4-Hydroxybenzoate 3-geranyltransferase from Lithospermum erythrorhizon: purification of a plant membrane-bound prenyltransferase |
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Authors: | Agnes Mühlenweg Martin Melzer Shu-Ming Li Lutz Heide |
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Institution: | Pharmazeutisches Institut, Pharmazeutische Biologie, Eberhard-Karls-Universit?t Tübingen, Auf der Morgenstelle 8, D-72076 Tübingen, Germany, DE
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Abstract: | Geranyldiphosphate:4-hydroxybenzoate 3-geranyltransferase is a regulatory enzyme in the biosynthesis of shikonin, a phytoalexin
and pharmaceutical produced by cell cultures of Lithospermum erythrorhizon Sieb. et Zucc.. In Linsmaier-Skoog medium, the activity of this enzyme could be enhanced more than 200-fold by addition of
methyl jasmonate, and this culture material was used for the solubilization and purification of the enzyme. Of various detergents
examined, digitonin was the most suitable for the solubilization of the enzyme. The solubilized enzyme was purified 800-fold
by chromatography over diethylaminoethyl (DEAE)-Sephacel, Heparin-Sepharose, Reactive Green 19-Agarose, and Cholic Acid-Agarose.
The purified enzyme required magnesium ions as cofactor and was highly specific for geranyldiphosphate (GPP) and 4-hydroxybenzoate
(4HB) as substrates. The K
m values for 4HB and GPP were calculated by the method of Lineweaver and Burk as 18.4 μM and 13.8 μM, respectively.
Received: 2 July 1997 / Accepted: 14 October 1997 |
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Keywords: | : Geranyltransferase Lithospermum Methyl jasmonate Prenyltransferase Shikonin |
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