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Metabolic engineering of Saccharomyces cerevisiae for production of germacrene A, a precursor of beta-elemene

  • Metabolic Engineering and Synthetic Biology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Beta-elemene, a sesquiterpene and the major component of the medicinal herb Curcuma wenyujin, has antitumor activity against various types of cancer and could potentially serve as a potent antineoplastic drug. However, its current mode of production through extraction from plants has been inefficient and suffers from limited natural resources. Here, we engineered a yeast cell factory for the sustainable production of germacrene A, which can be transformed to beta-elemene by a one-step chemical reaction in vitro. Two heterologous germacrene A synthases (GASs) converting farnesyl pyrophosphate (FPP) to germacrene A were evaluated in yeast for their ability to produce germacrene A. Thereafter, several metabolic engineering strategies were used to improve the production level. Overexpression of truncated 3-hydroxyl-3-methylglutaryl-CoA reductase and fusion of FPP synthase with GAS, led to a sixfold increase in germacrene A production in shake-flask culture. Finally, 190.7 mg/l of germacrene A was achieved. The results reported in this study represent the highest titer of germacrene A reported to date. These results provide a basis for creating an efficient route for further industrial application re-placing the traditional extraction of beta-elemene from plant sources.

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Acknowledgements

This work was supported by the National Science Fund for Distinguished Young Scholars (81325023). Part of this work was funded by the Knut and Alice Wallenberg foundation and the Novo Nordisk Foundation. We thank Yun Chen, Min Chen and Ping Su for providing good comments and help. We also thank Guodong Liu for reading the manuscript and providing valuable suggestions.

Authors’ contribution

JN and LQH conceived and supervised the study. YTH designed and performed the experiments as well as drafting the manuscript. AK assisted with the construction of plasmid pBS01 and participated in experiment design. YJZ participated in the design and provided many valuable suggestions during the study. JCB contributed to Gibson cloning and fermentation experiment. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijng, 100700, People’s Republic of China

    Yating Hu & Luqi Huang

  2. Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-412 96, Gothenburg, Sweden

    Yating Hu, Yongjin J. Zhou, Jichen Bao, Jens Nielsen & Anastasia Krivoruchko

  3. Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Kemivägen 10, SE-412 96, Gothenburg, Sweden

    Jens Nielsen & Anastasia Krivoruchko

  4. Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2970, Hørsholm, Denmark

    Jens Nielsen

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  1. Yating Hu
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  2. Yongjin J. Zhou
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  3. Jichen Bao
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  4. Luqi Huang
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  6. Anastasia Krivoruchko
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Correspondence to Luqi Huang or Jens Nielsen.

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Hu, Y., Zhou, Y.J., Bao, J. et al. Metabolic engineering of Saccharomyces cerevisiae for production of germacrene A, a precursor of beta-elemene. J Ind Microbiol Biotechnol 44, 1065–1072 (2017). https://doi.org/10.1007/s10295-017-1934-z

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  • DOI: https://doi.org/10.1007/s10295-017-1934-z

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