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An assay for secologanin in plant tissues based on enzymatic conversion into strictosidine
Author(s) -
Hallard Didier,
van der Heijden Robert,
Contin Adriana,
Jiménéz Emilia M. Tomas,
Snoeijer Wim,
Verpoorte Robert,
Jensen Søren R.,
Cardoso M. Inês Lopes,
Pasquali Giancarlo,
Memelink Johan,
Hoge J. Harry C.
Publication year - 1998
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/(sici)1099-1565(199807/08)9:4<162::aid-pca394>3.0.co;2-d
Subject(s) - tryptamine , catharanthus roseus , chemistry , indole test , biochemistry , chromatography
The secoiridoid glucoside secologanin is the terpenoid building block in the biosynthesis of terpenoid indole alkaloids. A method for its determination in plant tissues and in cell suspension cultures has been developed. This assay is based on the condensation of secologanin with tryptamine, yielding strictosidine, in a reaction catalysed by the enzyme strictosidine synthase (STR; E.C. 4.3.3.2). Subsequently, the formation of strictosidine is quantified by high performance liquid chromatography (HPLC). STR was isolated from transgenic Nicotiana tabacum cells expressing a cDNA‐derived gene coding for STR from Catharanthus roseus . The high specificity of STR for secologanin, in combination with a sensitive and selective HPLC system, allows a simple extraction of secologanin from plant tissue. The detection limit of this method is 15 ng secologanin. Using this assay, secologanin contents were determined in tissues of various plant species; Lonicera xylosteum hairy roots were found to contain 1% of secologanin on a dry weight basis. © 1998 John Wiley & Sons, Ltd.