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Planta

, Volume 217, Issue 1, pp 96–101 | Cite as

Cinnamic acid 4-hydroxylase from cell cultures of the hornwort Anthoceros agrestis

  • Maike Petersen
Original Article

Abstract

Cinnamic acid 4-hydroxylase (EC 1.14.13.11), a cytochrome P450-dependent hydroxylase was for the first time characterized from a hornwort, Anthoceros agrestis Paton (Anthocerotaceae). In suspension cultures of A. agrestis up to 5% of the dry weight was accumulated as rosmarinic acid, a natural product commonly known from higher plants (e.g. species of the Lamiaceae and Boraginaceae). Cinnamic acid 4-hydroxylase is involved in the biosynthesis of rosmarinic acid. The participation of cytochrome P450 was demonstrated by the inhibition of hydroxylase activity by cytochrome c and the inhibition of cinnamic acid hydroxylation in a CO-containing atmosphere, which is partially released by illumination with blue light. The apparent K m values were determined to be at 60 μM and 5 μM for NADPH and cinnamic acid, respectively. A comparatively high hydroxylation activity was seen with NADH as electron donor. While the hydroxylase activity with NADPH was strongly inhibited by the competitive electron acceptor cytochrome c, the activity with NADH was less susceptible, indicating the possibility of different electron-transfer pathways.

Keywords

Anthoceros (Anthocerotaceae) Cell culture Cinnamic acid 4-hydroxylase Cytochrome P450 Hornwort Rosmarinic acid 

Abbreviations

C4H

cinnamic acid 4-hydroxylase

DIECA

sodium diethyldithiocarbaminate

DTT

dithiothreitol

PAL

phenylalanine ammonia-lyase

RA

rosmarinic acid

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Institut für Pharmazeutische BiologiePhilipps-Universität MarburgMarburgGermany

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