The molecular cloning of artemisinic aldehyde Delta11(13) reductase and its role in glandular trichome-dependent biosynthesis of artemisinin in Artemisia annua.
At some point during biosynthesis of the antimalarial artemisinin in glandular trichomes of Artemisia annua, the Delta11(13) double bond originating in amorpha-4,11-diene is reduced. This is thought to occur in artemisinic aldehyde, but other intermediates have been suggested. In an effort to understand double bond reduction in artemisinin biosynthesis, extracts of A. annua flower buds were investigated and found to contain artemisinic aldehyde Delta11(13) double bond reductase activity. Through a combination of partial protein purification, mass spectrometry, and expressed sequence tag analysis, a cDNA clone corresponding to the enzyme was isolated. The corresponding gene Dbr2, encoding a member of the enoate reductase family with similarity to plant 12-oxophytodienoate reductases, was found to be highly expressed in glandular trichomes. Recombinant Dbr2 was subsequently characterized and shown to be relatively specific for artemisinic aldehyde and to have some activity on small alpha,beta-unsaturated carbonyl compounds. Expression in yeast of Dbr2 and genes encoding four other enzymes in the artemisinin pathway resulted in the accumulation of dihydroartemsinic acid. The relevance of Dbr2 to trichome-specific artemisinin biosynthesis is discussed.
Zhang
Yansheng
Y
Plant Biotechnology Institute, 110 Gymnasium Pl., Saskatoon, Saskatchewan, Canada.
Teoh
Keat H
KH
Reed
Darwin W
DW
Maes
Lies
L
Goossens
Alain
A
Olson
Douglas J H
DJ
Ross
Andrew R S
AR
Covello
Patrick S
PS
D031172:Artemisia annua / Q000201:enzymology*; D037621:Artemisinins / Q000737:chemistry*; D002244:Carbon / Q000737:chemistry; D003001:Cloning, Molecular; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D005822:Genetic Vectors; D008956:Models, Chemical; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics*; D018517:Plant Roots / Q000378:metabolism; D012639:Seeds; D013237:Stereoisomerism; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors
D037621:Artemisinins; D010940:Plant Proteins; D002244:Carbon; C031327:artemisinin; D010088:Oxidoreductases; C531287:artemisinic aldehyde delta11(13) reductase, Artemisia annua