Biosynthesis of pseudoisoeugenols in tissue cultures of Pimpinella anisum. Phenylalanine ammonia lyase and cinnamic acid 4-hydroxylase activities.
The genus Pimpinella contains pseudoisoeugenols, phenylpropanoids with a rare 2,5-dioxy substitution pattern on the phenyl ring. To study the biosynthesis of these compounds, we set up a leaf-differentiating tissue culture of Pimpinella anisum. These cultures mainly produce epoxy-pseudoisoeugenol-(2-methylbutyrate). To corroborate the biosynthetic pathway of epoxy-pseudoisoeugenol-(2-methylbutyrate) as proposed on the basis of investigations with 13C/14C-labelled precursors, the key steps of the pathway were investigated at an enzyme level. Experiments with cell-free homogenates clearly revealed that L-phenylalanine is converted to (E)-cinnamic acid by phenylalanine ammonia lyase and that (E)-cinnamic acid is converted to p-coumaric acid by cinnamic acid 4-hydroxylase. L-2-aminooxy-3-phenylpropionic acid, an analogue of L-phenylalanine, inhibited the incorporation of L-[3'-13C]phenylalanine into epoxy-pseudoisoeugenol-(2-methylbutyrate). Up to 2% of the precursor DL-[3'-13C]phenyllactate was incorporated into epoxy-pseudoisoeugenol-(2-methylbutyrate). Inhibition experiments with oxalacetic acid clearly showed that cinnamic acid is not formed by dehydration of phenyllactic acid in this leaf-differentiating tissue culture of P. anisum.
Reichling
J
J
Institut für Pharmazeutische Biologie, Universität Heidelberg, Germany.
Kemmerer
B
B
Sauer-Gürth
H
H
D002474:Cell-Free System; D002934:Cinnamates / Q000378:metabolism*; D004789:Enzyme Activation; D019684:Magnoliopsida / Q000166:cytology / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism*; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis*; D059828:Plant Cells; D010944:Plants / Q000378:metabolism*; D013997:Time Factors
D002934:Cinnamates; C029010:cinnamic acid; D010649:Phenylalanine; D010650:Phenylalanine Ammonia-Lyase