ArticleMicrobial cell factories2016
Developing a set of strong intronic promoters for robust metabolic engineering in oleaginous Rhodotorula (Rhodosporidium) yeast species.
Article in Microbial cell factories, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 65 citations in OpenAlex.
- EngineeringSynthetic and systems biotechnology · 2026Article
- Enzyme-constrained genome-scale model of Yarrowia lipolytica predicts growth-phase specific metabolic engineering targets.Applied microbiology and biotechnology · 2026Article
- In silico characterization of Rhodotorula toruloides ELO-like elongases and production of very-long-chain fatty acids by expressing Rtelo2, RtKCR, RtHCD, and RtECR through IRES-mediated bicistrons.World journal of microbiology & biotechnology · 2024Article
- Microbial Squalene: A Sustainable Alternative for the Cosmetics and Pharmaceutical Industry - A Review.Engineering in life sciences · 2024Review
- Article
- Expanding the genetic toolbox for Cutaneotrichosporon oleaginosus employing newly identified promoters and a novel antibiotic resistance marker.BMC biotechnology · 2023Article
- Expanding the genetic toolbox of Rhodotorula toruloides by identification and validation of six novel promoters induced or repressed under nitrogen starvation.Microbial cell factories · 2023Article
- Article
- Development of a dedicated Golden Gate Assembly Platform (RtGGA) forMetabolic engineering communications · 2022Article
- Impairment of carotenoid biosynthesis through CAR1 gene mutation results in CoQApplied microbiology and biotechnology · 2022Article
- Mining novelFrontiers in microbiology · 2022Article
- Rhodotorula toruloides: an ideal microbial cell factory to produce oleochemicals, carotenoids, and other products.World journal of microbiology & biotechnology · 2021Review
- Understanding and exploiting the fatty acid desaturation system in Rhodotorula toruloides.Biotechnology for biofuels · 2021Article
- Rhodosporidium toruloides - A potential red yeast chassis for lipids and beyond.FEMS yeast research · 2020Review
- A toolset of constitutive promoters for metabolic engineering of Rhodosporidium toruloides.Microbial cell factories · 2019Article
- Multiplexed CRISPR-Cas9-Based Genome Editing ofmSphere · 2019Article
- Screening of endogenous strong promoters for enhanced production of medium-chain-length polyhydroxyalkanoates in Pseudomonas mendocina NK-01.Scientific reports · 2019Article
- Identification of novel genes in the carotenogenic and oleaginous yeast Rhodotorula toruloides through genome-wide insertional mutagenesis.BMC microbiology · 2018Article
- Systems and Synthetic Biology Approaches to Engineer Fungi for Fine Chemical Production.Frontiers in bioengineering and biotechnology · 2018Review
- Engineering global transcription to tune lipophilic properties inBiotechnology for biofuels · 2018Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRed yeast species in the Rhodotorula/Rhodosporidium genus are outstanding producers of triacylglyceride and cell biomass. Metabolic engineering is expected to further enhance the productivity and versatility of these hosts for the production of biobased chemicals and fuels. Promoters with strong activity during oil-accumulation stage are critical tools for metabolic engineering of these oleaginous yeasts.
resultsThe upstream DNA sequences of 6 genes involved in lipid biosynthesis or accumulation in Rhodotorula toruloides were studied by luciferase reporter assay. The promoter of perilipin/lipid droplet protein 1 gene (LDP1) displayed much stronger activity (4-11 folds) than that of glyceraldehyde-3-phosphate dehydrogenase gene (GPD1), one of the strongest promoters known in yeasts. Depending on the stage of cultivation, promoter of acetyl-CoA carboxylase gene (ACC1) and fatty acid synthase β subunit gene (FAS1) exhibited intermediate strength, displaying 50-160 and 20-90% levels of GPD1 promoter, respectively. Interestingly, introns significantly modulated promoter strength at high frequency. The incorporation of intron 1 and 2 of LDP1 (LDP1in promoter) enhanced its promoter activity by 1.6-3.0 folds. Similarly, the strength of ACC1 promoter was enhanced by 1.5-3.2 folds if containing intron 1. The intron 1 sequences of ACL1 and FAS1 also played significant regulatory roles. When driven by the intronic promoters of ACC1 and LDP1 (ACC1in and LDP1in promoter, respectively), the reporter gene expression were up-regulated by nitrogen starvation, independent of de novo oil biosynthesis and accumulation. As a proof of principle, overexpression of the endogenous acyl-CoA-dependent diacylglycerol acyltransferase 1 gene (DGA1) by LDP1in promoter was significantly more efficient than GPD1 promoter in enhancing lipid accumulation.
conclusionIntronic sequences play an important role in regulating gene expression in R. toruloides. Three intronic promoters, LDP1in, ACC1in and FAS1in, are excellent promoters for metabolic engineering in the oleaginous and carotenogenic yeast, R. toruloides.
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