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Modification of isoprene synthesis to enable production of curcurbitadienol synthesis in Saccharomyces cerevisiae

  • Jing Qiao
  • Zuliang Luo
  • Shengrong Cui
  • Huan Zhao
  • Qi Tang
  • Changming Mo
  • Xiaojun MaEmail author
  • Zimian DingEmail author
Fermentation, Cell Culture and Bioengineering - Original Paper
  • 172 Downloads

Abstract

Cucurbitane-type triterpenoids such as mogrosides and cucurbitacins that are present in the plants of Cucurbitaceae are widely used in Asian traditional medicine. Cucurbitadienol is the skeleton of cucurbitane-type triterpenoids. As an alternative production strategy, we developed baker’s yeast Saccharomyces cerevisiae as a microbial host for the eventual transformation of cucurbitadienol. The synthetic pathway of cucurbitadienol was constructed in Saccharomyces cerevisiae by introducing the cucurbitadienol synthase gene from different plants, resulting in 7.80 mg cucurbitadienol from 1 L of fermentation broth. Improving supplies of isoprenoid precursors was then investigated for increasing cucurbitadienol production. Cucurbitadienol production increased to 21.47 mg/L through the overexpression of a global regulatory factor (UPC2) gene of triterpenoid synthase. In addition, knockout of the ERG7 gene increased cucurbitadienol production from 21.47 to 61.80 mg/L. Finally, fed-batch fermentation was performed, and 63.00 mg/L cucurbitadienol was produced. This work is an important step towards the total biosynthesis of valuable cucurbitane-type triterpenoids and demonstrates the potential for developing a sustainable and secure yeast biomanufacturing platform for triterpenoids.

Keywords

Triterpenoids Cucurbitadienol Metabolic engineering Saccharomyces cerevisiae 

Notes

Acknowledgements

We thank Dr. Yuan Zhou (Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences) for providing the cucurbitadienol standard.

Funding

This work was supported by the National Natural Science Foundation of China (NO. 81573521), Beijing Municipal Natural Science Foundation (NO. 5172028) and CAMS Innovation Fund for Medical Sciences (NO. 2017-I2M-1-013).

Compliance with ethical standards

Conflict of interest

Authors declare that they have no conflicts of interest.

Supplementary material

10295_2018_2116_MOESM1_ESM.docx (134 kb)
Supplementary material 1 (DOCX 134 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2018

Authors and Affiliations

  • Jing Qiao
    • 1
  • Zuliang Luo
    • 1
  • Shengrong Cui
    • 1
  • Huan Zhao
    • 2
  • Qi Tang
    • 3
  • Changming Mo
    • 4
  • Xiaojun Ma
    • 1
    Email author
  • Zimian Ding
    • 1
    Email author
  1. 1.Institute of Medicinal Plant DevelopmentChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina
  2. 2.State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia MedicaAcademy of Chinese Medical SciencesBeijingChina
  3. 3.National and Local Union Engineering Research Center of Veterinary Herbal Medicine Resources and Initiative and Hunan Co-Innovation Center for Utilization of Botanical Functional IngredientsHunan Agricultural UniversityChangshaChina
  4. 4.Guangxi Botanical Garden of Medicinal PlantsNanningChina

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