Abstract
Production of recombinant proteins in plants through Agrobacterium-mediated transient expression is a promising method of producing human therapeutic proteins, vaccines, and commercial enzymes. This process has been shown to be viable at a large scale and involves growing large quantities of wild-type plants and infiltrating the leaf tissue with a suspension of Agrobacterium tumefaciens bearing the genes of interest. This study examined one of the steps in this process that had not yet been optimized: the scale-up of Agrobacterium production to sufficient volumes for large-scale plant infiltration. Production of Agrobacterium strain C58C1 pTFS40 was scaled up from shake flasks (50–100 mL) to benchtop (5 L) scale with three types of media: Lysogeny broth (LB), yeast extract peptone (YEP) media, and a sucrose-based defined media. The maximum specific growth rate (μ max) of the strain in the three types of media was 0.46 ± 0.04 h−1 in LB media, 0.43 ± 0.03 h−1 in YEP media, and 0.27 ± 0.01 h−1 in defined media. The maximum biomass concentration reached at this scale was 2.0 ± 0.1, 2.8 ± 0.1, and 2.6 ± 0.1 g dry cell weight (DCW)/L for the three media types. Production was successfully scaled up to a 100-L working volume reactor with YEP media, using k L a as the scale-up parameter.
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Acknowledgements
This work was supported by the National Science Foundation (CBET-1067423) and the National Science Foundation G-K12 RESOURCE Fellowship program (DGE-0948021) and received funding for equipment from Chevron Corporation (grant no. 0700679S024).
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Leth, I.K., McDonald, K.A. Growth kinetics and scale-up of Agrobacterium tumefaciens . Appl Microbiol Biotechnol 101, 4895–4903 (2017). https://doi.org/10.1007/s00253-017-8241-5
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DOI: https://doi.org/10.1007/s00253-017-8241-5