ArticleApplied and environmental microbiology2010
Secretion of an endogenous subtilisin by Pichia pastoris strains GS115 and KM71.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 29 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
- Molecular engineering of Komagataella phaffii for venom toxin production.Applied microbiology and biotechnology · 2026Review
- Characterizing the role of an endogenous serine protease KpSub2 in recombinant collagen degradation in Komagataella phaffii.Bioresources and bioprocessing · 2026Article
- Article
- Primary high-throughput screening of engineered phytases by online monitoring of the oxygen transfer rate of Komagataella phaffii.Microbial cell factories · 2025Article
- Recombinant Production of Food Allergens in Yeast Pichia pastoris.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Sequence Engineering of an Aspergillus niger Tannase to Produce in Pichia pastoris a Single-Chain Enzyme with High Specific Activity.Molecular biotechnology · 2022Article
- Comparison of the secretory murine DNase1 family members expressed in Pichia pastoris.PloS one · 2021Article
- AtPME17 is a functional Arabidopsis thaliana pectin methylesterase regulated by its PRO region that triggers PME activity in the resistance to Botrytis cinerea.Molecular plant pathology · 2020Article
- Holistic process development to mitigate proteolysis of a subunit rotavirus vaccine candidate produced in Pichia pastoris by means of an acid pH pulse during fed-batch fermentation.Biotechnology progress · 2020Article
- Comparative genome-scale analysis of Pichia pastoris variants informs selection of an optimal base strain.Biotechnology and bioengineering · 2020Article
- Bioproduction of Quercetin and Rutinose Catalyzed by Rutinosidase: Novel Concept of "Solid State Biocatalysis".International journal of molecular sciences · 2019Article
- Functional expression of trypsin from Streptomyces griseus by Pichia pastoris.Journal of industrial microbiology & biotechnology · 2012Article
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The methylotrophic yeast Pichia pastoris is widely used for the expression of heterologous enzymes. While the purity of the desired expression product is of major importance for many applications, we found that recombinant enzymes produced in methanol medium were contaminated by a 37-kDa endogenous yeast protease. This enzyme was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) but not by 1,10-phenanthroline, EDTA, and pepstatin A, suggesting the nature of a serine protease. Its secretion was abolished in P. pastoris strains GS115 and KM71 by specific mutagenesis of a subtilisin gene (SUB2) but not by inactivation of the gene encoding vacuolar proteinase B (PRB). Bioinformatic comparisons of Sub2 protein with subtilisins from other fungal genomes and phylogenetic analyses indicated that this enzyme is not an orthologue of the vacuolar protease cerevisin generally present in yeasts but is more closely related to another putative subtilisin found in a small number of yeast genomes. During growth of P. pastoris, Sub2 was produced as a secreted enzyme at a concentration of 10 microg/ml of culture supernatant after overexpression of the full-length SUB2 gene. During fermentative production of recombinant enzymes in methanol medium, 1 ml of P. pastoris culture supernatant was found to contain approximately 3 ng of Sub2, while the enzyme was not detected during growth in a medium containing glycerol as a carbon source. The mutant strain GS115-sub2 was subsequently used as a host for the production of recombinant proteases without endogenous subtilisin contamination.
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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.