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Glycosylation Profiling of Heterologous Proteins

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Pichia Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 103))

Abstract

The methylotrophic yeast Pichia pastoris has been widely exploited for its high-level expression of heterologous proteins by recombination of gene sequences of interest with the methanol-inducible alcohol oxidase gene (AOX1) promoter (1–3). Secreted and cytoplasmic expression of heterologous proteins at levels equivalent to Escherichia coli and significantly higher than in Saccharomyces cerevisiae has been achieved (4). In addition, P. pastoris cultures can be easily scaled up to high cell densities, and as a result, yields are also high on a volumetric basis (e.g., 12 g/L, for tetanus toxin fragment C [5], 2.5 g/L for invertase [6], 2.5 g/L for α-amylase [7]). Secreted products can comprise more than 80% of the protein in the medium (6). However, secretion is a complex process that is not only dependent on gene dosage, but also on other factors, such as signal sequence recognition and processing, proteolysis, and glycosylation.

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References

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Author information

Authors and Affiliations

  1. Center for Genetic Engineering and Biotechnology, Havana, Cuba

    José A. Cremata, Raquel Montesino, Omar Quintero & Rossana García

Authors
  1. José A. Cremata
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  2. Raquel Montesino
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  3. Omar Quintero
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  4. Rossana García
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Editor information

Editors and Affiliations

  1. Chiron Technologies/Center for Gene Therapy, San Diego, CA, USA

    David R. Higgins

  2. Oregon Graduate Institute of Science and Technology, Portland, OR, USA

    James M. Cregg

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© 1998 Humana Press Inc., Totowa, NJ

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Cremata, J.A., Montesino, R., Quintero, O., García, R. (1998). Glycosylation Profiling of Heterologous Proteins. In: Higgins, D.R., Cregg, J.M. (eds) Pichia Protocols. Methods in Molecular Biology, vol 103. Humana, Totowa, NJ. https://doi.org/10.1385/0-89603-421-6:95

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  • DOI: https://doi.org/10.1385/0-89603-421-6:95

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-0-89603-421-1

  • Online ISBN: 978-1-59259-578-5

  • eBook Packages: Springer Protocols

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