RESUMO
The production of tropane alkaloids during differentiation and de-differentiation of Scopolia parviflora calli was studied. Tropane alkaloid production drastically decreased during calli de-differentiation. Scopolamine (1) production decreased after 10 days of culture, whereas that of hyoscyamine (2) decreased during de-differentiation of root to calli. The production of 1 was enhanced in calli undergoing differentiation to shoot after 60 days of culture, reaching a maximum by 80 days. However, production of hyoscyamine in regenerated plants was lower. The expression level of hyoscyamine 6beta-hydroxylase (H6H), a key biosynthetic enzyme for tropane alkaloids, was significantly increased in 4-week-old calli. This study suggests that the biosynthesis of tropane alkaloids is regulated inversely in de-differentiating Scopolia parviflora calli.
Assuntos
Escopolamina/biossíntese , Scopolia/química , Tropanos/isolamento & purificação , Coreia (Geográfico) , Estrutura Molecular , Raízes de Plantas/química , Scopolia/crescimento & desenvolvimento , Tropanos/químicaRESUMO
The effect of biotic elicitors on the production of bilobalide and ginkgolides in Ginkgo biloba cell suspension cultures was studied. The treatment of cell cultures with Candida albicans and Staphylococcus aureus as elicitors increased the amounts of bilobalide (BB), ginkgolide A (GA) and ginkgolide B (GB), with slight growth inhibition. The native bacterial elicitor was more effective for secondary metabolite accumulations both in cells and culture medium than autoclaved. However, exposure times of the cells to the elicitors strongly influenced the production of BB, GA and GB. This study suggests that biotic elicitors can regulate the production of BB, GA and GB either directly or indirectly. These results also describe the establishment of optimum conditions that determine the effects of biotic elicitors on secondary metabolism of bilobalides.
Assuntos
Candida albicans , Ciclopentanos/metabolismo , Furanos/metabolismo , Ginkgo biloba/crescimento & desenvolvimento , Ginkgolídeos/metabolismo , Staphylococcus aureus , Proliferação de Células , Células Cultivadas , Meios de Cultivo Condicionados , Interpretação Estatística de Dados , Ginkgo biloba/citologia , Ginkgo biloba/metabolismo , Sesquiterpenos , Terpenos/metabolismo , FitoalexinasRESUMO
The mass production of tropane alkaloids from adventitious root cultures of Scopolia parviflora, in small-scale bubble column bioreactor (BCB) was attempted. Adventitious roots of S. parviflora produced relatively enhanced levels of scopolamine and hyoscyamine in bioreactor compared to flask type cultures, and rapidly produced root clumps, with continuously increasing biomass throughout the culture period. The production of scopolamine and hyoscyamine in the top and bottom regions of root clumps were higher than in the core region. The adventitious root cultures of S. parviflora in the BCB required a relatively high level of aeration. The optimized conditions for the bioreactor culture growth and alkaloid production were found to be 3g of inoculum, on a fresh weight basis, a 15-day culture period and 0.4vvm of airflow. The elicitation by Staphylococus aureus increased the specific compound of scopolamine, while the production of hyoscyamine was slightly inhibited in BCB cultures.
Assuntos
Alcaloides/biossíntese , Reatores Biológicos , Raízes de Plantas/metabolismo , Scopolia/metabolismo , Tropanos/metabolismo , Atropina/biossíntese , Biomassa , Técnicas de Cultura de Células/métodos , Cinética , Escopolamina/biossínteseRESUMO
The effect of precursor feeding on the production of bilobalide and ginkgolides was studied with suspension cell cultures of Ginkgo biloba. The precursors greatly influenced the productivity of bilobalide and ginkgolides. Precursor supplementation increased the accumulation of both bilobalide and ginkgolides, and with positive effect on cell growth. The GA accumulation by cell cultures was influenced by precursors upstream in the metabolism, whereas the BB accumulation was under the influence of downstream precursors of the terpenoid biosynthetic pathway. Furthermore, precursor feeding modified the ratios of the BB, GA and GB in cells and cell cultures of G. biloba. The studies also aid in understanding effect of precursor feeding on the bilobalide and ginkgolides biosynthetic pathway.
Assuntos
Bilobalídeos/metabolismo , Técnicas de Cultura de Células/métodos , Meios de Cultivo Condicionados/metabolismo , Ginkgo biloba/efeitos dos fármacos , Ginkgo biloba/metabolismo , Ginkgolídeos/metabolismo , Terpenos/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Meios de Cultivo Condicionados/farmacologia , Terpenos/farmacologiaRESUMO
Cell suspension cultures of Capsicum annuum L. cv. P1482 were fed with exogenous ferulic acid to monitor their biotransformation abilities. A portion of the ferulic acid was biotransformed into vanillin, a major natural flavor, and capsaicin, a principle secondary metabolite characteristic of Capsicum species. The cellular vanillin concentrations were relatively higher than capsaicin levels and were maximal (2 mg/g DW) 4 days after 0.6 mM ferulic acid feeding. Maximal vanillin levels in the culture medium were 10 mg/L at 4 and 3 days after feeding with 1.25 and 2.5 mM ferulic acid, respectively. With regard to capsaicin levels, the cellular levels were slightly decreased by ferulic acid feeding, whereas the levels in the culture medium were increased. Ferulic acid feeding not only enhanced vanillin and capsaicin production but also increased the concentrations of other phenylpropanoid metabolites.