ArticleNucleic acids research2020
Exchange of endogenous and heterogeneous yeast terminators in Pichia pastoris to tune mRNA stability and gene expression.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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28 citing papers in PubMed, 56 citations in OpenAlex.
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- Advancing recombinant protein expression in Komagataella phaffii: opportunities and challenges.FEMS yeast research · 2025Review
- Transforming non-conventional yeasts into key players in biotechnology: advances in synthetic biology applications.Frontiers in microbiology · 2025Review
- Regulatory significance of terminator: A systematic approach for dissecting terminator-mediated enhancement of upstream mRNA stability.Synthetic and systems biotechnology · 2025Article
- Designing strong inducible synthetic promoters in yeasts.Nature communications · 2024Article
- POMBOX: A Fission Yeast Cloning Toolkit for Molecular and Synthetic Biology.ACS synthetic biology · 2024Article
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- Genetic tools for metabolic engineering ofEngineering microbiology · 2023Review
- Functional characterization of the Komagataella phaffii 1033 gene promoter and transcriptional terminator.World journal of microbiology & biotechnology · 2023Article
- A roadmap to establish a comprehensive platform for sustainable manufacturing of natural products in yeast.Nature communications · 2023Review
- Evaluation of reference genes for transcript analyses in Komagataella phaffii (Pichia pastoris).Fungal biology and biotechnology · 2023Article
- Avoiding entry into intracellular protein degradation pathways by signal mutations increases protein secretion in Pichia pastoris.Microbial biotechnology · 2022Article
- Identification of genome integration sites for developing a CRISPR-based gene expression toolkit in Yarrowia lipolytica.Microbial biotechnology · 2022Article
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11 authors at 3 institutions in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
In the yeast Saccharomyces cerevisiae, terminator sequences not only terminate transcription but also affect expression levels of the protein-encoded upstream of the terminator. The non-conventional yeast Pichia pastoris (syn. Komagataella phaffii) has frequently been used as a platform for metabolic engineering but knowledge regarding P. pastoris terminators is limited. To explore terminator sequences available to tune protein expression levels in P. pastoris, we created a 'terminator catalog' by testing 72 sequences, including terminators from S. cerevisiae or P. pastoris and synthetic terminators. Altogether, we found that the terminators have a tunable range of 17-fold. We also found that S. cerevisiae terminator sequences maintain function when transferred to P. pastoris. Successful tuning of protein expression levels was shown not only for the reporter gene used to define the catalog but also using betaxanthin production as an example application in pathway flux regulation. Moreover, we found experimental evidence that protein expression levels result from mRNA abundance and in silico evidence that levels reflect the stability of mRNA 3'-UTR secondary structure. In combination with promoter selection, the novel terminator catalog constitutes a basic toolbox for tuning protein expression levels in metabolic engineering and synthetic biology in P. pastoris.
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