ArticleFEMS yeast research2019
Engineering the expression system for Komagataella phaffii (Pichia pastoris): an attempt to develop a methanol-free expression system.
Article in FEMS yeast research, 2019. 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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13 citing papers in PubMed, 30 citations in OpenAlex.
- Advances in Key Genetic Elements and Strategies for High-Yield Heterologous Protein Expression inJournal of fungi (Basel, Switzerland) · 2026Review
- Systematic metabolic engineering of Komagataella phaffii for methanol-to-lactate bioconversion.Bioprocess and biosystems engineering · 2026Article
- Balancing Doses of EL222 and Light Improves Optogenetic Induction of Protein Production in Komagataella phaffii.Biotechnology and bioengineering · 2025Article
- Methanol-Free Protein Expression in Komagataella phaffii With Magnetic or Non-Magnetic Heating.Microbial biotechnology · 2025Article
- Enhanced recombinant xylanase B expression in Komagataella phaffii through metabolic engineering of methanol utilization.Archives of microbiology · 2025Article
- Novel transcriptional regulation of the GAP promoter in Pichia pastoris towards high expression of heterologous proteins.Microbial cell factories · 2024Article
- Review
- Enhanced Expression of Alcohol Dehydrogenase I inMicroorganisms · 2023Article
- Transcriptomic profiling of the yeast Komagataella phaffii in response to environmental alkalinization.Microbial cell factories · 2023Article
- Improving AOX1 promoter efficiency by overexpression of Mit1 transcription factor.Molecular biology reports · 2022Article
- Current advances ofFrontiers in microbiology · 2022Review
- Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris.Biotechnology for biofuels · 2021Article
- Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds.Antibiotics (Basel, Switzerland) · 2020Review
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Authors and funding
6 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
The construction of a methanol-free expression system of Komagataella phaffii (Pichia pastoris) was attempted by engineering a strong methanol-inducible DAS1 promoter using Citrobacter braakii phytase production as a model case. Constitutive expression of KpTRM1, formerly PRM1-a positive transcription regulator for methanol-utilization (MUT) genes of K. phaffii,was demonstrated to produce phytase without addition of methanol, especially when a DAS1 promoter was used but not an AOX1 promoter. Another positive regulator, Mxr1p, did not have the same effect on the DAS1 promoter, while it was more effective than KpTrmp1 on the AOX1 promoter. Removing a potential upstream repression sequence (URS) and multiplying UAS1DAS1 in the DAS1 promoter significantly enhanced the yield of C. braakii phytase with methanol-feeding, which surpassed the native AOX1 promoter by 80%. However, multiplying UAS1DAS1 did not affect the yield of methanol-free expression by constitutive KpTrm1p. Another important region to enhance the effect of KpTrm1p under a methanol-free condition was identified in the DAS1 promoter, and was termed ESPDAS1. Nevertheless, methanol-free phytase production using an engineered DAS1 promoter outperformed phytase production with the GAP promoter by 25%. Difference in regulation by known transcription factors on the AOX1 promoter and the DAS1 promoter was also illustrated.
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