ArticleNucleic acids research2008
Promoter library designed for fine-tuned gene expression in Pichia pastoris.
Article in Nucleic acids research, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 112 papers.
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Who cites it
112 citing papers in PubMed, 311 citations in OpenAlex.
- Systematic optimization ofEngineering microbiology · 2026Review
- Engineering serine metabolism to enhance AOX1 promoter self-induction in formate dehydrogenase-deficient Komagataella phaffii.Microbial cell factories · 2026Article
- Dual transcriptional and post-transcriptional regulation by the bHLH proteins Rtg1 and RtgX in Komagataella phaffii.Nucleic acids research · 2026Article
- Adaptive Laboratory Evolution of Pichia pastoris by Serial Batch Cultivation.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Best Practices for Protein Expression in Pichia pastoris Combining Design of Experiments with an Automated Platform for Parallelized Cultivation in Small-Scale Stirred Tank Bioreactors.Methods in molecular biology (Clifton, N.J.) · 2026Article
- High-level expression of zearalenone lactonohydrolase ZenG in Pichia pastoris via combinatorial strategy.Archives of microbiology · 2025Article
- Next-generation stress-inducible Komagataella phaffii promoter variants.Microbial cell factories · 2025Article
- Promoter engineering for enhanced 3-hydroxypropionic acid production inSynthetic and systems biotechnology · 2025Article
- De novo promoter design method based on deep generative and dynamic evolution algorithm.Nucleic acids research · 2025Article
- Rational design and characterization of enhanced alcohol-inducible synthetic promoters inApplied and environmental microbiology · 2025Article
- Evaluation of the immune effect of foot-and-mouth disease virus-like particles derived fromFrontiers in microbiology · 2025Article
- Development of a bacterial gene transcription activating strategy based on transcriptional activator positive feedback.Journal of advanced research · 2024Article
- Combined strategies for improving the heterologous expression of a novel xylanase fromSynthetic and systems biotechnology · 2024Article
- Novel transcriptional regulation of the GAP promoter in Pichia pastoris towards high expression of heterologous proteins.Microbial cell factories · 2024Article
- Species-specific design of artificial promoters by transfer-learning based generative deep-learning model.Nucleic acids research · 2024Article
- An up-scaled biotechnological approach for phosphorus-depleted rye bran as animal feed.Bioresources and bioprocessing · 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
- Effect ofPeerJ · 2024Article
- Promoters in Pichia pastoris: A Toolbox for Fine-Tuned Gene Expression.Methods in molecular biology (Clifton, N.J.) · 2024Review
52 more citing papers are in PubMed but not listed here.
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Authors and funding
10 authors at 5 institutions in 3 countries.
Funding
Abstract
Although frequently used as protein production host, there is only a limited set of promoters available to drive the expression of recombinant proteins in Pichia pastoris. Fine-tuning of gene expression is often needed to maximize product yield and quality. However, for efficient knowledge-based engineering, a better understanding of promoter function is indispensable. Consequently, we created a promoter library by deletion and duplication of putative transcription factor-binding sites within the AOX1 promoter (P(AOX1)) sequence. This first library initially spanned an activity range between approximately 6% and >160% of the wild-type promoter activity. After characterization of the promoter library employing a green fluorescent protein (GFP) variant, the new regulatory toolbox was successfully utilized in a 'real case', i.e. the expression of industrial enzymes. Characterization of the library under repressing, derepressing and inducing conditions displayed at least 12 cis-acting elements involved in P(AOX1)-driven high-level expression. Based on this deletion analysis, novel short artificial promoter variants were constructed by combining cis-acting elements with basal promoter. In addition to improving yields and quality of heterologous protein production, the new P(AOX1) synthetic promoter library constitutes a basic toolbox to fine-tune gene expression in metabolic engineering and sequential induction of protein expression in synthetic biology.
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Registered trials
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.