Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering |
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Authors: | Katja Jouhikainen Laura Lindgren Tuula Jokelainen Raimo Hiltunen Teemu H Teeri Kirsi-Marja Oksman-Caldentey |
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Institution: | (1) Division of Pharmacognosy, Department of Pharmacy, P.O. Box 56, FIN-00014 University of Helsinki, Finland, FI;(2) Institute of Biotechnology, P.O. Box 56, FIN-00014 University of Helsinki, Finland, FI;(3) VTT Biotechnology and Food Research, Technical Research Centre of Finland, P.O. Box 1500, FIN-02044 VTT, Finland, FI |
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Abstract: | In order to test the possibility of enhancing the production of pharmaceutically valuable scopolamine in transgenic cultures,
the 35S-h6h transgene that codes for the enzyme hyoscyamine-6β-hydroxylase (EC 1.14.11.11) was introduced into Hyoscyamus muticus L. strain Cairo (Egyptian henbane). This plant was chosen for its capability to produce very high amounts of tropane alkaloids
(up to 6% of the dry weight in the leaves of mature plant). To our knowledge, this is the first time such a large population
of transgenic cultures has been studied at the morphological, chemical and genetic levels. A great variation was observed
in the tropane alkaloid production among the 43 positive transformants. The best clone (KB7) produced 17 mg/l scopolamine,
which is over 100 times more than the control clones. However, conversion of hyoscyamine to scopolamine was still incomplete.
The expression of h6h was found to be proportional to the scopolamine production, and was the main reason behind the variation in the scopolamine/hyoscyamine
ratio in the hairy-root clones. These results indicate that H. muticus strain Cairo has a potential for even more enhanced scopolamine production with more efficient gene-expression systems.
Received: 24 December 1998 / Accepted: 13 January 1999 |
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Keywords: | : Genetic engineering Hairy root Hyoscyamine-6β -hydroxylase Hyoscyamus Scopolamine Tropane alkaloid |
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