Assay of tryptophan decarboxylase from Catharanthus roseus plant cell cultures by high-performance liquid chromatography.
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.
Center for Bio-Pharmaceutical Sciences, State University of Leiden, The Netherlands.
E J
EJ
Pennings
I
I
Hegger
R
R
van der Heijden
J A
JA
Duine
R
R
Verpoorte
D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000032:analysis*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D010946:Plants, Medicinal / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D014363:Tryptamines
D014363:Tryptamines; D001142:Aromatic-L-Amino-Acid Decarboxylases