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Plant Cell Rep ; 34(9): 1551-60, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25981048

RESUMO

KEY MESSAGE: 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.


Assuntos
Alquil e Aril Transferases/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Nicotiana/genética , Panax/enzimologia , Proteínas de Plantas/metabolismo , Sapogeninas/metabolismo , Ácido 2,4-Diclorofenoxiacético/farmacologia , Reatores Biológicos , Vias Biossintéticas/genética , Células Cultivadas , Cromatografia Gasosa-Espectrometria de Massas , Genes de Plantas , Ginsenosídeos/biossíntese , Ginsenosídeos/química , Ácido Mevalônico/metabolismo , Panax/efeitos dos fármacos , Panax/genética , Plantas Geneticamente Modificadas , Sapogeninas/química , Saponinas/metabolismo , Triterpenos/metabolismo
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