Improving recombinant eukaryotic membrane protein yields in Pichia pastoris:the importance of codon optimization and clone selection

Abstract

In the last 15 years, 80% of all recombinant proteins reported in the literature were produced in the bacterium, Escherichia coli, or the yeast, Pichia pastoris. Nonetheless, developing effective general strategies for producing recombinant eukaryotic membrane proteins in these organisms remains a particular challenge. Using a validated screening procedure together with accurate yield quantitation, we therefore wished to establish the critical steps contributing to high yields of recombinant eukaryotic membrane protein in P. pastoris. Whilst the use of fusion partners to generate chimeric constructs and directed mutagenesis have previously been shown to be effective in bacterial hosts, we conclude that this approach is not transferable to yeast. Rather, codon optimization and the preparation and selection of high-yielding P. pastoris clones are effective strategies for maximizing yields of human aquaporins.

Publication DOI: https://doi.org/10.3109/09687688.2011.602219
Divisions: College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
College of Health & Life Sciences > School of Biosciences
Uncontrolled Keywords: aquaporins,codon,electroporation,humans,lithium chloride,pichia,recombinant proteins,genetic transformation,Cell Biology,Molecular Biology
Full Text Link: http://informah ... 688.2011.602219
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2011-09
Authors: Öberg, Fredrik
Sjöhamn, Jennie
Conner, Matthew T.
Bill, Roslyn M. (ORCID Profile 0000-0003-1331-0852)
Hedfalk, Kristina

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