Assay of tryptophan decarboxylase from Catharanthus roseus plant cell cultures by high-performance liquid chromatography |
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| Authors: | E J Pennings I Hegger R van der Heijden J A Duine R Verpoorte |
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| Affiliation: | Center for Bio-Pharmaceutical Sciences, State University of Leiden, The Netherlands. |
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| Abstract: | An assay is described for the enzyme tryptophan decarboxylase from plant cell suspension cultures. It is based on the fluorometric detection of tryptamine by HPLC on a LiChrosorb RP-8 Select B column. Tryptophan decarboxylase from Catharanthus roseus was induced by transferring 14-day-old cells into an induction medium. Optimum activity was found 2 days after transfer, the increase being 5- to 10-fold. When kept at -15 degrees C the crude enzyme lost half its activity in about 7 days. The rate of the decarboxylation reaction was linear for at least 3 h at 35 degrees C. |
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