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Production of the dammarene sapogenin (protopanaxadiol) in transgenic tobacco plants and cultured cells by heterologous expression of PgDDS and CYP716A47

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Abstract

Key message

Protopanaxadiol (PPD) is an aglycone of dammarene-type ginsenoside and has high medicinal values. In this work, we reported the PPD production in transgenic tobacco co-overexpressing PgDDS and CYP716A47.

Abstract

PPD is an aglycone of ginsenosides produced by Panax species and has a wide range of pharmacological activities. PPD is synthesized via the hydroxylation of dammarenediol-II (DD) by CYP716A47 enzyme. Here, we established a PPD production system via cell suspension culture of transgenic tobacco co-overexpressing the genes for PgDDS and CYP716A47. The concentration of PPD in transgenic tobacco leaves was 2.3–5.7 µg/g dry weight (DW), depending on the transgenic line. Leaf segments were cultured on medium with various types of hormones to induce callus. Auxin treatment, particularly 2,4-D, strongly enhanced the production of DD (783.8 µg g−1 DW) and PPD (125.9 µg g−1 DW). Treatment with 2,4-D enhanced the transcription of the HMG-Co reductase (HMGR) and squalene epoxidase genes. PPD production reached 166.9 and 980.9 µg g−1 DW in a 250-ml shake flask culture and in 5-l airlift bioreactor culture, respectively.

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Abbreviations

BAR:

Basta

DD:

Dammarenediol-II

PPD:

Protopanaxadiol

DDS:

Dammarenediol-II synthase

PgDDS:

Panax ginseng dammarenediol-II synthase

CYP716A47:

Cytochrome P450 716A47

qPCR:

Real-time polymerase chain reaction

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Acknowledgments

This work was supported by the Rural Development Administration, Republic of Korea [Next-Generation BioGreen 21 Program (PJ011285)], and by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2013R1A1A2064352).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chunchon, 200-701, Republic of Korea

    Ju-Hyeon Chun, Prakash Babu Adhikari, Seong-Bum Park, Jung-Yeon Han & Yong-Eui Choi

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  1. Ju-Hyeon Chun
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  2. Prakash Babu Adhikari
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  3. Seong-Bum Park
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  4. Jung-Yeon Han
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  5. Yong-Eui Choi
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Correspondence to Yong-Eui Choi.

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Communicated by F. Sato.

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Chun, JH., Adhikari, P.B., Park, SB. et al. Production of the dammarene sapogenin (protopanaxadiol) in transgenic tobacco plants and cultured cells by heterologous expression of PgDDS and CYP716A47 . Plant Cell Rep 34, 1551–1560 (2015). https://doi.org/10.1007/s00299-015-1806-9

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  • DOI: https://doi.org/10.1007/s00299-015-1806-9

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