Journal
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
Volume 1838, Issue 7, Pages 1889-1897Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbamem.2014.03.012
Keywords
Pichia pastoris; Plasma membrane; Phospholipids; Fatty acid; Sterol; Sphingolipid
Categories
Funding
- Austrian Science Fund FWF [TRP 9]
- Federal Ministry of Science, Research and Economy (BMWFW)
- Federal Ministry of Traffic, Innovation and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol and ZIT - Technology Agency of the City of Vienna through the COMET
- Austrian Research Promotion Agency FFG
- Austrian Centre of Industrial Biotechnology (ACIB GmbH)
- Microbiology and Biochemistry program of the Goettingen Graduate School of Neurosciences and Molecular Biology (GGNB)
- Austrian Science Fund (FWF) [TRP 9] Funding Source: researchfish
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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. (C) 2014 Elsevier B.V. All rights reserved.
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