Bioreactors provide suitable conditions for the growth of cells and production of secondary metabolites by regulating physical and chemical factors. In this study, rst, sucrose, 2-(N-morpholino) ethanesulfonic acid (MES) as a buffering agent and medium pH was optimized in the Erlenmeyer ask. This aim was then pursued in a stirred bioreactor through aeration and pH medium adjustment. Results of the rst step showed that Schenk and Hildebrandt (SH) basal medium with naphthalene acetic acid (2 mg.l -1 ) and 6benzylaminopurine (1 mg.l -1 ) supplemented with 2.5 mM of MES and gradually increment of sucrose from 3 to 6% caused to catch the highest cell biomass and crocin production. The spectrophotometry measurement showed that the highest crocin content of the cells was 0.8 mg/g after ve weeks. The results of the second part revealed that in the stirred bioreactor, constant pH (5.8) during the growth period is a limited factor for the cell growth and crocin production. Although aeration initially found to be an inhibited factor for the production of crocin, results showed that, if the evaporated volume of water caused by aeration is constituted, it can be an effective factor to increase cell growth rate around 2 folds.In addition, total crocin content of the cells, based on the HPLC could be raised up to 2 mg/g. Based on this study, it can be concluded that MES and gradual increment of sucrose could increasing the cell growth and crocin production. Aeration in bioreactor can increase cell biomass, if the medium volume will be kept constant.
Key MessageSaffron, the most expensive spice contains valuable compounds like crocin. Corm derived-cell containing crocin can be produced in higher scales and cheaper price by using cell culture in stirred bioreactor.
The cell suspension culture and seed samples of Bunium persicum were extracted by supercritical fluid, hydrodistillation and solvent methods and analyzed by Gas Chromatography. In this study to compare the different methods of extractions, cuminaldehyde was targeted as one of the Black zira essential oil constitute. For callus induction the germinated seeds were cultured as explants on Murashige and Skoog medium supplemented with 2 mg/l 2,4-dichlorophenoxy acetic acid and 0.5 mg/l kinetin (treatment A) as well as 2 mg/l α-naphthalene acetic acid and 0.5 mg/l 6-benzyl aminopurine (treatment B) and followed by cells suspension cultures establishment for the first time. The results of cell culture showed that cells from treatment B have a growth rate higher than A. All extracts were dissolved in 1 ml hexane and analyzed by Gas Chromatography. According to the Gas Chromatography analysis, cuminaldehyde was not detected in the supercritical fluid samples, while it was present in hydrodistillation and solvent extract. Cuminaldehyde percentage in cell and seed solvent extracts was 4.65% and 18.61% respectively. Gas Chromatography results also showed that no cuminaldehyde is present in media extracts, means no cuminaldehyde has been secreted into the medium.
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