Engineering Escherichia coli for the synthesis of taxadiene, a key intermediate in the biosynthesis of taxol

Taxadiene, the key intermediate of paclitaxel (Taxol) biosynthesis, has bee n prepared enzymatically from isopentenyl diphosphate in cell-free extracts of Escherichia coli by overexpressing genes encoding isopentenyl diphospha te isomerase, geranylgeranyl diphosphate synthase and taxadiene synthase. I n addition, by the expression of three genes encoding four enzymes on the t erpene biosynthetic pathway in a single strain of E. coli, taxadiene can be conveniently synthesized in vivo, at the unoptimized yield of 1.3 mg per l iter of cell culture. The success of both in vitro and in vivo synthesis of taxadiene bodes well for the future production of taxoids by non-paclitaxe l producing organisms through pathway engineering. (C) 2001 Elsevier Scienc e Ltd. All rights reserved.