Abstract
The sesquiterpenoid artemisinin, isolated these 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 first 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 (Nicotiana tabacum L.) resulted in the expression of an active enzyme and the accumulation of amorpha-4,11-diene ranging from 0.2 to 1.7 ng per g fresh weight.
MeSH terms
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Alkyl and Aryl Transferases / chemistry
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Alkyl and Aryl Transferases / genetics*
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Alkyl and Aryl Transferases / metabolism
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Amino Acid Sequence
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Antimalarials* / isolation & purification
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Artemisia / chemistry
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Artemisia / genetics
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Artemisinins*
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Base Sequence
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Cells, Cultured
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Cloning, Molecular
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DNA, Complementary / genetics
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DNA, Complementary / isolation & purification
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DNA, Complementary / metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Gas Chromatography-Mass Spectrometry
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Gene Expression
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Molecular Sequence Data
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Nicotiana / enzymology*
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Nicotiana / genetics
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Plants, Medicinal
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Plants, Toxic*
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Polymerase Chain Reaction
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RNA / isolation & purification
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Sesquiterpenes* / isolation & purification
Substances
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Antimalarials
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Artemisinins
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DNA, Complementary
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Sesquiterpenes
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RNA
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artemisinin
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Alkyl and Aryl Transferases
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amorpha-4,11-diene synthase