Journal
BOTANY
Volume 87, Issue 6, Pages 643-649Publisher
CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
DOI: 10.1139/B09-033
Keywords
Artemisia annua; 2-alkenal reductase; medium chain dehydrogenase/reductase; gene cloning
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Funding
- National Research Council of Canada's Crop for Enhanced Human Health Program
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The molecular genetics of carbon-carbon double bond reduction in the plant Artemisia annua L. was studied. Expressed sequence tags from this plant were investigated for sequences with similarity to known double-bond reductases. This resulted in the isolation of a cDNA, corresponding to the gene A. annua Dbr1 (Double bond reductase1), encoding a member of the medium chain dehydrogenase/reductase protein superfamily with sequence similarity to tobacco allyl alcohol dehydrogenase. Recombinant A. annua Dbr1 protein was purified from Escherischia coli and shown to catalyze the reduction of the carbon-carbon double bond of 2-alkenals. This activity included the reduction of the double bond at C11-C13 in the artemisinin precursor artemisinic aldehyde, albeit with unnatural stereochemistry. The substrate specificity, product stereochemistry, and expression pattern of A. annua Dbr1 point to its involvement in planta in the detoxification of 2-alkenals, which may be generated under oxidative stress conditions.
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