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Co-expression of squalene epoxidases with triterpene cyclases boosts production of triterpenoids in plants and yeast

(2018) METABOLIC ENGINEERING. 49. p.1-12
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Abstract
Triterpene cyclases catalyze the first committed step in triterpene biosynthesis, by forming mono- to pentacyclic backbone structures from oxygenated C30 isoprenoid precursors. Squalene epoxidase precedes this cyclization by providing the oxygenated and activated substrate for triterpene biosynthesis. Three squalene epoxidases from Cucurbita pepo (CpSEs) were isolated and shown to have evolved under purifying selection with signs of sites under positive selection in their N- and C-termini. They all localize to the Endoplasmic Reticulum (ER) and produce 2,3-oxidosqualene and 2,3: 22,23-dioxidosqualene when expressed in a yeast erg1 (squalene epoxidase) erg7 (lanosterol synthase) double mutant. Co-expression of the CpSEs with four different triterpene cyclases, either transiently in Nicotiana benthamiana or constitutively in yeast, showed that CpSEs boost triterpene production. CpSE2 was the best performing in this regard, which could reflect either increased substrate production or superior channeling of the substrate to the triterpene cyclases. Fluorescence Lifetime Imaging Microscopy (FLIM) analysis with C. pepo cucurbitadienol synthase (CpCPQ) revealed a specific interaction with CpSE2 but not with the other CpSEs. When CpSE2 was transformed into C. pepo hairy root lines, cucurbitacin E production was increased two folds compared to empty vector control lines. This study provides new insight into the importance of SEs in triterpene biosynthesis, suggesting that they may facilitate substrate channeling, and demonstrates that SE overexpression is a new tool for increasing triterpene production in plants and yeast.
Keywords
Cucurbitacin, Subcellular localization, Squalene epoxidase, Metabolic engineering, Triterpene, Protein-protein interaction, COENZYME-A REDUCTASE, BETA-AMYRIN SYNTHASE, SACCHAROMYCES-CEREVISIAE, SAPONIN BIOSYNTHESIS, MEDICAGO-TRUNCATULA, HIGH-THROUGHPUT, PHYTOSTEROL BIOSYNTHESIS, OXIDOSQUALENE CYCLASE, DIPHOSPHATE SYNTHASE, ENHANCED TRITERPENE

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Chicago
Dong, Lemeng, Jacob Pollier, Jean-Etienne Bassard, Georgios Ntallas, Aldo Almeida, Eleni Lazaridi, Bekzod Khakimov, et al. 2018. “Co-expression of Squalene Epoxidases with Triterpene Cyclases Boosts Production of Triterpenoids in Plants and Yeast.” Metabolic Engineering 49: 1–12.
APA
Dong, L., Pollier, J., Bassard, J.-E., Ntallas, G., Almeida, A., Lazaridi, E., Khakimov, B., et al. (2018). Co-expression of squalene epoxidases with triterpene cyclases boosts production of triterpenoids in plants and yeast. METABOLIC ENGINEERING, 49, 1–12.
Vancouver
1.
Dong L, Pollier J, Bassard J-E, Ntallas G, Almeida A, Lazaridi E, et al. Co-expression of squalene epoxidases with triterpene cyclases boosts production of triterpenoids in plants and yeast. METABOLIC ENGINEERING. 2018;49:1–12.
MLA
Dong, Lemeng et al. “Co-expression of Squalene Epoxidases with Triterpene Cyclases Boosts Production of Triterpenoids in Plants and Yeast.” METABOLIC ENGINEERING 49 (2018): 1–12. Print.
@article{8586123,
  abstract     = {Triterpene cyclases catalyze the first committed step in triterpene biosynthesis, by forming mono- to pentacyclic backbone structures from oxygenated C30 isoprenoid precursors. Squalene epoxidase precedes this cyclization by providing the oxygenated and activated substrate for triterpene biosynthesis. Three squalene epoxidases from Cucurbita pepo (CpSEs) were isolated and shown to have evolved under purifying selection with signs of sites under positive selection in their N- and C-termini. They all localize to the Endoplasmic Reticulum (ER) and produce 2,3-oxidosqualene and 2,3: 22,23-dioxidosqualene when expressed in a yeast erg1 (squalene epoxidase) erg7 (lanosterol synthase) double mutant. Co-expression of the CpSEs with four different triterpene cyclases, either transiently in Nicotiana benthamiana or constitutively in yeast, showed that CpSEs boost triterpene production. CpSE2 was the best performing in this regard, which could reflect either increased substrate production or superior channeling of the substrate to the triterpene cyclases. Fluorescence Lifetime Imaging Microscopy (FLIM) analysis with C. pepo cucurbitadienol synthase (CpCPQ) revealed a specific interaction with CpSE2 but not with the other CpSEs. When CpSE2 was transformed into C. pepo hairy root lines, cucurbitacin E production was increased two folds compared to empty vector control lines. This study provides new insight into the importance of SEs in triterpene biosynthesis, suggesting that they may facilitate substrate channeling, and demonstrates that SE overexpression is a new tool for increasing triterpene production in plants and yeast.},
  author       = {Dong, Lemeng and Pollier, Jacob and Bassard, Jean-Etienne and Ntallas, Georgios and Almeida, Aldo and Lazaridi, Eleni and Khakimov, Bekzod and Arendt, Philipp and Souza de Oliveira, Louisi and Lota, Frederic and Goossens, Alain and Michoux, Franck and Bak, Soren},
  issn         = {1096-7176},
  journal      = {METABOLIC ENGINEERING},
  language     = {eng},
  pages        = {1--12},
  title        = {Co-expression of squalene epoxidases with triterpene cyclases boosts production of triterpenoids in plants and yeast},
  url          = {http://dx.doi.org/10.1016/j.ymben.2018.07.002},
  volume       = {49},
  year         = {2018},
}

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