Wriessnegger Tamara, Gübitz Georg, Leitner Erich, Ingolic Elisabeth, Cregg James, de la Cruz Bernard J, Daum Günther
Institute of Biochemistry, Graz University of Technology, Petersgasse 12/2, Graz, Austria.
Biochim Biophys Acta. 2007 Apr;1771(4):455-61. doi: 10.1016/j.bbalip.2007.01.004. Epub 2007 Jan 12.
Highly purified peroxisomes from the yeast Pichia pastoris grown on methanol or oleic acid, respectively, were used to characterize the lipid composition of this organelle. For this purpose, an isolation procedure had to be adapted which yielded highly purified P. pastoris peroxisomes. When peroxisome proliferation was induced by growth on methanol, alcohol oxidase was the predominant peroxisomal protein. Cultivation of P. pastoris on oleic acid led to induction of a family of peroxisomal enzymes catalyzing fatty acid beta-oxidation, whose most prominent members were identified by mass spectrometry. On either carbon source, phosphatidylcholine and phosphatidylethanolamine were the major peroxisomal phospholipids, and cardiolipin was present in peroxisomal membranes at a substantial amount, indicating that this phospholipid is a true peroxisomal component. Ergosterol was the most abundant sterol of P. pastoris peroxisomal membranes irrespective of the culture conditions. The fatty acid composition of whole cells and peroxisomes was highly affected by cultivation of P. pastoris on oleic acid. Under these conditions, oleic acid became the predominant fatty acid in phospholipids from total cell and peroxisomal extracts. Thus, oleic acid was not only utilized as an appropriate carbon source but also as a building block for complex membrane lipids. In summary, our data provide first insight into biochemical properties of P. pastoris peroxisomal membranes, which may become important for the biotechnological use of this yeast.
分别在甲醇或油酸上生长的巴斯德毕赤酵母中高度纯化的过氧化物酶体，被用于表征该细胞器的脂质组成。为此，必须调整一种分离程序，以获得高度纯化的巴斯德毕赤酵母过氧化物酶体。当通过在甲醇上生长诱导过氧化物酶体增殖时，醇氧化酶是主要的过氧化物酶体蛋白。在油酸上培养巴斯德毕赤酵母会导致诱导出一系列催化脂肪酸β-氧化的过氧化物酶体酶，其最主要的成员通过质谱鉴定。在任何一种碳源上，磷脂酰胆碱和磷脂酰乙醇胺都是主要的过氧化物酶体磷脂，心磷脂大量存在于过氧化物酶体膜中，表明这种磷脂是真正的过氧化物酶体成分。无论培养条件如何，麦角固醇都是巴斯德毕赤酵母过氧化物酶体膜中最丰富的固醇。巴斯德毕赤酵母在油酸上培养对全细胞和过氧化物酶体的脂肪酸组成有很大影响。在这些条件下，油酸成为全细胞和过氧化物酶体提取物中磷脂的主要脂肪酸。因此，油酸不仅被用作合适的碳源，还被用作复杂膜脂的构建块。总之，我们的数据首次揭示了巴斯德毕赤酵母过氧化物酶体膜的生化特性，这可能对该酵母的生物技术应用具有重要意义。
