ArticleApplied and environmental microbiology2011
GAP promoter library for fine-tuning of gene expression in Pichia pastoris.
Article in Applied and environmental microbiology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 61 papers.
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61 citing papers in PubMed, 164 citations in OpenAlex.
- Systematic optimization ofEngineering microbiology · 2026Review
- Transcriptional regulation: efficient genetic engineering tools for non-conventional yeasts.FEMS yeast research · 2026Review
- Concentration and Purification of Proteins Produced by Pichia pastoris by TFF with Ceramic Membranes.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Promoter engineering for enhanced 3-hydroxypropionic acid production inSynthetic and systems biotechnology · 2025Article
- Transcriptome analysis reveals methanol metabolism variations for the growth damage caused by overexpression of chimeric transactivators inSynthetic and systems biotechnology · 2025Article
- An optimised promoter and signal peptide improves methionine production of a genetically engineeredFrontiers in microbiology · 2025Article
- Enhancing D-lactic acid production by optimizing the expression of D-LDH gene in methylotrophic yeast Komagataella phaffii.Biotechnology for biofuels and bioproducts · 2024Article
- Saccharomyces cerevisiae whole cell biotransformation for the production of aldehyde flavors and fragrances.Applied microbiology and biotechnology · 2024Article
- Novel transcriptional regulation of the GAP promoter in Pichia pastoris towards high expression of heterologous proteins.Microbial cell factories · 2024Article
- Recent progress on heterologous protein production in methylotrophic yeast systems.World journal of microbiology & biotechnology · 2024Review
- Identification of a novel growth-associated promoter for biphasic expression of heterogenous proteins inApplied and environmental microbiology · 2024Article
- Impact of cell wall polysaccharide modifications on the performance of Pichia pastoris: novel mutants with enhanced fitness and functionality for bioproduction applications.Microbial cell factories · 2024Article
- Effect ofPeerJ · 2024Article
- Promoters in Pichia pastoris: A Toolbox for Fine-Tuned Gene Expression.Methods in molecular biology (Clifton, N.J.) · 2024Review
- Enhanced Expression of Alcohol Dehydrogenase I inMicroorganisms · 2023Article
- Genetic tools for metabolic engineering ofEngineering microbiology · 2023Review
- Systematic sequence engineering enhances the induction strength of the glucose-regulated GTH1 promoter of Komagataella phaffii.Nucleic acids research · 2023Article
- Biosynthesis of High-Active Hemoproteins by the Efficient Heme-Supply Pichia Pastoris Chassis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- Functional characterization of the Komagataella phaffii 1033 gene promoter and transcriptional terminator.World journal of microbiology & biotechnology · 2023Article
- The advent of plant cells in bioreactors.Frontiers in plant science · 2023Review
1 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A library of engineered promoters of various strengths is a useful genetic tool that enables the fine-tuning and precise control of gene expression across a continuum of broad expression levels. The methylotrophic yeast Pichia pastoris is a well-established expression host with a large academic and industrial user base. To facilitate manipulation of gene expression spanning a wide dynamic range in P. pastoris, we created a functional promoter library through mutagenesis of the constitutive GAP promoter. Using yeast-enhanced green fluorescent protein (yEGFP) as the reporter, 33 mutants were chosen to form the functional promoter library. The 33 mutants spanned an activity range between ∼0.6% and 19.6-fold of the wild-type promoter activity with an almost linear fluorescence intensity distribution. After an extensive characterization of the library, the broader applicability of the results obtained with the yEGFP reporter was confirmed using two additional reporters (β-galactosidase and methionine adenosyltransferase [MAT]) at the transcription and enzyme activity levels. Furthermore, the utility of the promoter library was tested by investigating the influence of heterologous MAT gene expression levels on cell growth and S-adenosylmethionine (SAM) production. The extensive characterization of the promoter strength enabled identification of the optimal MAT activity (around 1.05 U/mg of protein) to obtain maximal volumetric SAM production. The promoter library permits precise control of gene expression and quantitative assessment that correlates gene expression level with physiologic parameters. Thus, it is a useful toolbox for both basic and applied research in P. pastoris.
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