ArticleMicrobial cell factories2017
Using a vector pool containing variable-strength promoters to optimize protein production in Yarrowia lipolytica.
Article in Microbial cell factories, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers.
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52 citing papers in PubMed, 137 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
- Division of labour enables Yarrowia lipolytica synthetic consortia to achieve higher growth rates from starch.Microbial cell factories · 2026Review
- New Yarrowia lipolytica chassis strains for industrial enzyme production.Microbial cell factories · 2025Article
- Advances and perspectives in genetic expression and operation for the oleaginous yeastSynthetic and systems biotechnology · 2024Review
- Establishing Gene Expression and Knockout Methods in Esteya vermicola CBS115803.Molecular biotechnology · 2024Article
- Screening of broad-host expression promoters for shuttle expression vectors in non-conventional yeasts and bacteria.Microbial cell factories · 2024Article
- Customizable and stable multilocus chromosomal integration: a novel glucose-dependent selection system in Aureobasidium spp.Biotechnology for biofuels and bioproducts · 2024Article
- 'Mother(Nature) knows best' - hijacking nature-designed transcriptional programs for enhancing stress resistance and protein production in Yarrowia lipolytica; presentation of YaliFunTome database.Microbial cell factories · 2024Article
- Up Front Unfolded Protein Response Combined with Early Protein Secretion Pathway Engineering inInternational journal of molecular sciences · 2023Article
- Refactoring the architecture of a polyketide gene cluster enhances docosahexaenoic acid production in Yarrowia lipolytica through improved expression and genetic stability.Microbial cell factories · 2023Article
- Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production.Microbial cell factories · 2023Article
- Genome-wide analysis reveals Hsf1 maintains high transcript abundance of target genes controlled by strong constitutive promoter in Saccharomyces cerevisiae.Biotechnology for biofuels and bioproducts · 2023Article
- Transcriptome-based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae.Journal of microbiology (Seoul, Korea) · 2023Article
- Biosynthesis of Fatty Acid Derivatives by RecombinantJournal of fungi (Basel, Switzerland) · 2023Article
- Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system.FEMS yeast research · 2023Article
- Over-expression of α-bisabolene by metabolic engineering of Yarrowia lipolytica employing a golden gate DNA assembly toolbox.Biotechnology notes (Amsterdam, Netherlands) · 2023Article
- Employing Engineered Enolase Promoter for Efficient Expression ofInternational journal of molecular sciences · 2022Article
- Elevating Phospholipids ProductionInternational journal of molecular sciences · 2022Article
- Identification of genome integration sites for developing a CRISPR-based gene expression toolkit in Yarrowia lipolytica.Microbial biotechnology · 2022Article
- Enhancing very long chain fatty acids production in Yarrowia lipolytica.Microbial cell factories · 2022Article
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9 authors at 2 institutions in 1 country.
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Abstract
backgroundThe yeast Yarrowia lipolytica is an increasingly common biofactory. To enhance protein expression, several promoters have been developed, including the constitutive TEF promoter, the inducible POX2 promotor, and the hybrid hp4d promoter. Recently, new hp4d-inspired promoters have been created that couple various numbers of UAS1 tandem elements with the minimal LEU2 promoter or the TEF promoter. Three different protein-secretion signaling sequences can be used: preLip2, preXpr2, and preSuc2.
resultsTo our knowledge, our study is the first to use a set of vectors with promoters of variable strength to produce proteins of industrial interest. We used the more conventional TEF and hp4d promoters along with five new hybrid promoters: 2UAS1-pTEF, 3UAS1-pTEF, 4UAS1-pTEF, 8UAS1-pTEF, and hp8d. We compared the production of RedStar2, glucoamylase, and xylanase C when strains were grown on three media. As expected, levels of RedStar2 and glucoamylase were greatest in the strain with the 8UAS1-pTEF promoter, which was stronger. However, surprisingly, the 2UAS1-pTEF promoter was associated with the greatest xylanase C production and activity. This finding underscored that stronger promoters are not always better when it comes to protein production. We therefore developed a method for easily identifying the best promoter for a given protein of interest. In this gateway method, genes for YFP and α-amylase were transferred into a pool of vectors containing different promoters and gene expression was then analyzed. We observed that, in most cases, protein production and activity were correlated with promoter strength, although this pattern was protein dependent.
conclusionsProtein expression depends on more than just promoter strength. Indeed, promoter suitability appears to be protein dependent; in some cases, optimal expression and activity was obtained using a weaker promoter. We showed that using a vector pool containing promoters of variable strength can be a powerful tool for rapidly identifying the best producer for a given protein of interest.
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