Protective effect of calcium on inactivation of Escherichia coli by high hydrostatic pressure.

The effect of divalent cations on the inactivation of Escherichia coli by high hydrostatic pressure was investigated. The presence of 0.5 mmol l-1 of CaCl2, MgCl2, MnCl2 and FeCl2 reduced pressure inactivation of E. coli MG1655, while 0.5 mmol l-1 of ZnCl2, NiCl2, CuCl2 and CoCl2 increased inactivation. Baroprotection by Ca2+ was found to be dose-dependent up to at least 80 mmol l-1 and was studied in more detail in terms of inactivation kinetics. Logarithmic survivor plots against time deviated from first order kinetics, suggesting that MG1655 cultures were heterogeneous with regard to pressure resistance. All cultures were shown to contain a small proportion of cells that were only slowly inactivated. Addition of Ca2+ increased the proportion of these tolerant cells in the cultures up to 1000-fold at 80 mmol l-1, but did not affect their inactivation rate. The addition of EDTA resulted in the opposite effect, lowering the proportion of pressure-tolerant cells in the cultures. Three pressure-resistant mutants of E. coli MG1655 were found to be more resistant to EDTA under pressure compared with MG1655, and were unaffected by Ca2+ under pressure. In addition, these mutants had a 30-40% lower Ca2+ content than MG1655. Based on these results, it is postulated that pressure killing of E. coli MG1655 is mediated primarily by the destabilization of Ca(2+)-binding components, and that the mutations underlying pressure resistance have resulted in pressure-stable targets with reduced Ca(2+)-binding affinity.

High-pressure inactivation and sublethal injury of pressure-resistant Escherichia coli mutants in fruit juices.

The potential of high-pressure-resistant mutants of Escherichia coli to survive high-pressure pasteurization in fruit juices and in low-pH buffers was investigated. Treatments with up to 500 MPa of pressure caused only a limited direct inactivation of the mutants but resulted in an accelerated low-pH inactivation during subsequent storage.

Escherichia coli mutants resistant to inactivation by high hydrostatic pressure.

Alternating cycles of exposure to high pressure and outgrowth of surviving populations were used to select for highly pressure-resistant mutants of Escherichia coli MG1655. Three barotolerant mutants (LMM1010, LMM1020, and LMM1030) were isolated independently by using outgrowth temperatures of 30, 37, and 42 degrees C, respectively. Survival of these mutants after pressure treatment for 15 min at ambient temperature was 40 to 85% at 220 MPa and 0.5 to 1.5% at 800 MPa, while survival of the parent strain, MG1655, decreased from 15% at 220 MPa to 2 x 10(-8)% at 700 MPa. Heat resistance of mutants LMM1020 and LMM1030 was also altered, as evident by higher D values at 58 and 60 degrees C and reduced z values compared to those for the parent strain. D and z values for mutant LMM1010 were not significantly different from those for the parent strain. Pressure sensitivity of the mutants increased from 10 to 50 degrees C, as opposed to the parent strain, which showed a minimum around 40 degrees C. The ability of the mutants to grow at moderately elevated pressure (50 MPa) was reduced at temperatures above 37 degrees C, indicating that resistance to pressure inactivation is unrelated to barotolerant growth. The development of high levels of barotolerance as demonstrated in this work should cause concern about the safety of high-pressure food processing.