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The humanization of N-glycosylation pathways in yeast

Nature Reviews Microbiology volume 3, pages 119–128 (2005)Cite this article

Abstract

Yeast and other fungal protein-expression hosts have been extensively used to produce industrial enzymes, and are often the expression system of choice when manufacturing costs are of primary concern. However, for the production of therapeutic glycoproteins intended for use in humans, yeast have been less useful owing to their inability to modify proteins with human glycosylation structures. Yeast N-glycosylation is of the high-mannose type, which confers a short half-life in vivo and thereby compromises the efficacy of most therapeutic glycoproteins. Several approaches to humanizing yeast N-glycosylation pathways have been attempted over the past decade with limited success. Recently however, advances in the glycoengineering of yeast and the expression of therapeutic glycoproteins with humanized N-glycosylation structures have shown significant promise — this review summarizes the most important developments in the field.

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Figure 1: Major N-glycosylation pathways in humans and yeast.
Figure 2: Structure of glycosylated glucocerebrosidase.
Figure 3: Electron micrographs of Pichia pastoris.
Figure 4: A working model for the cellular distribution of glycosyltransferases throughout the secretory pathway.
Figure 5: Type II membrane proteins.

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Acknowledgements

The authors are grateful to the outstanding scientists that provided the basic knowledge upon which the engineering of humanized yeast strains was made possible. In particular, we wish to express our gratitude to P. Robbins, R. Bretthauer, K. Moremen, C. Barlowe, M. Aebi and R. Hitzman for helpful discussions and support. Most importantly, we are grateful to the exceptional group of scientists we have been able to work with over the past four years. It is their work that has advanced the field in such a significant way and may lead to a paradigmatic change in the production of complex protein-based therapeutics.

Author information

Authors and Affiliations

  1. GlycoFi Inc., 21 Lafayette Street, Lebanon, 03766, New Hampshire, USA

    Stefan Wildt

  2. Department of Biological Sciences, and Department of Chemistry, Thayer School of Engineering, Dartmouth College, Hanover, 03755, New Hampshire, USA

    Tilllman U. Gerngross

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  1. Stefan Wildt
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Correspondence to Tilllman U. Gerngross.

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Related links

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DATABASES

Entrez

AOX1

Campylobacter jejuni

Erd2p

Escherichia coli

Kluyveromyces lactis

mnn1

Saccharomyces cerevisiae

SwissProt

Alg3p

Anp1p

Gls1p

Hoc1p

Mnn9p

Mnn10p

Mnn11p

Mns1p

Och1p

Sec12p

Van1p

FURTHER INFORMATION

Tillman Gerngross's laboratory

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Wildt, S., Gerngross, T. The humanization of N-glycosylation pathways in yeast. Nat Rev Microbiol 3, 119–128 (2005). https://doi.org/10.1038/nrmicro1087

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