title: The relationship between membrane damage, release of protein and loss of viability in Escherichia coli exposed to high hydrostatic pressure.
abstract: The aim of this work was to examine a possible association between resistance of two Escherichia coli strains to high hydrostatic pressure and the susceptibility of their cell membranes to pressure-induced damage. Cells were exposed to pressures between 100 and 700 MPa at room temperature (approximately 20 degrees C) in phosphate-buffered-saline. In the more pressure-sensitive strain E. coli 8164, loss of viability occurred at pressures between 100 MPa and 300 MPa and coincided with irreversible loss of membrane integrity as indicated by uptake of propidium iodide (PI) and leakage of protein of molecular mass between 9 and 78 kDa from the cells. Protein release increased to a maximum at 400 MPa then decreased, possibly due to intracellular aggregation at the higher pressures. In the pressure-resistant strain E. coli J1, PI was taken up during pressure treatment but not after decompression indicating that cells were able to reseal their membranes. Loss of viability in strain J1 coincided with the transient loss of membrane integrity between approximately 200 MPa and 600 MPa. In E. coli J1 leakage of protein occurred before loss of viability and the released protein was of low molecular mass, between 8 and 11 kDa and may have been of periplasmic origin. In these two strains differences in pressure resistance appeared to be related to differences in the ability of their membranes to withstand disruption by pressure. However it appears that transient loss of membrane integrity during pressure can lead to cell death irrespective of whether cells can reseal their membranes afterwards.
authors.affiliation: Department of Food and Nutrition Sciences, The University of Reading, PO Box 226, Whiteknights, Reading RG6 6AP, UK.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Klotz
authors.forename: Pilar
authors.initials: P
authors.lastname: Mañas
authors.forename: Bernard M
authors.initials: BM
authors.lastname: Mackey
mesh_terms: D002021:Buffers; D002463:Cell Membrane Permeability / Q000502:physiology*; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D006874:Hydrostatic Pressure; D050296:Microbial Viability; D008970:Molecular Weight; D011419:Propidium / Q000378:metabolism; D013194:Staining and Labeling; D013696:Temperature
chemical_list: D002021:Buffers; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D011419:Propidium