TY - JOUR
T1 - Amorpha-4,11-diene synthase catalyses the first probable step in artemisinin biosynthesis
AU - Bouwmeester, Harro J.
AU - Wallaart, T. Eelco
AU - Janssen, Michiel H.A.
AU - Van Loo, Bert
AU - Jansen, Ben J.M.
AU - Posthumus, Maarten A.
AU - Schmidt, Claus O.
AU - De Kraker, Jan Willem
AU - König, Wilfried A.
AU - Franssen, Maurice C.R.
PY - 1999/11/1
Y1 - 1999/11/1
N2 - The endoperoxide sesquiterpene lactone artemisinin and its derivatives are a promising new group of drugs against malaria. Artemisinin is a constituent of the annual herb Artemisia annua L. So far only the later steps in artemisinin biosynthesis - from artemisinic acid - have been elucidated and the expected olefinic sesquiterpene intermediate has never been demonstrated. In pentane extracts of A. annua leaves we detected a sesquiterpene with the mass spectrum of amorpha-4,11-diene. Synthesis of amorpha-4,11-diene from artemisinic acid confirmed the identity. In addition we identified several sesquiterpene synthases of which one of the major activities catalysed the formation of amorpha-4,11-diene from farnesyl diphosphate. This enzyme was partially purified and shows the typical characteristics of sesquiterpene synthases, such as a broad pH optimum around 6.5-7.0, a molecular mass of 56 kDa, and a K(m) of 0.6 μM. The structure and configuration of amorpha-4,11-diene, its low content in A. annua and the high activity of amorpha-4,11-diene synthase all support that amorpha-4,11-diene is the likely olefinic sesquiterpene intermediate in the biosynthesis of artemisinin.
AB - The endoperoxide sesquiterpene lactone artemisinin and its derivatives are a promising new group of drugs against malaria. Artemisinin is a constituent of the annual herb Artemisia annua L. So far only the later steps in artemisinin biosynthesis - from artemisinic acid - have been elucidated and the expected olefinic sesquiterpene intermediate has never been demonstrated. In pentane extracts of A. annua leaves we detected a sesquiterpene with the mass spectrum of amorpha-4,11-diene. Synthesis of amorpha-4,11-diene from artemisinic acid confirmed the identity. In addition we identified several sesquiterpene synthases of which one of the major activities catalysed the formation of amorpha-4,11-diene from farnesyl diphosphate. This enzyme was partially purified and shows the typical characteristics of sesquiterpene synthases, such as a broad pH optimum around 6.5-7.0, a molecular mass of 56 kDa, and a K(m) of 0.6 μM. The structure and configuration of amorpha-4,11-diene, its low content in A. annua and the high activity of amorpha-4,11-diene synthase all support that amorpha-4,11-diene is the likely olefinic sesquiterpene intermediate in the biosynthesis of artemisinin.
KW - Amorpha-4,11-diene
KW - Artemisia annua
KW - Artemisinin
KW - Asteraceae
KW - Biosynthesis
KW - Sesquiterpenoids
UR - http://www.scopus.com/inward/record.url?scp=0033230281&partnerID=8YFLogxK
U2 - 10.1016/S0031-9422(99)00206-X
DO - 10.1016/S0031-9422(99)00206-X
M3 - Article
C2 - 10626375
AN - SCOPUS:0033230281
SN - 0031-9422
VL - 52
SP - 843
EP - 854
JO - Phytochemistry
JF - Phytochemistry
IS - 5
ER -
