Abstract
Recombinant-protein production in plants is a more effective system than the mammalian cell system because it can be more easily scaled up, has a low production cost, and is animal pathogen free. In this study, we established a brazzein production system using an air-lift bioreactor and transgenic carrot cells. The system included the introduction of the brazzein gene by Agrobacterium-mediated transformation. First, transgenic carrot callus (TC12) was cultured in a liquid medium. The brazzein content was analyzed during entire culture period. Next, the bioreactor type, volume, and culture period were compared to establish a bioreactor culture system. In addition, abscisic acid (ABA) was treated as an elicitor to increase brazzein biosynthesis. During the suspension culture, cell proliferation increased rapidly beginning at day 12. At that time, total soluble protein (TSP) and brazzein were 9.0 mg·g FW−1 and 1.2 mg·g FW−1, respectively. Brazzein content was 0.5 µg·g FW−1 (TSP 5.3 mg·g FW−1) in the balloon bioreactor. Cone and column bioreactors produced 1.1 µg·g FW−1 (TSP 5.7 mg·g FW−1) and 1.2 µg·g FW−1 (TSP 7.8 mg·g FW−1), respectively. The cone bioreactor was the most suitable for brazzein cell proliferation (6.1 g brazzein per bioreactor). After 8 h of 50 μM ABA treatment, the brazzein content had increased by 3.3-fold compared to that in the non-treated bioreactor. Overall, the results show that brazzein can be produced from carrot cell culture and an air-lift bioreactor system can be used for industrial mass production.
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Acknowledgements
This work was supported by a grant from the Next-Generation BioGreen21 Program (Project No. PJ01368901), and partially supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. NRF-2020R1A2C2102401). Authors are thanking to Ms. Yun-Ji Park who was technically helping to analysis of gene and protein expression, and Prof. Y.J. Jung (Hankyung National University) for providing brazzein vector.
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Han, JE., Lee, H., Ho, TT. et al. Brazzein protein production in transgenic carrot cells using air-lift bioreactor culture. Plant Biotechnol Rep 16, 161–171 (2022). https://doi.org/10.1007/s11816-022-00743-3
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DOI: https://doi.org/10.1007/s11816-022-00743-3