Investigating the role of ERAD on antibody processing in glycoengineered<i>Saccharomyces cerevisiae</i>

Piirainen, Mari A.; Frey, Alexander D.

doi:10.1093/femsyr/foaa002

Cited by 7 publications

(4 citation statements)

References 53 publications

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“…Moreover, antibody clearance in an ALG3 deletion strain with functional ERAD was comparable to the clearance in wild-type strain indicating that an exposed α1-6 linked mannose is sufficient for ER clearance ( de Ruijter and Frey, 2015 ). Furthermore, deletion of HRD1 , but not of HTM1 or YOS9 reduced clearance rates in an ALG3 ALG11 deletion strain ( Piirainen and Frey, 2020 ). This indicates that glycan-independent mechanisms additionally can contribute to IgG clearance in yeast.…”

Section: Discussionmentioning

confidence: 99%

The Impact of Glycoengineering on the Endoplasmic Reticulum Quality Control System in Yeasts

Piirainen

Frey²

2022

Front. Mol. Biosci.

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Yeasts are widely used and established production hosts for biopharmaceuticals. Despite of tremendous advances on creating human-type N-glycosylation, N-glycosylated biopharmaceuticals manufactured with yeasts are missing on the market. The N-linked glycans fulfill several purposes. They are essential for the properties of the final protein product for example modulating half-lives or interactions with cellular components. Still, while the protein is being formed in the endoplasmic reticulum, specific glycan intermediates play crucial roles in the folding of or disposal of proteins which failed to fold. Despite of this intricate interplay between glycan intermediates and the cellular machinery, many of the glycoengineering approaches are based on modifications of the N-glycan processing steps in the endoplasmic reticulum (ER). These N-glycans deviate from the canonical structures required for interactions with the lectins of the ER quality control system. In this review we provide a concise overview on the N-glycan biosynthesis, glycan-dependent protein folding and quality control systems and the wide array glycoengineering approaches. Furthermore, we discuss how the current glycoengineering approaches partially or fully by-pass glycan-dependent protein folding mechanisms or create structures that mimic the glycan epitope required for ER associated protein degradation.

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Section: Discussionmentioning

confidence: 99%

The Impact of Glycoengineering on the Endoplasmic Reticulum Quality Control System in Yeasts

Piirainen

Frey²

2022

Front. Mol. Biosci.

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“…Aza et al [89] introduced N-glycosylation into laccase derived from Pleurotus eryngii, which improved its expression and activity in S. cerevisiae [89]. Glycosylation sites can also be added to increase protein secretion [115]. For instance, the secretion of keratinase was increased 5-and 1.8-fold by introducing glycosylation sites in the N-terminal and C-terminal regions, respectively [116].…”

Section: Glycosylation Pathway Engineeringmentioning

confidence: 99%

Engineering strategies for enhanced heterologous protein production by Saccharomyces cerevisiae

Zhao,

Ma,

Zhang

et al. 2024

Microb Cell Fact

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Microbial proteins are promising substitutes for animal- and plant-based proteins. S. cerevisiae, a generally recognized as safe (GRAS) microorganism, has been frequently employed to generate heterologous proteins. However, constructing a universal yeast chassis for efficient protein production is still a challenge due to the varying properties of different proteins. With progress in synthetic biology, a multitude of molecular biology tools and metabolic engineering strategies have been employed to alleviate these issues. This review first analyses the advantages of protein production by S. cerevisiae. The most recent advances in improving heterologous protein yield are summarized and discussed in terms of protein hyperexpression systems, protein secretion engineering, glycosylation pathway engineering and systems metabolic engineering. Furthermore, the prospects for efficient and sustainable heterologous protein production by S. cerevisiae are also provided.

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“… 2012 ) and ER-associated degradation (ERAD; Idiris et al . 2010 , Piirainen and Frey 2020 ). The most frequently used genetic engineering strategies are deletion and overexpression of related constituent genes.…”

Section: Introductionmentioning

confidence: 98%

CRISPR/Cas9-mediated point mutations improve α-amylase secretion in Saccharomyces cerevisiae

Wang

Chen

et al. 2022

FEMS Yeast Research

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The rapid expansion of the application of pharmaceutical proteins and industrial enzymes requires robust microbial workhorses for high protein production. The budding yeast Saccharomyces cerevisiae is an attractive cell factory due to its ability to perform eukaryotic post-translational modifications and to secrete proteins. Many strategies have been used to engineer yeast platform strains for higher protein secretion capacity. Herein, we investigated a line of strains that have previously been selected after UV random mutagenesis for improved α-amylase secretion. A total of 42 amino acid altering point mutations identified in this strain line were re-introduced into the parental strain AAC to study their individual effects on protein secretion. These point mutations included missense mutations (amino acid substitution), nonsense mutations (stop codon generation) and frameshift mutations. For comparison, single gene deletions for the corresponding target genes were also performed in this study. Eleven point mutations and seven gene deletions were found to effectively improve α-amylase secretion. These targets were involved in several bioprocesses, including cellular stresses, protein degradation, transportation, mRNA processing and export, DNA replication and repair, which indicates that the improved protein secretion capacity in the evolved strains is the result of the interaction of multiple intracellular processes. Our findings will contribute to the construction of novel cell factories for recombinant protein secretion.

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Investigating the role of ERAD on antibody processing in glycoengineeredSaccharomyces cerevisiae

Cited by 7 publications

References 53 publications

The Impact of Glycoengineering on the Endoplasmic Reticulum Quality Control System in Yeasts

The Impact of Glycoengineering on the Endoplasmic Reticulum Quality Control System in Yeasts

Engineering strategies for enhanced heterologous protein production by Saccharomyces cerevisiae

CRISPR/Cas9-mediated point mutations improve α-amylase secretion in Saccharomyces cerevisiae

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