title: Production of dammarenediol-II triterpene in a cell suspension culture of transgenic tobacco.
abstract: Dammarenediol-II is biologically active tetracyclic triterpenoid, which is basic compound of ginsenoside saponin. Here, we established the dammarenediol-II production via a cell suspension culture of transgenic tobacco overexpressing PgDDS. Dammarenediol-II synthase catalyzes the cyclization of 2,3-oxidosqualene to dammarenediol-II, which is the basic triterpene skeleton in dammarene-type saponin (ginsenosides) in Panax ginseng. Dammarenediol-II is a useful candidate both for pharmacologically active triterpenes and as a defense compound in plants. Dammarenediol-II is present in the roots of P. ginseng in trace amounts because it is an intermediate product in triterpene biosynthesis. In this work, we established the production of dammarenediol-II via cell suspension culture of transgenic tobacco. The dammarenediol-II synthase gene (PgDDS) isolated from P. ginseng was introduced into the Nicotiana tobacum genome under the control of 35S promoter by Agrobacterium-mediated transformation. Accumulation of dammarenediol-II in transgenic tobacco plants occurred in an organ-specific manner (roots > stems > leaves > flower buds), and transgenic line 14 (T14) exhibited a high amount (157.8 μg g⁻¹ DW) of dammarenediol-II in the roots. Dammarenediol-II production in transgenic tobacco plants resulted in reduced phytosterol (β-sitosterol, campesterol, and stigmasterol) contents. A cell suspension culture was established as a shake flask culture of a callus derived from root segments of transgenic (T14) plants. The amount of dammarenediol-II production in the cell suspension reached 573 μg g⁻¹ dry weight after 3 weeks of culture, which is equivalent to a culture volume of 5.2 mg dammarenediol-II per liter. Conclusively, the production of dammarenediol-II in a cell suspension culture of transgenic tobacco can be applied to the large-scale production of this compound and utilized as a source of pharmacologically active medicinal materials.
authors.affiliation: Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chunchon, 200-701, Republic of Korea.
authors.forename: Jung-Yeon
authors.initials: JY
authors.lastname: Han
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Wang
authors.forename: Yong-Eui
authors.initials: YE
authors.lastname: Choi
mesh_terms: D000085:Acetates / Q000494:pharmacology; D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D003517:Cyclopentanes / Q000494:pharmacology; D035264:Flowers / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D036145:Ginsenosides / Q000096:biosynthesis; D060847:Metabolic Engineering; D009928:Organ Specificity; D054883:Oxylipins / Q000494:pharmacology; D005894:Panax / Q000201:enzymology* / Q000235:genetics; D010937:Plant Growth Regulators / Q000494:pharmacology; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012503:Saponins / Q000096:biosynthesis*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014315:Triterpenes
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D036145:Ginsenosides; D054883:Oxylipins; D010937:Plant Growth Regulators; D012503:Saponins; D014315:Triterpenes; C106188:dammarenediol; C072239:methyl jasmonate; D019883:Alkyl and Aryl Transferases; C514651:dammarenediol-II synthase, Panax ginseng