Production Of Cell Biomass And Artemisinin From Cell Cultures Of Artemisia Annua L. From Shake Flasks To Innovated Cell Culture Vessel
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Date
2014-07
Authors
Heng, King Wey
Journal Title
Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
Artemisia annua (Asteraceae) has been known to produce artemisinin, a potent antimalarial drug. Artemisinin is mainly produced in the leaves of field grown A. annua. Production of artemisinin has been hindered by low and inconsistent yields from field grown A. annua plants due to environmental and somatic variations, high labour costs involved in large scale planting and the complexity chemical synthesis of artemisinin. The limited supply of artemisinin has led to increasing market demand and soaring price of artemisinin as a first-line malarial drug to be used in artemisinin-based combination therapy (ACT). To achieve economical feasibility of artemisinin production, mass production of this compound can be done via plant cell culture technology. In this study, selected clones of A. annua (TC1 and TC2) callus cultures were used for the establishment of cell suspension cultures in shake flasks to optimized production of cell biomass and artemisinin content. Results obtained indicated that artemisinin production from A. annua cells decreased with the increasing culture volume employed, and the stirred culture vessel design was found to be superior for large volume production. The impeller rotational speeds and introduced aeration rates employed in the stirred culture vessel were found to have synergistic effects on the production of A. annua dried cell mass. The performance of the culture vessel was further optimized via numerical optimization to achieve 10.02 g/L and 5.91 g/L dried cell mass of A. annua TC1 clone and TC2 clone respectively, based on the Response Surface Methodology. Further studies indicated that impeller blade vertical slant of 30o was efficient in improving dried cell mass of A. annua TC1 and TC2 to 13.06 g/L and 6.26 g/L respectively. Addition of mevalonolactone 10 mg/L on day 8 into the cell culture of A. annua TC1 clone led to enhancement of 48.9 % artemisinin content corresponding to 552.1 ± 4.8 μg/g DW. The application of introduced aeration rates of 0.07 vvm and impeller rotational speeds of 103 rpm using an impeller blade of 30o vertical slant angle with the addition of mevalonolactone of 10 mg/L on day 8 in the stirred culture vessels proved that large culture volume of A. annua cells could achieve up to 13.13 ± 0.11 g/L dried cell mass and 596.8 ± 8.8 μg/g DW artemisinin content. This study hence indicated that large-scale culture of A. annua cells for artemisinin production could be viable, benefiting future industrial applications of cell culture for pharmaceutical drug production and ultimately increasing the availability and accessibility of these sought-after drugs (artemisinin) to the community.