Characterization and Protein Analysis of Escherichia coli Resistant to a Quaternary Ammonium Compound

  • MAEDA TAKUYA
    Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima
  • TANAKA SHINGO
    Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima
  • NAGAMUNE HIDEAKI
    Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima
  • KOURAI HIROKI
    Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima

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A resistant strain of Escherichia coil IFO12713, resistant against the quaternary ammonium compound (QAC), N-dodecylpyridinium iodide (P-12), was developed by using a standard broth dilution method and physiologically characterized by comparison to a sensitive strain. The cell surface hydrophobicity cultivated in the resistance-acquiring process gradually decreased and the lipopolysaccharides in the outer membrane increased. This result indicated that the cell surface of the resistant strain was more hydrophobic than that of the sensitive strain. One of the resistant strains isolated from the process also had resistance to other antibiotics and organic solvents. Rapid ethidium efflux was observed in the resistant strain compared to the sensitive strain. To elucidate the mechanism of resistance on a molecular level, protein analysis was performed with 2D-PAGE for the entire cellular fraction, and SDS-PAGE for the outer or inner membrane fractions. It was seen that 18 spots and bands of proteins specifically increased in the resistant strain. From the physiological characterization and the protein analysis, it was suggested that the QAC-resistant strain acquired its resistance by responding to antimicrobial stresses, stabilizing the outer membrane to control the adsorption/binding reaction between the cell and agent, and activating the efflux pump against the antimicrobial agents.

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