Amorpha-4,11-diene synthase catalyses the first probable step in artemisinin biosynthesis

Bouwmeester, Harro J.; Wallaart, Thorvald; Janssen, Michiel H. A.; Loo, Bert van; Jansen, B. J. M.; Posthumus, M. A.; Schmidt, Christiane Sybille; Kraker, J.W. de; König, Wilfried Α.; Franssen, M.C.R.

doi:10.1016/s0031-9422(99)00206-x

Cited by 245 publications

(207 citation statements)

References 35 publications

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“…The identity of compound 14 was unambiguously proven by semi-synthesis from artemisinic acid. On the HP-INNOwax column compound 14 had the same retention time and mass spectrum as the synthesized product ( Figure 3) (Bouwmeester et al 1999b). …”

Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 81%

“…Compound 14 is a minor compound that co-eluted with the major product 5 (selina-4,11-diene) on the HP-5MS column but was separated from other sesquiterpenes on the HP-Innowax column. The mass spectrum of compound 14 (see Figure 3) closely resembled the mass spectrum of cadina-4,11-diene (Bohlmann et al 1984 (Bouwmeester et al 1999b) was later renamed amorpha-4,11-diene (Connolly and Hill 1991). Considering the structure and configuration, amorpha-4,11-diene is a suitable candidate for the olefinic first intermediate in the biosynthesis of artemisinin (Figure 1).…”

Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 86%

“…In leaf extracts of A. annua we detected 14 sesquiterpene olefins, of which ten could be identified using GC-MS libraries (Wiley and personal library of W.A. König), retention indices (Adams 1995) and authentic standards (see Figure 2) (Bouwmeester et al 1999b Compounds: 1,2,3,4,5,6,germacrene D;7,unknown 1;8,9,bicyclogermacrene;10,germacrene A;11,12,unknown 2;13,14, Redrawn from (Bouwmeester et al 1999b) presence of compound 14 was that we used two different GC-MS columns. Compound 14 is a minor compound that co-eluted with the major product 5 (selina-4,11-diene) on the HP-5MS column but was separated from other sesquiterpenes on the HP-Innowax column.…”

Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 99%

“…When a crude enzyme extract of the field-grown A. annua was assayed for sesquiterpene synthase activity with [ 3 H]-farnesyl diphosphate as a substrate, a range of labelled sesquiterpenes was detected by radio-GC (Bouwmeester et al 1999b). In order to be able to identify the individual sesquiterpenes (and hence their corresponding synthases) we used partial enzyme purification.…”

Section: Isolation and Characterization Of Amorpha-411-diene Synthasementioning

confidence: 99%

“…Using DEAE anion exchanger batch incubation, (NH 4 ) 2 SO 4 precipitation and Mono-Q anion-exchange chromatography (FPLC), amorpha-4,11-diene synthase was further purified. The enzyme preparation thus obtained was used to show that the enzyme had a K m of about 0.6 μM, a molecular mass of 56 kDa and a broad pH optimum around 6.5-7.0 (Bouwmeester et al 1999b). …”

Section: Isolation and Characterization Of Amorpha-411-diene Synthasementioning

confidence: 99%

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Research to Improve Artemisinin Production for use in the Preparation of Anti-Malarial Drugs

Bouwmeester

Bertea²,

Kraker³

et al.

