Abstract
Plant-based recombinant protein production systems are powerful tools that can be used instead of microbial and animal cell culture systems. The aim of this study was to establish a stable brazzein production system from carrot cell suspension using bioreactor. Using Agrobacterium-mediated transformation (stress-inducible SWPA2 promoter and brazzein gene), three transgenic lines (TC1, 11, 12) were generated. During the TC12 culture period, cell proliferation increased rapidly up to 15 days, and the maximum cell division rate (log phase) was reached after 6 days of culture. Fresh weight peaked at day 18, 3.8-fold (7.6 g 100 mL−1) higher than on day 1, and gene expression was tenfold higher on day 27 than on day 0. The brazzein-encoding gene was also analyzed after treatment with various stress factors for increase of brazzein gene expression, and abscisic acid (ABA) and hydrogen peroxide (H2O2) proved most effective. In particular, the gene expression was enhanced 2.5-fold with 220 μM H2O2 and 2.8-fold with 50 μM ABA, compared with controls. Transgenic cells were then transferred to various types of air-lift bioreactor, and the column bioreactor achieved a higher biomass (238.9 g L−1) than cone and balloon bioreactors. These results demonstrate an efficient protein production system for brazzein using an air-lift bioreactor that may be suitable for the use in the food industry.
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Acknowledgements
This work was supported by a grant from the Next Generation BioGreen21 Program (Project No. PJ013689), Rural Development Administration, Republic of Korea, and authors thank to Dr. Sang-Soo Kwak of Korea Research Institute of Bioscience and Biotechnology KRIBB for providing SWPA promoter for the study.
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JE Han and YJ Park acquired the data and wrote the manuscript. H Lee and YJ Jeong participated in the interpretation of data and revision for important intellectual content. SY Park conceptualized and designed this study.
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Han, JE., Park, YJ., Lee, H. et al. Increased brazzein expression by abiotic stress and bioreactor culture system for the production of sweet protein, brazzein. Plant Biotechnol Rep 14, 459–466 (2020). https://doi.org/10.1007/s11816-020-00625-6
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DOI: https://doi.org/10.1007/s11816-020-00625-6