Increasing temperatures of cultivation caused the cellular phospholipid content of C. glutamicum to decrease significantly. Simultaneously, the melting point of phospholipid fractions, i. e. the temperature of the ordered-disordered phase transition, increased. A hypothetical connection between the cellular phospholipid composition and the temperature-depended lysine efflux is discussed.Most bacteria alter their membrane lipid composition, and hence membrane fluidity, in response to changes in growth temperature. These alterations are often in the fatty acyl constituents of phospholipids and glycolipids and include changes in the saturation or length of fatty acyl chains in addition to variations in the proportion of cyclic or branched fatty acids (RUSSEL 1984). Normal cell function requires membrane lipid bilayers to be in a largely fluid state (MENDOZA and &ONAN 1983). However, a t low temperatures, membrane lipid bilayers undergo reversible order-disorder phase transitions as demonstrated by several methods (e. g. differential scanning calorimetry) (LEONARD et aZ. 1981). I n a recent paper concerning the influence of temperature on the lysine formation of C . glutarnicum, we reported on a decrease of the proportion of unsaturated fatty acids in the whole lipid fraction of this microorganism when grown a t increasing temperatures (HILLIGER et at. 1984). Furthermore, it was shown using freeze-fracture electron microscopy that, after quenching of the bacteria from temperatures below their growth temperature, the intramembraneous particles segregate and particle-free smooth areas of different sizes appear. Segregation started a t higher temperatures when C . glutarnicurn was grown a t higher temperatures (RICHTER et al. 1985). Such morphological pertubations may be induced by the order-disorder transitions of the paraffin chains within the hydrophobic core of the membrane (LEONARD et al. 1981). The aim of this work was to demonstrate directly such transitions of the cellular phospholipid fraction as a function of growth temperature by using differential scanning calorimetry (DSC).
Materials and methodsStrain and conditions of cultivation: Corynebaeterium glutamicum 9366 resistant to S-2-aminoethyl-L-cysteine and requiring homoserine was grown for 48 h in a mineral salt medium containing (g/l) : sucrose 75, (NH,),S04 30, KH,PO, 2, CaCO, 10, MgSO, -7 H,O 0.3, Peso,. 7 H,O 0.02, MnSO, 0.02, L-methionine 0.5, L-threonine 0.7, thiamine 0.01, biotin 0.0001, pH 7.0. Cultivations were carried out on rotary shakers (180 r.p.m., 5 om-strokes) in 500-ml flasks containing 50 ml medium at various temperatures as indicated.Extraction of lipids and thin-layer chromatography : Extraction of lipids and thin-layer chromatography were performed as described previously (GRHFE et al. 1982). Diphosphatidylglycerol (DPG) and phosphatidylglycerol (PG) were identified through parallel application of commercial samples of the two phospholipids (KOCH-LIGHT Ltd., England). Lipid phosphorus was estimated according to HESSE and GELLER (1965)....