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Applied and Environmental Microbiology, June 2006, p. 4232-4238, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.02904-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Energetics and Surface Properties of Pseudomonas putida DOT-T1E in a Two-Phase Fermentation System with 1-Decanol as Second Phase

Grit Neumann,1 Sjef Cornelissen,1,2 Frank van Breukelen,1,2 Steffi Hunger,3 Holger Lippold,4 Norbert Loffhagen,5 Lukas Y. Wick,5 and Hermann J. Heipieper1*

Department of Bioremediation, Centre for Environmental Research Leipzig-Halle (UFZ), Permoserstr. 15, 04318 Leipzig, Germany,1 Wageningen University and Research Centre, Costerweg 50, 6701 BH Wageningen, The Netherlands,2 Centre for Environmental Biotechnology (UBZ), Permoserstr. 15, 04318 Leipzig, Germany,3 Institute of Interdisciplinary Isotope Research (IIF), Permoserstr. 15, 04318 Leipzig, Germany,4 Department of Environmental Microbiology, Centre for Environmental Research Leipzig-Halle (UFZ), Permoserstr. 15, 04318 Leipzig, Germany5

Received 9 December 2005/ Accepted 2 April 2006

The solvent-tolerant strain Pseudomonas putida DOT-T1E was grown in batch fermentations in a 5-liter bioreactor in the presence and absence of 10% (vol/vol) of the organic solvent 1-decanol. The growth behavior and cellular energetics, such as the cellular ATP content and the energy charge, as well as the cell surface hydrophobicity and charge, were measured in cells growing in the presence and absence of 1-decanol. Although the cells growing in the presence of 1-decanol showed an about 10% reduced growth rate and a 48% reduced growth yield, no significant differences were measured either in the ATP and potassium contents or in the energy charge, indicating that the cells adapted completely at the levels of membrane permeability and energetics. Although the bacteria needed additional energy for adaptation to the presence of the solvent, they were able to maintain or activate electron transport phosphorylation, allowing homeostasis of the ATP level and energy charge in the presence of the solvent, at the price of a reduced growth yield. On the other hand, significantly enhanced cell hydrophobicities and more negative cell surface charges were observed in cells grown in the presence of 1-decanol. Both reactions occurred within about 10 min after the addition of the solvent and were significantly different after killing of the cells with toxic concentrations of HgCl2. This adaptation of the surface properties of the bacterium to the presence of solvents seems to be very similar to previously observed reactions on the level of lipopolysaccharides, with which bacteria adapt to environmental stresses, such as heat shock, antibiotics, or low oxygen content. The results give clear physiological indications that the process with P. putida DOT-T1E as the biocatalyst and 1-decanol as the solvent is a stable system for two-phase biotransformations that will allow the production of fine chemicals in economically sound amounts.

* Corresponding author. Mailing address: Department of Bioremediation, Centre for Environmental Research Leipzig-Halle (UFZ), Permoserstr. 15, 04318 Leipzig, Germany. Phone: 49 341 235 2772. Fax: 49 341 235 2492. E-mail: hermann.heipieper{at}ufz.de.

Applied and Environmental Microbiology, June 2006, p. 4232-4238, Vol. 72, No. 6
0099-2240/06/$08.00+0     doi:10.1128/AEM.02904-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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