Medicinal and Aromatic Plants

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Abstract. An important group of antimalarial drugs consists of the endoperoxide sesquiterpene lactone artemisinin and its semi-synthetically prepared derivatives. Because of the development of resistance against other malarial drugs, the demand for artemisinin has rapidly increased during the past decade. However, the supply of artemisinin is troublesome as neither total nor semi-synthesis are economically feasible and the only plant species known to produce artemisinin, Artemisia annua L., contains only low amounts of this compound. For a directed strategy to improve artemisinin production, one of the first requirements is to have insight into the regulation of its biosynthesis. Surprisingly, until some years ago hardly anything was known about the biosynthesis of artemisinin, particularly about the early enzymatic steps leading to (dihydro)artemisinic acid. To elucidate this important missing part in the pathway, we have analysed the terpenoids in A. annua leaves and the presence of sesquiterpene synthases, the enzyme class that should theoretically catalyse the first step in the formation of artemisinin. This led to the discovery of amorpha-4,11-diene synthase, which indeed catalyses the first step in artemisinin biosynthesis and for which the corresponding gene was cloned shortly afterwards. A subsequent search for oxidized derivatives of amorpha-4,11-diene -which could be postulated to be intermediates en route to artemisinin -revealed the presence of artemisinic alcohol, dihydroartemisinic alcohol, artemisinic aldehyde, dihydroartemisinic aldehyde and dihydroartemisinic acid. We also demonstrated the presence of the biosynthetic enzymes amorpha-4,11-diene hydroxylase, artemisinic alcohol dehydrogenase, artemisinic aldehyde reductase and dihydroartemisinic aldehyde dehydrogenase in A. annua. From these results, we conclude that the early steps in artemisinin biosynthesis involve: cyclization of farnesyl diphosphate by amorpha-4,11-diene synthase to amorpha-4,11-diene, hydroxylation of amorpha-4,11-diene to artemisinic alcohol, oxidation of artemisinic alcohol to artemisinic aldehyde, reduction of the C11-C13 double bond yielding dihydroartemisinic aldehyde, and oxidation of dihydroartemisinic aldehyde to dihydroartemisinic acid. During this biochemical research we have also managed to develop a trichome isolation protocol which has not only accelerated our biochemical work but also enables us to clone -in a very directed way -the genes that encode the enzymes responsible for artemisinin production. This should in the next few years result in several approaches to boost artemisinin production. 276H. BOUWMEESTER ET AL.

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Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 81%

Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 86%

Section: Identification Of Olefinic Sesquiterpenes In a Annua Leavesmentioning

confidence: 99%

Section: Isolation and Characterization Of Amorpha-411-diene Synthasementioning

confidence: 99%

Section: Isolation and Characterization Of Amorpha-411-diene Synthasementioning

confidence: 99%

See 3 more Smart Citations

Research to Improve Artemisinin Production for use in the Preparation of Anti-Malarial Drugs

Bouwmeester

Bertea²,

Kraker³

et al.

Medicinal and Aromatic Plants

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Antimalarial Agents

Casteel¹

2003

Burger's Medicinal Chemistry and Drug Discovery

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Malaria continues to be a major global health problem and seems to be increasing in some parts of the world. By way of an introduction, relevant aspects of the disease, including incidence and resistance, and of the parasite and its biochemistry and genetics are presented. The chemotherapeutic agents now available to prevent and treat malaria are discussed individually. Some of these drugs have been in use for decades or even centuries, such as quinine, chloroquine, and primaquine. Other agents are of more recent origin including mefloquine, halofantrine, atovaquone, and the artemisinins. Chloroquine had been the drug of first resort for many years. It is inexpensive, well‐tolerated, and extremely effective where parasites remain sensitive. But the development of resistance to chloroquine has emphasized the need for new agents and strategies to treat malaria. New information on the genetic basis of chloroquine resistance and on the overall mechanism of action of chloroquine holds promise for advances in therapy. Another important tactic in dealing with infection by resistant parasites is the use of drug combinations. The combination of pyrimethamine and sulfadoxine has been very successful, but resistance is developing. Newer, highly effective combinations include atovaquone with proguanil, lumefantrine with artemether, and chlorproguanil with dapsone. The artemisinin group of compounds has made a powerful positive impact on malaria chemotherapy although their use in monotherapy is not recommended. Details of the mechanism of action of these drugs are becoming clearer. Knowledge about action should assist in addressing issues of possible toxicity. Great progress has been made in sequencing the genome of Plasmodium falciparum . New molecular targets have been identified as a result and efforts are underway to develop antimalarial agents based on these targets. Researchers around the world are also exploring natural products for yet another lead in antimalarial chemotherapy. “If we take as our standard of importance the greatest harm to the greatest number, then there is no question that malaria is the most important of all infectious diseases 1 .” “Ah, poor heart! he is so shaked of a burning quotidian tertian, that it is most lamentable to behold [2].”

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Terpenes

and¹

2006

Plant Secondary Metabolites

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Amorpha-4,11-diene synthase catalyses the first probable step in artemisinin biosynthesis

Cited by 245 publications

References 35 publications

Research to Improve Artemisinin Production for use in the Preparation of Anti-Malarial Drugs

Research to Improve Artemisinin Production for use in the Preparation of Anti-Malarial Drugs

Antimalarial Agents

Terpenes

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