Biosynthesis of Taxol: Enzymatic Acetylation of 10-Deacetylbaccatin-III to Baccatin-III in Crude Extracts from Roots of Taxus baccata

Authors: Zocher R.¹; Weckwerth W.¹; Hacker C.¹; Kammer B.¹; Hornbogen T.¹; Ewald D.²

Source: Biochemical and Biophysical Research Communications, Volume 229, Number 1, December 1996 , pp. 16-20(5)

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Abstract:

The biosynthesis of taxol is a multistep process. One intermediate reaction is the acetylation of 10-deacetylbaccatin-III (10-DAB) to baccatin-III, an assumed precursor of taxol. Here we describe the cell free acetylation of 10-DAB in crude extracts from roots of Taxus baccata saplings using 14 C- or 3 H-labeled acetyl-coenzyme A as the acetyl donor. The reaction is strictly dependent on the addition of 10-DAB and is specific for the 10-hydroxyl group of the taxane ring. Formation of radiolabeled baccatin-III was confirmed by co-chromatography of the labeled product with authentic baccatin-III in different TLC-systems and HPLC. Furthermore, the acetylation product showed an identical UV spectrum as authentic baccatin-III. Crude extracts from cambium of stems yielded three- to fivefold lower activity. This is in agreement with our finding that the taxol titer in roots was considerably higher than that in cambium.

Language: English

Document Type: Research article

Affiliations: 1: Institut fur Biochemie und Molekulare Biologie, Technische Universitat Berlin, Franklinstrasse 29, Berlin, D-10587, Germany 2: Institut fur Forstpflanzenzuchtung, Bundesforschungsanstalt fur Forst- und Holzwirtschaft, Eberswalder Chaussee 6, Waldsieversdorf, D-15377, Germany

Publication date: 1996-12-01