title: Isolation and characterization of the plasma membrane from the yeast Pichia pastoris.
abstract: 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.
authors.affiliation: Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Karlheinz
authors.initials: K
authors.lastname: Grillitsch
authors.affiliation: Department for Plant Biochemistry, Albrecht-von-Haller Institute for Plant Sciences, Georg-August-University, Goettingen, Germany.
authors.forename: Pablo
authors.initials: P
authors.lastname: Tarazona
authors.affiliation: Institute of Biochemistry, Graz University of Technology, Austria.
authors.forename: Lisa
authors.initials: L
authors.lastname: Klug
authors.affiliation: Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Tamara
authors.initials: T
authors.lastname: Wriessnegger
authors.affiliation: Institute of Plant Sciences, University of Graz, Austria.
authors.forename: Günther
authors.initials: G
authors.lastname: Zellnig
authors.affiliation: Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Austria.
authors.forename: Erich
authors.initials: E
authors.lastname: Leitner
authors.affiliation: Department for Plant Biochemistry, Albrecht-von-Haller Institute for Plant Sciences, Georg-August-University, Goettingen, Germany.
authors.forename: Ivo
authors.initials: I
authors.lastname: Feussner
authors.affiliation: Austrian Centre of Industrial Biotechnology, Graz, Austria; Institute of Biochemistry, Graz University of Technology, Austria. Electronic address: guenther.daum@tugraz.at.
authors.forename: Günther
authors.initials: G
authors.lastname: Daum
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D004875:Ergosterol / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D010843:Pichia / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism; D013261:Sterols / Q000378:metabolism
chemical_list: D005227:Fatty Acids; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D013107:Sphingolipids; D013261:Sterols; D004875:Ergosterol
keywords: Fatty acid; Phospholipids; Pichia pastoris; Plasma membrane; Sphingolipid; Sterol