ArticleApplied and environmental microbiology2000
Autocloning and amplification of LIP2 in Yarrowia lipolytica.
Article in Applied and environmental microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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39 citing papers in PubMed, 147 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
- YeastInternational journal of molecular sciences · 2026Article
- New Yarrowia lipolytica chassis strains for industrial enzyme production.Microbial cell factories · 2025Article
- Improving and Streamlining Gene Editing inJournal of fungi (Basel, Switzerland) · 2024Article
- Engineering 3-Hydroxypropionic Acid Production from Glucose inJournal of fungi (Basel, Switzerland) · 2023Article
- Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system.FEMS yeast research · 2023Article
- A Novel Cre/International journal of molecular sciences · 2021Article
- Review
- Impact of overproduced heterologous protein characteristics on physiological response in Yarrowia lipolytica steady-state-maintained continuous cultures.Applied microbiology and biotechnology · 2020Article
- Optimization of Yarrowia lipolytica-based consolidated biocatalyst through synthetic biology approach: transcription units and signal peptides shuffling.Applied microbiology and biotechnology · 2020Article
- A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity.Biotechnology letters · 2020Article
- An artificial chromosome ylAC enables efficient assembly of multiple genes in Yarrowia lipolytica for biomanufacturing.Communications biology · 2020Article
- Comprehensive comparison of Yarrowia lipolytica and Pichia pastoris for production of Candida antarctica lipase B.Scientific reports · 2020Article
- A modular Golden Gate toolkit for Yarrowia lipolytica synthetic biology.Microbial biotechnology · 2019Article
- Efficient expression vectors and host strain for the production of recombinant proteins by Yarrowia lipolytica in process conditions.Microbial cell factories · 2019Article
- Noncovalent Immobilization ofBiomolecules · 2019Article
- Increase of Candida antarctica lipase B production under PGK promoter in Pichia pastoris: effect of multicopies.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2019Article
- The metabolism and genetic regulation of lipids in the oleaginous yeast Yarrowia lipolytica.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2019Review
- Filamentous fungi-like secretory pathway strayed in a yeast system: peculiarities of Yarrowia lipolytica secretory pathway underlying its extraordinary performance.Applied microbiology and biotechnology · 2019Review
- Synthetic biology tools for engineering Yarrowia lipolytica.Biotechnology advances · 2018Review
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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
We synthesized a Yarrowia lipolytica strain overproducing lipase for industrial applications by using long terminal repeat (zeta) of the Y. lipolytica retrotransposon Ylt1 and an allele of URA3 with a promoter deletion to construct JMP3. JMP3 is a derivative of plasmid pHSS6 carrying a NotI-NotI cassette which contains a defective URA3 allele, a polylinker sequence, and the zeta region for targeting to multiple sites in the genome of the recipient. We inserted the LIP2 gene (encoding extracellular lipase) under the control of the strong POX2 promoter into JMP3 to generate JMP6. The pHSS6 region was removed by NotI digestion prior to transformation. Two Y. lipolytica strains transformed with the JMP6 LIP2 cassette had a mean of 10 integrated copies devoid of the Escherichia coli region, corresponding to an autocloning event. The copy number in the transformants was stable even after 120 generations in nonselective and lipase-inducing conditions. The resulting strains could produce 0.5 g of active lipase per liter in the supernatant, 40 times more than the single-copy strain with the LIP2 promoter. This work provides a new expression system in Y. lipolytica that results in strains devoid of bacterial DNA and in strains producing a high level of lipase for industrial uses, waste treatment, and pancreatic insufficiency therapy.
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