain-dependent selective antimicrobial action of cationic polyelectrolytes on Gram-negative bacteria

Rybkin, I.; Mirtic, J.; Majaron, H.; Hrescak, J.; Ceh, M.; Strancar, J.; Kristl, J.; Rijavec, T.; Lapanje, A.

2021-09-01 microbiology
10.1101/2021.08.31.458386 bioRxiv
Show abstract

Although electrostatic modification of bacterial surfaces using polyelectrolytes (PEs) is a convenient and versatile tool for biotechnological processes, the ambiguities in toxicity of PEs between various bacteria and the insufficient understanding of the mechanism of action of cationic PEs and their nano-thick shells formed around the bacteria create a bottleneck of the approach. Here, we show how the viability of two bacterial strains, Escherichia coli and Pseudomonas stutzeri, both from the Gram-negative group differs, when the cells are exposed to cationic PEs under different conditions. Although the cell wall architecture of the strains should be structurally similar, we found that the viability of E. coli was not affected by the electrostatic deposition of polyethyleneimine (PEI) or poly(allylamine) hydrochloride (PAH), whereas for P. stutzeri the deposition resulted in high death rates. The cells of E. coli proved to be suitable templates for Layer-by-Layer (LbL) modification, while in P. stutzeri a modified protocol with mild conditions had to be used to ensure the viability of the cells. Super resolution stimulated emission depletion (STED) microscopy allowed us to clearly visualize that after PE deposition onto the surface of the cells, the PEs could penetrate inside the cells of P. stutzeri, while forming a capsule around E. coli as expected. Therefore, this knowledge will help us select the most appropriate combinations of strains and PEs, for biotechnological processes or biomedical application, preventing unwanted toxicity.

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