Adaptation of Escherichia Coli Spheroplasts to the Characterization of Antimicrobial Peptides

摘要: With activity against a broad range of microorganisms and low susceptibility to the resistance mechanisms developed by multidrug resistant organisms, naturally-occurring antimicrobial peptides (AMPs) have potential serve as effective agents combat ever-increasing number antibiotic-resistant illnesses. In addition various spectroscopic measurements, assays, molecular modeling, action novel AMPs can be further characterized using confocal microscopy. This imaging technique utilizes fluorescently-labeled or fluorophore-containing peptide conjugates examine localization in bacterial cells, providing insight into peptide's mechanism action. However, small size rod shape bacteria make it difficult acquire clear conclusive images determine cell. The present study focuses on adaptation Escherichia coli spheroplasts, which are spherical at least four times larger than normal bacteria, model for characterization AMPs. spheroplasts increases ease focusing target allows clearer with increased resolution more convincing evidence localization. equivalent taken from any angle sample. During optimization process this approach, we found that presence stabilizing agent, divalent magnesium cation, affects translocation cell-penetrating Ongoing work is focused finding suitable replacement spheroplast stabilization. successful E. protocols microscopy will allow system used wide variety molecules, including other cell-penetrating, non-antimicrobial peptides.