Danevcic Tjasa, Rilfors Leif, Strancar Janez, Lindblom Göran, Stopar David
University of Ljubljana, Biotechnical Faculty, Biological Centre, Department of Food Technology, Laboratory of Microbiology, Slovenia.
Biochim Biophys Acta. 2005 Jun 15;1712(1):1-8. doi: 10.1016/j.bbamem.2005.03.013. Epub 2005 Apr 15.
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. (31)P-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 degrees 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.
活细胞的膜脂质组成通常会根据当前的环境和生理条件进行调整。在本研究中,测定了革兰氏阴性菌嗜盐弧菌DSM14379在补充有0.5%、1.76%、3%、5%或10%(w/v)NaCl的蛋白胨 - 酵母提取物培养基中好氧生长时的膜活性和脂质组成。通过电子顺磁共振光谱(EPR)研究了在不同盐浓度下,用TEMPON标记的细胞悬液中膜还原自旋标记物的能力。对于脂质组成研究,在指数生长后期收获细胞,并用氯仿 - 甲醇 - 水(1:2:0.8,v/v)提取脂质。通过薄层色谱和气相 - 液相色谱测定脂质极性头部基团和酰基链组成。使用(31)P - NMR光谱研究了在 - 10至50摄氏度温度范围内含水量为20 wt.%的细胞脂质提取物的相行为。结果表明,在最佳盐浓度下,膜还原自旋标记物的能力最高。极性头部基团和酰基链的组成都随着盐度的增加而显著变化。随着盐度增加,16:0、16:1和18:0酰基链的比例增加,而18:1酰基链的比例下降。磷脂酰乙醇胺部分与溶血磷脂酰乙醇胺部分呈负相关关系,在最佳生长速率时,磷脂酰乙醇胺呈现最小值,溶血磷脂酰乙醇胺呈现最大值。在脂质提取物中,溶血磷脂酰乙醇胺的比例出奇地高。这种脂质可以形成正常的胶束相和六方相,并且发现所有脂质提取物都形成层状相和正常各向同性液晶相的混合物。这是一种异常行为,因为总脂质提取物形成的非层状相通常是反向类型的。