Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures

Lei Zhang, Ruxian Ding, Yourong Chai, Mercedes Bonfill, Elisabet Moyano, Kirsi-Marja Oksman-Caldentey, Tiefeng Xu, Yan Pi, Zinan Wang, Hanming Zhang, Guoyin Kai, Zhihua Liao, Xiaofen Sun, Kexuan Tang (Corresponding Author)

    Research output: Contribution to journalArticleScientificpeer-review

    260 Citations (Scopus)


    Scopolamine is a pharmaceutically important tropane alkaloid extensively used as an anticholinergic agent. Here, we report the simultaneous introduction and overexpression of genes encoding the rate-limiting upstream enzyme putrescine N-methyltransferase (PMT) and the downstream enzyme hyoscyamine 6 β-hydroxylase (H6H) of scopolamine biosynthesis in transgenic henbane (Hyoscyamus niger) hairy root cultures. Transgenic hairy root lines expressing both pmt and h6h produced significantly higher (P < 0.05) levels of scopolamine compared with the wild-type and transgenic lines harboring a single gene (pmt or h6h). The best line (T3) produced 411 mg/liter scopolamine, which was over nine times more than that in the wild type (43 mg/liter) and more than twice the amount in the highest scopolamine-producing h6h single-gene transgenic line H11 (184 mg/liter). To our knowledge, this is the highest scopolamine content achieved through genetic engineering of a plant. We conclude that transgenic plants harboring both pmt and h6h possessed an increased flux in the tropane alkaloid biosynthetic pathway that enhanced scopolamine yield, which was more efficient than plants harboring only one of the two genes. It seems that the pulling force of the downstream enzyme (the faucet enzyme) H6H plays a more important role in stimulating scopolamine accumulation in H. niger whereas the functioning of the upstream enzyme PMT is increased proportionally. This study provides an effective approach for large-scale commercial production of scopolamine by using hairy root culture systems as bioreactors.
    Original languageEnglish
    Pages (from-to)6786-6791
    JournalProceedings of the National Academy of Sciences of the United States of America
    Issue number17
    Publication statusPublished - 2004
    MoE publication typeA1 Journal article-refereed


    • Agrobacterium
    • hyoscyamine 6-beta-hydroxylase
    • putrescine
    • N-methyltransferse
    • scopolamine
    • transformation


    Dive into the research topics of 'Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures'. Together they form a unique fingerprint.

    Cite this