ArticleNature protocols2009
Engineering complex-type N-glycosylation in Pichia pastoris using GlycoSwitch technology.
Article in Nature protocols, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 108 papers.
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Who cites it
108 citing papers in PubMed, 265 citations in OpenAlex.
- Yarrowia lipolytica and Komagataella phaffii (Pichia pastoris) as Hosts for Heterologous Expression of Lipases.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Systematic optimization ofEngineering microbiology · 2026Review
- Engineering dextranase for the directed preparation of low-molecular-weight dextran.Food chemistry. Molecular sciences · 2026Article
- Metabolic engineering ofApplied and environmental microbiology · 2026Review
- Dose-dependent structural and electron-density features in the lytic polysaccharide monooxygenase NcAA9D.Acta crystallographica. Section D, Structural biology · 2026Article
- Host-Informed Engineering of Glycosylation in Komagataella phaffii for Human Biantennary N-Linked Glycans.Research square · 2026Article
- A Brief History of the Pichia pastoris Expression System.Methods in molecular biology (Clifton, N.J.) · 2026Article
- The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.Frontiers in bioengineering and biotechnology · 2026Review
- Development of a Komagataella phaffii-based expression system for high-activity anti-SARS-CoV-2 neutralizing antibodies.AMB Express · 2025Article
- The Impact of Methanol Concentration on Recombinant Protein Glycosylation in Pichia pastoris SuperMan5.Microbial biotechnology · 2025Article
- Editor's Choice Protein engineering strategies to develop lectins by design.Glycobiology · 2025Review
- Illuminating Glucomannan Synthases To Explore Cell Wall Synthesis Bottlenecks.ACS synthetic biology · 2025Article
- A toolkit for facilitating markerless integration of expression cassettes in Komagataella phaffii via CRISPR/Cas9.Microbial cell factories · 2025Article
- Adaptation of Aglycosylated Monoclonal Antibodies for Improved Production in Komagataella phaffii.Biotechnology and bioengineering · 2025Article
- Structural and Functional Glycosylation of the Abdala COVID-19 Vaccine.Glycobiology · 2025Article
- Different results despite high homology: Comparative expression of human and murine DNase1 in Pichia pastoris.PloS one · 2025Article
- A comparative guide to expression systems for phage lysin production.Essays in biochemistry · 2024Review
- Backstage Heroes-Yeast in COVID-19 Research.International journal of molecular sciences · 2024Review
- Immobilized enzyme cascade for targeted glycosylation.Nature chemical biology · 2024Article
- Recent progress on heterologous protein production in methylotrophic yeast systems.World journal of microbiology & biotechnology · 2024Review
48 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Here we provide a protocol for engineering the N-glycosylation pathway of the yeast Pichia pastoris. The general strategy consists of the disruption of an endogenous glycosyltransferase gene (OCH1) and the stepwise introduction of heterologous glycosylation enzymes. Each engineering step results in the introduction of one glycosidase or glycosyltransferase activity into the Pichia endoplasmic reticulum or Golgi complex and consists of a number of stages: transformation with the appropriate GlycoSwitch vector, small-scale cultivation of a number of transformants, sugar analysis and heterologous protein expression analysis. If desired, the resulting clone can be further engineered by repeating the procedure with the next GlycoSwitch vector. Each engineering step takes approximately 3 weeks. The conversion of any wild-type Pichia strain into a strain that modifies its glycoproteins with Gal(2)GlcNAc(2)Man(3)GlcNAc(2)N-glycans requires the introduction of five GlycoSwitch vectors. Three examples of the full engineering procedure are provided to illustrate the results that can be expected.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.