Projects per year
Abstract
N-glycosylation is an important posttranslational modification affecting the properties and quality of therapeutic proteins. Glycoengineering in yeast aims to produce proteins carrying human-compatible glycosylation, enabling the production of therapeutic proteins in yeasts. In this work, we demonstrate further development and characterization of a glycoengineering strategy in a Saccharomyces cerevisiae Δalg3 Δalg11 strain where a truncated Man3GlcNAc2 glycan precursor is formed due to a disrupted lipid-linked oligosaccharide synthesis pathway. We produced galactosylated complex-type and hybrid-like N-glycans by expressing a human galactosyltransferase fusion protein both with and without a UDP-glucose 4-epimerase domain from Schizosaccharomyces pombe. Our results showed that the presence of the UDP-glucose 4-epimerase domain was beneficial for the production of digalactosylated complex-type glycans also when extracellular galactose was supplied, suggesting that the positive impact of the UDP-glucose 4-epimerase domain on the galactosylation process can be linked to other processes than its catalytic activity. Moreover, optimization of the expression of human GlcNAc transferases I and II and supplementation of glucosamine in the growth medium increased the formation of galactosylated complex-type glycans. Additionally, we provide further characterization of the interfering mannosylation taking place in the glycoengineered yeast strain.
Original language | English |
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Pages (from-to) | 301-315 |
Number of pages | 15 |
Journal | Applied Microbiology and Biotechnology |
Volume | 106 |
Issue number | 1 |
Early online date | 15 Dec 2021 |
DOIs | |
Publication status | Published - Jan 2022 |
MoE publication type | A1 Journal article-refereed |
Keywords
- Galactosyltransferase
- Glucosamine
- Glycoengineering
- UDP-glucose 4-epimerase
- Yeast
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Dive into the research topics of 'Production of galactosylated complex-type N-glycans in glycoengineered Saccharomyces cerevisiae'. Together they form a unique fingerprint.Projects
- 1 Finished
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Mapping and tailoring the protein folding compartment for antibody expression in yeast
Frey, A. (Principal investigator), Salminen, H. (Project Member) & Niemelä, L. (Project Member)
01/09/2016 → 31/08/2020
Project: Academy of Finland: Other research funding
Equipment
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Bioeconomy Research Infrastructure
Seppälä, J. (Manager)
School of Chemical EngineeringFacility/equipment: Facility