Functional Analysis of Amorpha-4,11-Diene Synthase (ADS) Homologs from Non-Artemisinin-Producing Artemisia Species: The Discovery of Novel Koidzumiol and (+)-α-Bisabolol Synthases.
The production of artemisinin, the most effective antimalarial compound, is limited to Artemisia annua. Enzymes involved in artemisinin biosynthesis include amorpha-4,11-diene synthase (ADS), amorpha-4,11-diene 12-monooxygenase (CYP71AV1) and artemisinic aldehyde Δ(11)13 reductase (DBR2). Although artemisinin and its specific intermediates are not detected in other Artemisia species, we reported previously that CYP71AV1 and DBR2 homologs were expressed in some non-artemisinin-producing Artemisia plants. These homologous enzymes showed similar functions to their counterparts in A. annua and can convert fed intermediates into the following products along the artemisinin biosynthesis in planta These findings suggested a partial artemisinin-producing ability in those species. In this study, we examined genes highly homologous to ADS, the first committed gene in the pathway, in 13 Artemisia species. We detected ADS homologs in A. absinthium, A. kurramensis and A. maritima. We analyzed the enzymatic functions of all of the ADS homologs after obtaining their cDNA. We found that the ADS homolog from A. absinthium exhibited novel activity in the cyclization of farnesyl pyrophosphate (FPP) to koidzumiol, a rare natural sesquiterpenoid. Those from A. kurramensis and A. maritima showed similar, but novel, activities in the cyclization of FPP to (+)-α-bisabolol. The unique functions of the novel sesquiterpene synthases highly homologous to ADS found in this study could provide insight into the molecular basis of the exceptional artemisinin-producing ability in A. annua.
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.
Paskorn
P
Muangphrom
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.
Hikaru
H
Seki
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan KNC Laboratories Co., Ltd., 3-2-34 Takatsukadai, Nishi-ku, Kobe, Hyogo, 651-2271 Japan.
Munenori
M
Suzuki
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan KNC Laboratories Co., Ltd., 3-2-34 Takatsukadai, Nishi-ku, Kobe, Hyogo, 651-2271 Japan.
Aya
A
Komori
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.
Mika
M
Nishiwaki
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.
Ryota
R
Mikawa
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan Continuing Professional Development Center, Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan.
Ery Odette
EO
Fukushima
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871 Japan muranaka@bio.eng.osaka-u.ac.jp.
Toshiya
T
Muranaka
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000962:Antimalarials / Q000737:chemistry / Q000378:metabolism; D019306:Artemisia / Q000201:enzymology* / Q000235:genetics; D031172:Artemisia annua / Q000201:enzymology / Q000235:genetics; D037621:Artemisinins / Q000737:chemistry / Q000378:metabolism; D053898:Biosynthetic Pathways; D007887:Fabaceae / Q000201:enzymology* / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D007783:Lactones / Q000737:chemistry / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D000081222:Monocyclic Sesquiterpenes; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism
D000962:Antimalarials; D037621:Artemisinins; D007783:Lactones; D000081222:Monocyclic Sesquiterpenes; D010940:Plant Proteins; D000081223:Polycyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C515348:amorpha-4,11-diene; C004497:bisabolol; C004808:farnesyl pyrophosphate; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C531287:artemisinic aldehyde delta11(13) reductase, Artemisia annua; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase; C075875:artemisin
(+)-α-Bisabolol; Amorpha-4,11-diene synthase; Artemisia absinthium; Artemisia kurramensis; Artemisia maritima; Koidzumiol