Impedimetric analysis on the mass transfer properties of intact and competent E. coli cells

Biochim Biophys Acta Biomembr. 2019 Jan;1861(1):9-16. doi: 10.1016/j.bbamem.2018.10.010. Epub 2018 Oct 17.

Abstract

Competent Escherichia coli cells are commonly used in bacterial transformation owing to its high permeability for bioorganic macromolecules like plasmid DNA. However, the mass transfer property of competent E. coli cell has not fully investigated. In the present study, mass transfer coefficients of competent and intact E. coli cells in deionized water were evaluated by impedimetric analysis of the release of cytoplasmic compounds. Because competent cells have a higher permeability after chemical treatment, the lumped mass transfer coefficient of a competent cell was approximately 6.5 times larger than that of an intact cell at room temperature. Release of cytoplasmic components was accelerated at an elevated temperature of 42 °C, which is the heat shock temperature used during bacterial transformation. At this elevated temperature, assessed lumped mass transfer coefficients of intact and competent E. coli cells were 9.28 × 10-4 min-1 and 97.10 × 10-4 min-1, respectively. Significant increase in the mass transfer coefficient of the competent cell is caused by cytolysis of cells. The double layer capacitances were also assessed from the electrochemical spectra confirming the enhanced ion release from E. coli cells and rupture of the competent cell under prolonged exposure at the elevated temperature. Impedimetric detection of the ion release with analyses using an equivalent circuit model provides a method to evaluate mass transfer properties of biomolecules.

Keywords: Competence; Impedance; Mass transfer; Membrane; Osmotic pressure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biochemistry / methods
  • Calcium Chloride / chemistry
  • Cell Membrane / chemistry
  • Electric Impedance*
  • Electrochemistry / methods*
  • Escherichia coli / cytology*
  • Escherichia coli / physiology
  • Glycerol / chemistry
  • Osmosis
  • Plasmids / genetics
  • Spectrophotometry
  • Temperature

Substances

  • Calcium Chloride
  • Glycerol