Abstract

Previous studies from our lab have shown that the antimicrobial peptide F1 obtained from the milk fermentation by Lactobacillus paracasei FX-6 derived from Tibetan kefir was different from common antimicrobial peptides; specifically, F1 simultaneously inhibited the growth of Gram-negative and Gram-positive bacteria. Here, we present follow-on work demonstrating that after the antimicrobial peptide F1 acts on either Escherichia coli ATCC 25922 (E. coli) or Staphylococcus aureus ATCC 63589 (S. aureus), their respective bacterial membranes were severely deformed. This deformation allowed leakage of potassium and magnesium ions from the bacterial membrane. The interaction between the antimicrobial peptide F1 and the bacterial membrane was further explored by artificially simulating the bacterial phospholipid membranes and then extracting them. The study results indicated that after the antimicrobial peptide F1 interacted with the bacterial membranes caused significant calcein leakage that had been simulated by different liposomes. Furthermore, transmission electron microscopy observations revealed that the phospholipid membrane structure was destroyed and the liposomes presented aggregation and precipitation. Quartz Crystal Microbalance with Dissipation (QCM-D) results showed that the antimicrobial peptide F1 significantly reduced the quality of liposome membrane and increased their viscoelasticity. Based on the study's findings, the phospholipid membrane particle size was significantly increased, indicating that the antimicrobial peptide F1 had a direct effect on the phospholipid membrane. Conclusively, the antimicrobial peptide F1 destroyed the membrane structure of both Gram-negative and Gram-positive bacteria by destroying the shared components of their respective phospholipid membranes which resulted in leakage of cell contents and subsequently cell death.

Highlights

  • The phospholipid bilayer is the foundation of all bacterial cell membranes and antimicrobial peptides usually target this membrane to kill bacteria [1, 2]

  • The cell morphology is the typical morphology of S. aureus with a uniform cytoplasm, clear and complete cell wall and cell membrane, and smooth surface

  • After 1 h incubation with the antimicrobial peptide F1, the S. aureus cell membrane was almost mixed with the entire cell and the surface contour became blurred

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Summary

Introduction

The phospholipid bilayer is the foundation of all bacterial cell membranes and antimicrobial peptides usually target this membrane to kill bacteria [1, 2]. Quartz Crystal Microbalance with Dissipation (QCM-D) monitors the frequency (F) and energy dissipation (D) of the quartz crystal surface in real time, thereby monitoring the dynamics of the membrane solid support membrane surface, and measuring the quality and viscoelastic changes in the phospholipid membrane [11, 12] Due to their different sound penetration depths, higher octave frequencies monitor the information on the surface of the sensor, while lower octave frequencies are closer to the water surface. QCM-D is used to monitor the quality and viscoelastic changes at different depths of the phospholipid membrane on the surface of the quartz crystal in real time This information allows one to preliminarily infer the types of interactions between the antimicrobial peptide molecules and the phospholipid membrane [13, 14]

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