Te. Wallaart et al., Amorpha-4,11-diene synthase: cloning and functional expression of a key enzyme in the biosynthetic pathway of the novel antimalarial drug artemisinin, PLANTA, 212(3), 2001, pp. 460-465
The sesquiterpenoid artemisinin, isolated from the plant Artemisia annua L.
, and its semi-synthetic derivatives are a new and very effective group of
antimalarial drugs. A branch point in the biosynthesis of this compound is
the cyclisation of the ubiquitous precursor farnesyl diphosphate into the f
irst specific precursor of artemisinin, namely amorpha-4,11-diene. Here we
describe the isolation of a cDNA clone encoding amorpha-4,11-diene synthase
. The deduced amino acid sequence exhibits the highest identity (50%) with
a putative sesquiterpene cyclase of A. annua. When expressed in Escherichia
coli, the recombinant enzyme catalyses the formation of amorpha-4,11-diene
from farnesyl diphosphate. Introduction of the gene into tobacco (Nicotian
a tabacum L.) resulted in the expression of an active enzyme and the accumu
lation of amorpha-4,11-diene ranging from 0.2 to 1.7 ng per g fresh weight.