ArticleJournal of industrial microbiology & biotechnology2013
Disruption of YPS1 and PEP4 genes reduces proteolytic degradation of secreted HSA/PTH in Pichia pastoris GS115.
Article in Journal of industrial microbiology & biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 66 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
- Evolutionary Adaptation and Targeted Metabolic Engineering Synergistically Improve Methanol-Derived Single-Cell Protein Production in Pichia pastoris.Biotechnology journal · 2026Article
- Characterizing the role of an endogenous serine protease KpSub2 in recombinant collagen degradation in Komagataella phaffii.Bioresources and bioprocessing · 2026Article
- Establishment and application of an RNAi system inFrontiers in bioengineering and biotechnology · 2025Article
- Fusion Partner Facilitates Expression of Cell-Penetrating Peptide L2 inAntibiotics (Basel, Switzerland) · 2024Article
- Applications of the Methylotrophic YeastJournal of fungi (Basel, Switzerland) · 2024Review
- Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions inJournal of fungi (Basel, Switzerland) · 2024Article
- Expression and Purification of Cp3GT: Structural Analysis and Modeling of a Key Plant Flavonol-3-O Glucosyltransferase fromBiotech (Basel (Switzerland)) · 2024Article
- Effect and mechanism of signal peptide and maltose on recombinant type III collagen production in Pichia pastoris.Applied microbiology and biotechnology · 2023Article
- Improving therapeutic protein secretion in the probiotic yeast Saccharomyces boulardii using a multifactorial engineering approach.Microbial cell factories · 2023Article
- A streamlined strain engineering workflow with genome-wide screening detects enhanced protein secretion in Komagataella phaffii.Communications biology · 2022Article
- Engineering Saccharomyces cerevisiae for the production and secretion of Affibody molecules.Microbial cell factories · 2022Article
- Unusual substrate specificity in GH family 12: structure-function analysis of glucanases Bgh12A and Xgh12B from Aspergillus cervinus, and Egh12 from Thielavia terrestris.Applied microbiology and biotechnology · 2022Article
- Enhancement of HSA-pFSHβ production by disruptingFrontiers in microbiology · 2022Article
- A versatile toolbox for CRISPR-based genome engineering in Pichia pastoris.Applied microbiology and biotechnology · 2021Article
- Global transcriptome profiling reveals genes responding to overproduction of a small secretory, a high cysteine- and a high glycosylation-bearing protein inBiotechnology reports (Amsterdam, Netherlands) · 2021Article
- Customized yeast cell factories for biopharmaceuticals: from cell engineering to process scale up.Microbial cell factories · 2021Review
- A Novel GH Family 20 β-N-acetylhexosaminidase With Both Chitosanase and Chitinase Activity FromFrontiers in molecular biosciences · 2021Article
- A multistrategy approach for improving the expression of E. coli phytase in Pichia pastoris.Journal of industrial microbiology & biotechnology · 2020Article
- Histoplasma capsulatum 100-kilodalton antigen: recombinant production, characterization, and evaluation of its possible application in the diagnosis of histoplasmosis.Applied microbiology and biotechnology · 2020Article
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Authors and funding
8 authors at 3 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Human serum albumin (HSA) and human parathyroid hormone (1-34) [PTH (1-34)] fusion protein [HSA/PTH (1-34)] is a promising long-acting form of PTH (1-34) for osteoporosis treatment. Secretory expression of intact HSA/PTH (1-34) in Pichia pastoris GS115 was accompanied by two degradation fragments, with molecular weights around 66 kDa, in addition to the well-known ~45 kDa HSA-truncated fragment, resulting in a low yield of intact protein. In this study, two internal cleavage sites were identified in the PTH (1-34) portion of the fusion protein by Western Blot analysis. To minimize proteolytic cleavages, several protease genes including PEP4 (encoding proteinase A), PRB1 (proteinase B) and seven YPSs genes (yapsin family members) were knocked out respectively by disruption of the individual genes and the selective combinations. Reduced degradation was observed by single disruption of either PEP4 gene or YPS1 gene, and the lowest level of degradation was observed in a pep4△yps1△ double disruptant. After 72 h of induction, more than 80 % of the HSA/PTH (1-34) secreted by the pep4△yps1△ double disruptant remained intact, in comparison to only 30 % with the wild-type strain.
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