ArticlePloS one2011
Native-state stability determines the extent of degradation relative to secretion of protein variants from Pichia pastoris.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 52 citations in OpenAlex.
- Recombination and Incomplete Lineage Sorting Resolve the Enigma of Lysozyme Evolution.Journal of molecular evolution · 2026Article
- Different results despite high homology: Comparative expression of human and murine DNase1 in Pichia pastoris.PloS one · 2025Article
- Biophysical evolution of the receptor-binding domains of SARS-CoVs.Biophysical journal · 2023Article
- Pathway engineering facilitates efficient protein expression in Pichia pastoris.Applied microbiology and biotechnology · 2022Review
- Comparative transcriptomic analysis-based identification of the regulation of foreign proteins with different stabilities expressed inFrontiers in microbiology · 2022Article
- Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris.Biotechnology for biofuels · 2021Article
- Engineering of the unfolded protein response pathway in Pichia pastoris: enhancing production of secreted recombinant proteins.Applied microbiology and biotechnology · 2021Review
- Development of a dual specific growth rate-based fed-batch process for production of recombinant human granulocyte colony-stimulating factor in Pichia pastoris.Bioprocess and biosystems engineering · 2021Article
- A multistrategy approach for improving the expression of E. coli phytase in Pichia pastoris.Journal of industrial microbiology & biotechnology · 2020Article
- Identification of an Exceptionally Long Intron in themSphere · 2018Article
- Efficient Heterologous Production ofInternational journal of molecular sciences · 2018Article
- Fate of the UPR marker protein Kar2/Bip and autophagic processes in fed-batch cultures of secretory insulin precursor producing Pichia pastoris.Microbial cell factories · 2018Article
- Recycling of a selectable marker with a self-excisable plasmid in Pichia pastoris.Scientific reports · 2017Article
- Effects of UPR and ERAD pathway on the prolyl endopeptidase production in Pichia pastoris by controlling of nitrogen source.Journal of industrial microbiology & biotechnology · 2017Article
- Rapid screening of cellular stress responses in recombinant Pichia pastoris strains using metabolite profiling.Journal of industrial microbiology & biotechnology · 2017Article
- Nanofibrillar hydrogel scaffolds from recombinant protein-based polymers with integrin- and proteoglycan-binding domains.Journal of biomedical materials research. Part A · 2016Article
- Combined strategies for improving expression of Citrobacter amalonaticus phytase in Pichia pastoris.BMC biotechnology · 2015Article
- Pichia pastoris secretes recombinant proteins less efficiently than Chinese hamster ovary cells but allows higher space-time yields for less complex proteins.Biotechnology journal · 2014Article
- Decrease of UPR- and ERAD-related proteins in Pichia pastoris during methanol-induced secretory insulin precursor production in controlled fed-batch cultures.Microbial cell factories · 2014Article
- Biomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2014Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
We have investigated the relationship between the stability and secreted yield of a series of mutational variants of human lysozyme (HuL) in Pichia pastoris. We show that genes directly involved in the unfolded protein response (UPR), ER-associated degradation (ERAD) and ER-phagy are transcriptionally up-regulated more quickly and to higher levels in response to expression of more highly-destabilised HuL variants and those variants are secreted to lower yield. We also show that the less stable variants are retained within the cell and may also be targeted for degradation. To explore the relationship between stability and secretion further, two different single-chain-variable-fragment (scFv) antibodies were also expressed in P. pastoris, but only one of the scFvs gave rise to secreted protein. The non-secreted scFv was detected within the cell and the UPR indicators were pronounced, as they were for the poorly-secreted HuL variants. The non-secreted scFv was modified by changing either the framework regions or the linker to improve the predicted stability of the scFv and secretion was then achieved and the levels of UPR indicators were lowered Our data support the hypothesis that less stable proteins are targeted for degradation over secretion and that this accounts for the decrease in the yields observed. We discuss the secretion of proteins in relation to lysozyme amyloidosis, in particular, and optimised protein secretion, in general.
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