title: Growth kinetics and scale-up of Agrobacterium tumefaciens.
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 (μmax) of the strain in the three types of media was 0.46 ± 0.04 h-1in LB media, 0.43 ± 0.03 h-1in YEP media, and 0.27 ± 0.01 h-1in 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 kLa as the scale-up parameter.
authors.affiliation: Department of Chemical Engineering, University of California at Davis, Davis, CA, 95616, USA.
authors.forename: Ingrid K
authors.initials: IK
authors.lastname: Leth
authors.affiliation: Department of Chemical Engineering, University of California at Davis, Davis, CA, 95616, USA. kamcdonald@ucdavis.edu.
authors.forename: Karen A
authors.identifier: http://orcid.org/0000-0002-5145-9968
authors.initials: KA
authors.lastname: McDonald
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000254:growth & development*; D001431:Bacteriological Techniques; D018533:Biomass; D019149:Bioreactors; D003470:Culture Media / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D018515:Plant Leaves / Q000382:microbiology*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014026:Nicotiana / Q000033:anatomy & histology / Q000235:genetics / Q000382:microbiology
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins
keywords: Agrobacterium tumefaciens; Agroinfiltration; Recombinant proteins; Scale-up; Transient expression