Abstract
Bioreactors provide suitable conditions for the growth of cells and production of secondary metabolites by regulating physical and chemical factors. In this study, saffron cell growth and the production of crocin metabolite were evaluated in Erlenmeyer flask and in a stirred bioreactor. To optimize the conditions, the effect of different concentrations of 2-(N-morpholino) ethanesulfonic acid (MES) (0, 2.5 and 5 mM) as a buffering agent and sucrose (3%, 6% and gradually 6%) were addressed on saffron cell growth and crocin production in the Erlenmeyer flask. This aim was then pursued in a stirred bioreactor through investigation on the effect of aeration and pH medium adjustment. The results of the first step experiment showed that Schenk and Hildebrandt basal medium with naphthalene acetic acid (2 mg l−1) and 6-benzylaminopurine (1 mg l−1) supplemented with 2.5 mM of MES as well as gradual increment of sucrose from 3 to 6% caused the highest cell biomass and crocin production. The spectrophotometry measurement showed that the highest crocin content of the cells was 0.8 mg g−1 after 6 weeks. The results of the second part revealed that in the stirred bioreactor, constant pH (5.8) during the growth period is a limiting factor for the cell growth and crocin production. Aeration was also found to be an inhibiting factor for the production of crocin. The results showed that, if the evaporated volume of water caused by aeration is constituted, it can be an effective factor to increase cell growth around 2 folds. In addition, total crocin content of the proliferated cells in stirred bioreactor system under the non-constant pH and without aeration conditions, could be raised up to 2 mg g−1 of cell dry weight, based on the HPLC determination. According to the results of this study, it can be concluded that MES and gradual increment of sucrose could increase the cell growth and crocin production. In the tested stirred bioreactor, it was found that the natural pH fluctuation is a suitable condition for saffron biomass cell growth and crocin production. The proper aeration could increase the cell biomass, if the evaporated water is replaced. However based on the obtained results it can be concluded that, aeration at tested value is not capable of increasing the crocin production.
Key message
Saffron, 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.
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Data availability
The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.
Code availability
Not applicable.
Notes
Volume per Volume Medium.
Abbreviations
- BA:
-
6-Benzylaminopurine
- MES:
-
2-(N-morpholino) ethanesulfonic acid
- NAA:
-
Naphthalene acetic acid
- SH:
-
Schenk and Hildebrandt (1972)
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The authors would like to acknowledge from the Research Institute of Food Science and Technology (RIFST) for providing all lab facilities and Iran National Science Foundation (INSF) for financial supports of this project.
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The funding was provided by Research Institute of Food Science and Technology (RIFST), Iran.
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In this experiment, Ziaratnia and Hemmati designed the experiments. Amini carried out all the experiments, analyzed data and wrote the paper. All authors read and approved the manuscript.
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Amini, S., Ziaratnia, S.M. & Hemmati, K. Optimization of conditions for increasing of saffron cell biomass and crocin production in stirred bioreactor. Plant Cell Tiss Organ Cult 149, 243–255 (2022). https://doi.org/10.1007/s11240-022-02233-4
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DOI: https://doi.org/10.1007/s11240-022-02233-4