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Membrane Protein Production in the Yeast P. pastoris

Part of the Methods in Molecular Biology book series (MIMB,volume 2507)


The first crystal structures of recombinant mammalian membrane proteins were solved using high-quality protein that had been produced in yeast cells. One of these, the rat Kv1.2 voltage-gated potassium channel, was synthesized in Pichia pastoris. Since then, this yeast species has remained a consistently popular choice of host for synthesizing eukaryotic membrane proteins because it is quick, easy, and cheap to culture and is capable of posttranslational modification. Very recent structures of recombinant membrane proteins produced in P. pastoris include a series of X-ray crystallography structures of the human vitamin K epoxide reductase and a cryo-electron microscopy structure of the TMEM206 proton-activated chloride channel from pufferfish. P. pastoris has also been used to structurally and functionally characterize a range of membrane proteins including tetraspanins, aquaporins, and G protein-coupled receptors. This chapter provides an overview of the methodological approaches underpinning these successes.

Key words

  • Recombinant membrane protein
  • Yeast
  • Komagataella pastoris
  • Methanol
  • AOX1

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  • DOI: 10.1007/978-1-0716-2368-8_10
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Buffered glycerol-complex medium


Buffered methanol-complex medium


Basal salts medium


Dissolved oxygen


High-performance liquid chromatography


Volume by volume


Weight by volume


Yeast extract peptone dextrose medium


Yeast extract peptone glycerol medium


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We acknowledge funding from the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G., A.J.R., and R.M.B.

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Correspondence to Roslyn M. Bill .

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Ayub, H. et al. (2022). Membrane Protein Production in the Yeast P. pastoris. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 2507. Humana, New York, NY.

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