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Applied and Environmental Microbiology, March 2002, p. 1088-1095, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1088-1095.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Effects of Pressure-Induced Membrane Phase Transitions on Inactivation of HorA, an ATP-Dependent Multidrug Resistance Transporter, in Lactobacillus plantarum

H. M. Ulmer,1 H. Herberhold,2 S. Fahsel,2 M. G. Gänzle,1* R. Winter,2 and R. F. Vogel1

Lehrstuhl für Technische Mikrobiologie, Weihenstephaner Steig 16, TU München, D-85350 Freising,1 FB Chemie, Physikalische Chemie I, Universität Dortmund, D-44221 Dortmund, Germany2

Received 9 July 2001/ Accepted 30 November 2001

The effects of pressure on cultures of Lactobacillus plantarum were characterized by determination of the viability and activity of HorA, an ATP-binding cassette multidrug resistance transporter. Changes in the membrane composition of L. plantarum induced by different growth temperatures were determined. Furthermore, the effect of the growth temperature of a culture on pressure inactivation at 200 MPa was determined. Cells were characterized by plate counts on selective and nonselective agar after pressure treatment, and HorA activity was measured by ethidium bromide efflux. Fourier transform-infrared spectroscopy and Laurdan fluorescence spectroscopy provided information about the thermodynamic phase state of the cytoplasmic membrane during pressure treatment. A pressure-temperature diagram for cell membranes was established. Cells grown at 37°C and pressure treated at 15°C lost >99% of HorA activity and viable cell counts within 36 and 120 min, respectively. The membranes of these cells were in the gel phase region at ambient pressure. In contrast, cells grown at 15°C and pressure treated at 37°C lost >99% of HorA activity and viable cell counts within 4 and 8 min, respectively. The membranes of these cells were in the liquid crystalline phase region at ambient pressure. The kinetic analysis of inactivation of L. plantarum provided further evidence that inactivation of HorA is a crucial step during pressure-induced cell death. Comparison of the biological findings and the membrane state during pressure treatment led to the conclusion that the inactivation of cells and membrane enzymes strongly depends on the thermodynamic properties of the membrane. Pressure treatment of cells with a liquid crystalline membrane at 0.1 MPa resulted in HorA inactivation and cell death more rapid than those of cells with a gel phase membrane at 0.1 MPa.

* Corresponding author. Mailing address: TU München, Lehrstuhl für Technische Mikrobiologie, Weihenstephaner Steig 16, 85350 Freising, Germany. Phone: 49 8161 713959. Fax: 49 8161 713327. E-mail: michael.gaenzle{at}bl.tum.de.

Applied and Environmental Microbiology, March 2002, p. 1088-1095, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1088-1095.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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