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The isolation and characterization of dammarenediol synthase gene from Panax quinquefolius and its heterologous co-expression with cytochrome P450 gene PqD12H in yeast

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Abstract

Panax quinquefolius is one of perennial herbs and well known for its outstanding pharmacological activity. Ginsenosides are thought to be the main active ingredients in Panax quinquefolius and exist in many kinds of plant genus Panax (ginseng). Dammarenediol synthase, which is considered as a key enzyme in ginsenoside biosynthesis pathway can convert 2, 3-oxidosqualene into dammarenediol-II. However, the dammarenediol synthase gene in Panax quinquefolius has not been identified. Here, we cloned and identified a dammarenediol synthase gene from Panax quinquefolius (PqDS, GenBank accession No. KC316048) at the first time, and reverse transcription-PCR (RT-PCR) analysis also showed an obvious transcription increase of PqDS in the methyl jasmonate (MeJA)-induced hairy roots. Ectopic expression of PqDS in yeast resulted in the production of dammarenediol-II was confirmed by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Moreover, overexpression of PqDS in transgenic hairy roots could increase the transcription of gene PqDS and another P450 gene PqD12H (encoding protopanaxadiol synthase in Panax quinquefolius), the accumulation of ginsenosides also increased at the same time. In addition, both PqDS and PqD12H gene co-expressed in recombinant yeast result in the production of protopanaxadiol was detected by LC/APCIMS; this result also provides a new strategy for the abundant production of protopanaxadiol in vitro.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863), NO. 2013AA102604, Projects of National Science Foundation of China, NO. 30970259. 31270337, Research Fund for the Doctoral Program of Higher Education of China, NO. 20120061110038, and Scientific and Technological Development Plan Project of Jilin Province, NO. 20130102041JC.

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Authors and Affiliations

  1. School of Biological and Agricultural Engineering, Jilin University, No.5988 Renmin Street Nanguan District, Changchun, Peoples’ Republic of China

    Le Wang, Shou-Jing Zhao & Hao-Jie Cao

  2. The School of Biological and Agricultural Engineering, Changchun University of Technology, No. 2055 Yanan Street, Changchun, Peoples’ Republic of China

    Yao Sun

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  1. Le Wang
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  2. Shou-Jing Zhao
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  3. Hao-Jie Cao
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Correspondence to Shou-Jing Zhao.

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Hao-Jie Cao and Yao Sun assisted to this work.

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Wang, L., Zhao, SJ., Cao, HJ. et al. The isolation and characterization of dammarenediol synthase gene from Panax quinquefolius and its heterologous co-expression with cytochrome P450 gene PqD12H in yeast. Funct Integr Genomics 14, 545–557 (2014). https://doi.org/10.1007/s10142-014-0384-1

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  • DOI: https://doi.org/10.1007/s10142-014-0384-1

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