title: Production of the dammarene sapogenin (protopanaxadiol) in transgenic tobacco plants and cultured cells by heterologous expression of PgDDS and CYP716A47.
abstract: Protopanaxadiol (PPD) is an aglycone of dammarene-type ginsenoside and has high medicinal values. In this work, we reported the PPD production in transgenic tobacco co-overexpressing PgDDS and CYP716A47. PPD is an aglycone of ginsenosides produced by Panax species and has a wide range of pharmacological activities. PPD is synthesized via the hydroxylation of dammarenediol-II (DD) by CYP716A47 enzyme. Here, we established a PPD production system via cell suspension culture of transgenic tobacco co-overexpressing the genes for PgDDS and CYP716A47. The concentration of PPD in transgenic tobacco leaves was 2.3-5.7 µg/g dry weight (DW), depending on the transgenic line. Leaf segments were cultured on medium with various types of hormones to induce callus. Auxin treatment, particularly 2,4-D, strongly enhanced the production of DD (783.8 µg g(-1) DW) and PPD (125.9 µg g(-1) DW). Treatment with 2,4-D enhanced the transcription of the HMG-Co reductase (HMGR) and squalene epoxidase genes. PPD production reached 166.9 and 980.9 µg g(-1) DW in a 250-ml shake flask culture and in 5-l airlift bioreactor culture, respectively.
authors.affiliation: Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chunchon, 200-701, Republic of Korea.
authors.forename: Ju-Hyeon
authors.initials: JH
authors.lastname: Chun
authors.forename: Prakash Babu
authors.initials: PB
authors.lastname: Adhikari
authors.forename: Seong-Bum
authors.initials: SB
authors.lastname: Park
authors.forename: Jung-Yeon
authors.initials: JY
authors.lastname: Han
authors.forename: Yong-Eui
authors.initials: YE
authors.lastname: Choi
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D019883:Alkyl and Aryl Transferases / Q000378:metabolism*; D019149:Bioreactors; D053898:Biosynthetic Pathways / Q000235:genetics; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D017343:Genes, Plant; D036145:Ginsenosides / Q000096:biosynthesis / Q000737:chemistry; D008798:Mevalonic Acid / Q000378:metabolism; D005894:Panax / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D010940:Plant Proteins / Q000378:metabolism*; D030821:Plants, Genetically Modified; D012502:Sapogenins / Q000737:chemistry / Q000378:metabolism*; D012503:Saponins / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics*; D014315:Triterpenes / Q000378:metabolism
chemical_list: D036145:Ginsenosides; D010940:Plant Proteins; D012502:Sapogenins; D012503:Saponins; D014315:Triterpenes; C106188:dammarenediol; D015084:2,4-Dichlorophenoxyacetic Acid; D003577:Cytochrome P-450 Enzyme System; D019883:Alkyl and Aryl Transferases; C514651:dammarenediol-II synthase, Panax ginseng; C062916:protopanaxadiol; D008798:Mevalonic Acid