Grillitsch Karlheinz, Tarazona Pablo, Klug Lisa, Wriessnegger Tamara, Zellnig Günther, Leitner Erich, Feussner Ivo, Daum Günther
Austrian Centre of Industrial Biotechnology, Graz, Austria.
Department for Plant Biochemistry, Albrecht-von-Haller Institute for Plant Sciences, Georg-August-University, Goettingen, Germany.
Biochim Biophys Acta. 2014 Jul;1838(7):1889-97. doi: 10.1016/j.bbamem.2014.03.012. Epub 2014 Mar 26.
Despite similarities of cellular membranes in all eukaryotes, every compartment displays characteristic and often unique features which are important for the functions of the specific organelles. In the present study, we biochemically characterized the plasma membrane of the methylotrophic yeast Pichia pastoris with emphasis on the lipids which form the matrix of this compartment. Prerequisite for this effort was the design of a standardized and reliable isolation protocol of the plasma membrane at high purity. Analysis of isolated plasma membrane samples from P. pastoris revealed an increase of phosphatidylserine and a decrease of phosphatidylcholine compared to bulk membranes. The amount of saturated fatty acids in the plasma membrane was higher than in total cell extracts. Ergosterol, the final product of the yeast sterol biosynthetic pathway, was found to be enriched in plasma membrane fractions, although markedly lower than in Saccharomyces cerevisiae. A further characteristic feature of the plasma membrane from P. pastoris was the enrichment of inositol phosphorylceramides over neutral sphingolipids, which accumulated in internal membranes. The detailed analysis of the P. pastoris plasma membrane is discussed in the light of cell biological features of this microorganism especially as a microbial cell factory for heterologous protein production.
尽管所有真核生物的细胞膜具有相似性，但每个区室都具有独特的特征，这些特征对于特定细胞器的功能至关重要。在本研究中，我们对甲基营养型酵母毕赤酵母的质膜进行了生化特性分析，重点关注构成该区室基质的脂质。这项工作的前提是设计一种标准化且可靠的高纯度质膜分离方案。对毕赤酵母分离的质膜样品分析表明，与整体膜相比，磷脂酰丝氨酸增加，磷脂酰胆碱减少。质膜中饱和脂肪酸的含量高于总细胞提取物。麦角固醇是酵母固醇生物合成途径的终产物，虽然明显低于酿酒酵母，但在质膜组分中含量丰富。毕赤酵母质膜的另一个特征是肌醇磷酸神经酰胺相对于中性鞘脂的富集，后者在内膜中积累。本文结合该微生物的细胞生物学特征，特别是作为异源蛋白生产的微生物细胞工厂，对毕赤酵母质膜进行了详细分析。
