Metabolic engineering of Nicotiana benthamiana for the increased production of taxadiene.
We report the production of taxadiene by transformation of N. benthamiana with a taxadiene synthase gene. The production was significantly increased by an elicitor treatment or metabolic pathway shunting. Paclitaxel (Taxol(®)) was first isolated from the bark of the pacific yew tree as an anticancer agent and has been used extensively to treat various types of cancer. Taxadiene, the first committed product of paclitaxel synthesis is cyclized from geranylgeranyl diphosphate (GGPP), and further complex hydroxylation and acylation processes of the unique taxane core skeleton produce paclitaxel. To accomplish de novo production of taxadiene, we transformed Nicotiana benthamiana with a taxadiene synthase (TS) gene. The introduced TS gene under the transcriptional control of the CaMV 35S promoter was constitutively expressed in N. benthamiana, and the de novo production of taxadiene was confirmed by mass spectroscopy profiling. Transformed N. benthamiana homozygous lines produced 11-27 μg taxadiene/g of dry weight. The highest taxadiene production line TSS-8 was further treated with an elicitor, methyl jasmonate, and metabolic pathway shunting by suppression of the phytoene synthase gene expression which resulted in accumulation of increased taxadiene accumulation by 1.4- or 1.9-fold, respectively. In summary, we report that the production of taxadiene in N. benthamiana was possible by the ectopic expression of the TS gene, and higher accumulation of taxadiene could be achieved by elicitor treatment or metabolic pathway shunting of the terpenoid pathway.
School of Biotechnology, Yeungnam University, Gyeongbuk, Gyeongsan, 712-749, Korea.
Md Mohidul
MM
Hasan
Hyun-Soon
HS
Kim
Jae-Heung
JH
Jeon
Sung Hong
SH
Kim
BoKyung
B
Moon
Jai-Young
JY
Song
Sang Hee
SH
Shim
Kwang-Hyun
KH
Baek
D000085:Acetates / Q000494:pharmacology; D000475:Alkenes / Q000737:chemistry / Q000378:metabolism*; D000972:Antineoplastic Agents, Phytogenic / Q000096:biosynthesis / Q000737:chemistry; D001952:Bridged-Ring Compounds / Q000378:metabolism; D003517:Cyclopentanes / Q000494:pharmacology; D004224:Diterpenes / Q000737:chemistry / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D007535:Isomerases / Q000235:genetics* / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D054883:Oxylipins / Q000494:pharmacology; D017239:Paclitaxel / Q000096:biosynthesis / Q000737:chemistry; D010937:Plant Growth Regulators / Q000494:pharmacology; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis / Q000737:chemistry; D043823:Taxoids / Q000378:metabolism; D020946:Taxus / Q000201:enzymology* / Q000235:genetics; D014026:Nicotiana / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*
D000085:Acetates; D000475:Alkenes; D000972:Antineoplastic Agents, Phytogenic; D001952:Bridged-Ring Compounds; D003517:Cyclopentanes; D004224:Diterpenes; D054883:Oxylipins; D010937:Plant Growth Regulators; D011106:Polyisoprenyl Phosphates; D043823:Taxoids; C093125:taxa-4(5),11(12)diene; C080625:taxane; C072239:methyl jasmonate; D007535:Isomerases; C095926:taxa-4(5),11(12)-diene synthase; C002963:geranylgeranyl pyrophosphate; D017239:Paclitaxel