title: Non-conventional induction strategies for production of subunit swine erysipelas vaccine antigen in rE. coli fed-batch cultures.
abstract: In spite of the large number of reports on fed-batch cultivation of E. coli, alternative cultivation/induction strategies remain to be more deeply exploited. Among these strategies, it could be mentioned the use of complex media with combination of different carbon sources, novel induction procedures and feed flow rate control matching the actual cell growth rate. Here, four different carbon source combinations (glucose, glycerol, glucose + glycerol and auto-induction) in batch media formulation were compared. A balanced combination of glucose and glycerol in a complex medium formulation led to: fast growth in the batch-phase; reduced plasmid instability by preventing early expression leakage; and protein volumetric productivity of 0.40 g.L(-1).h(-1). Alternative induction strategies were also investigated. A mixture of lactose and glycerol as supplementary medium fully induced a high biomass population, reaching a good balance between specific protein production (0.148 gprot.gDCW (-1)) and volumetric productivity (0.32 g.L(-1).h(-1)). The auto-induction protocol showed excellent results on specific protein production (0.158 gprot.gDCW (-1)) in simple batch cultivations. An automated feed control based on the on-line estimated growth rate was implemented, which allowed cells to grow at higher rates than those generally used to avoid metabolic overflow, without leading to acetate accumulation. Some of the protocols described here may provide a useful alternative to standard cultivation and recombinant protein production processes, depending on the performance index that is expected to be optimized. The protocols using glycerol as carbon source and induction by lactose feeding, or glycerol plus glucose in batch medium and induction by lactose pulse led to rSpaA production in the range of 6 g.L(-1), in short fed-batch processes (16 to 20 h) with low accumulation of undesired side metabolites.
authors: Chemical Engineering Department, Federal University of São Carlos, Rodovia Washington Luís, km 235, São Carlos, SP Brazil.
authors: Adilson José
authors: AJ
authors: da Silva
authors: Antônio Carlos Luperni
authors: AC
authors: Horta
authors: Ana Maria
authors: AM
authors: Velez
authors: Mônica Rosas C
authors: MR
authors: Iemma
authors: Cíntia Regina
authors: CR
authors: Sargo
authors: Raquel Lc
authors: RL
authors: Giordano
authors: Maria Teresa M
authors: MT
authors: Novo
authors: Roberto C
authors: RC
authors: Giordano
authors: Teresa Cristina
authors: TC
authors: Zangirolami
keywords: Auto-induction; Bioreactor cultivation; Erysipelothrix rhusiopathiae; Lactose feeding; SpaA antigen
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