Copyright © 2005 Elsevier B.V. All rights reserved.
Effects of lipid composition on the membrane activity and lipid phase behaviour of Vibrio sp. DSM14379 cells grown at various NaCl concentrations
Received 9 July 2004;
Abstract
The membrane lipid composition of living cells generally adjusts to the prevailing environmental and physiological conditions. In this study, membrane activity and lipid composition of the Gram-negative bacterium Vibrio sp. DSM14379, grown aerobically in a peptone-yeast extract medium supplemented with 0.5, 1.76, 3, 5 or 10% (w/v) NaCl, was determined. The ability of the membrane to reduce a spin label was studied by EPR spectroscopy under different salt concentrations in cell suspensions labeled with TEMPON. For lipid composition studies, cells were harvested in a late exponential phase and lipids were extracted with chloroform–methanol–water, 1:2:0.8 (v/v). The lipid polar head group and acyl chain compositions were determined by thin-layer and gas–liquid chromatographies. 31P-NMR spectroscopy was used to study the phase behaviour of the cell lipid extracts with 20 wt.% water contents in a temperature range from −10 to 50 °C. The results indicate that the ability of the membrane to reduce the spin label was highest at optimal salt concentrations. The composition of both polar head groups and acyl chains changed markedly with increasing salinity. The fractions of 16:0, 16:1 and 18:0 acyl chains increased while the fraction of 18:1 acyl chains decreased with increasing salinity. The phosphatidylethanolamine fraction correlated inversely with the lysophosphatidylethanolamine fraction, with phosphatidylethanolamine exhibiting a minimum, and lysophosphatidylethanolamine a maximum, at the optimum growth rate. The fraction of lysophosphatidylethanolamine was surprisingly high in the lipid extracts. This lipid can form normal micellar and hexagonal phases and it was found that all lipid extracts form a mixture of lamellar and normal isotropic liquid crystalline phases. This is an anomalous behaviour since the nonlamellar phases formed by total lipid extracts are generally of the reversed type.
Keywords: Lipid composition; Phase behaviour; Salt effect; Lysolipid; Vibrio sp.
Abbreviations: PG, phosphatidylglycerol; PE, phosphatidylethanolamine; lyso-PE, lysophosphatidylethanolamine; DPG, diphosphatidylglycerol; TLC, thin-layer chromatography; GLC, gas–liquid chromatography; 16:0, hexadecanoic acid; 18:0, octadecanoic acid; 14:1, cis-9-tetradecenoic acid; 16:1, cis-9-hexadecenoic acid; 18:1, cis-9-octadecenoic acid
Article Outline
- 1. Introduction
- 2. Materials and methods
- 2.1. Cell growth
- 2.2. EPR spectroscopy
- 2.3. Isolation and purification of lipids
- 2.4. Determination of polar head group composition
- 2.5. Determination of acyl chain composition
- 2.6. 31P-NMR spectroscopy
- 3. Results and discussion
- 3.1. Growth rates
- 3.2. Membrane spin label reduction process
- 3.3. Polar head group composition
- 3.4. Acyl chain composition
- 3.5. Phase equilibria in total membrane lipids
- Acknowledgements
- References






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