Rupcić J, Milin C, Marić V
Department of Chemistry and Biochemistry, Medical Faculty, University of Rijeka, Croatia.
Syst Appl Microbiol. 1999 Sep;22(3):486-91. doi: 10.1016/S0723-2020(99)80059-9.
Candida lipolytica yeast was grown batchwise on n-hexadecane as the carbon and energy source. Ceramides were quantitatively isolated from total lipids of exponential and stationary phase cells by a combination of column chromatography and preparative high-performance thin-layer chromatography. After acid methanolysis their composition was analyzed by gas-liquid chromatography. The ceramide content of the exponential phase cells was two times higher than the one of the stationary phase cells. The composition of long-chain base moiety of ceramides did not change significantly during the growth. In both growth phases 19-phytosphingosine was the major long-chain base. However, the fatty acid composition of ceramides changed greatly during the growth. In the exponential growth phase, ceramides contained predominantly fatty acids greater than 20 carbon atoms, while fatty acids shorter than 20 atoms predominated in ceramides of the stationary phase, 16:0 being the main one. In the exponential growth phase fatty acid moiety of ceramides was characterized by unusually high degree of unsaturation and relatively high proportion of odd-numbered fatty acids. However, the proportion of both, unsaturated and odd-numbered fatty acid decreased significantly in ceramides of the stationary phase. The unexpected finding was the absence of fatty acid hydroxylation of ceramides in the exponential phase cells and unusually low degree of hydroxylation in the stationary phase.
解脂耶氏酵母以正十六烷作为碳源和能源进行分批培养。通过柱色谱和制备型高效薄层色谱相结合的方法,从指数生长期和稳定期细胞的总脂质中定量分离出神经酰胺。经酸性甲醇解后,采用气液色谱法分析其组成。指数生长期细胞的神经酰胺含量是稳定期细胞的两倍。在生长过程中,神经酰胺长链碱基部分的组成没有显著变化。在两个生长阶段,19-植物鞘氨醇都是主要的长链碱基。然而,神经酰胺的脂肪酸组成在生长过程中发生了很大变化。在指数生长期,神经酰胺主要含有大于20个碳原子的脂肪酸,而在稳定期神经酰胺中,小于20个碳原子的脂肪酸占主导,其中16:0是主要脂肪酸。在指数生长期,神经酰胺的脂肪酸部分具有异常高的不饱和度和相对较高比例的奇数碳脂肪酸。然而,在稳定期神经酰胺中,不饱和脂肪酸和奇数碳脂肪酸的比例均显著下降。意外的发现是,指数生长期细胞的神经酰胺不存在脂肪酸羟基化现象,而稳定期的羟基化程度异常低。