Planta Med 2002; 68(10): 906-911
DOI: 10.1055/s-2002-34935
Original Paper
Biochemistry, Physiologie, in vitro-cultures
© Georg Thieme Verlag Stuttgart · New York

A Newly-Detected Reductase from Rauvolfia Closes a Gap in the Biosynthesis of the Antiarrhythmic Alkaloid Ajmaline

Shujuan Gao1 [1] , Gerald von Schumann1 [1] , Joachim Stöckigt1
  • 1Johannes Gutenberg-University Mainz, Institute of Pharmacy, Department of Pharmaceutical Biology, Germany
Further Information

Publication History

Received: January 10, 2002

Accepted: May 11, 2002

Publication Date:
21 October 2002 (online)

Abstract

A new enzyme, 1,2-dihydrovomilenine reductase (E.C. 1.3.1), has been detected in Rauvolfia cell suspension cultures. The enzyme specifically converts 2β(R)-1,2-dihydrovomilenine through an NADPH-dependent reaction into 17-O-acetylnorajmaline, a close biosynthetic precursor of the antiarrhythmic alkaloid ajmaline from Rauvolfia. A five-step purification procedure using SOURCE 30Q chromatography, hydroxyapatite chromatography, 2′,5′-ADP Sepharose 4B affinity chromatography and ion exchange chromatography on DEAE Sepharose and Mono Q delivered a ∼ 200-fold enriched enzyme in a yield of ∼ 6 %. SDS-PAGE showed an Mr for the enzyme of ∼ 48 kDa. Optimum pH and optimum temperature of the reductase were at pH 6.0 and 37 °C. The enzyme shows a limited distribution in cell cultures expressing ajmaline biosynthesis, and is obviously highly specific for the ajmaline pathway.

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1 Shujuan Gao and Gerald von Schumann have contributed equally to this study.

Prof. Dr. J. Stöckigt

Department of Pharmaceutical Biology

Institute of Pharmacy

Johannes Gutenberg-University

Staudinger Weg 5

55099 Mainz, Germany

Email: stoeckig@mail.uni-mainz.de

Phone: +49-(0)6131-39-25751

Fax: +49-(0)6131-39-23752