| Full text | |
| Author(s): |
da Silva, Adilson Jose
[1]
;
Luperni Horta, Antonio Carlos
[1]
;
Velez, Ana Maria
[1]
;
Iemma, Monica Rosas C.
[1]
;
Sargo, Cintia Regina
[1]
;
Giordano, Raquel L. C.
[1]
;
Novo, Maria Teresa M.
[2]
;
Giordano, Roberto C.
[1]
;
Zangirolami, Teresa Cristina
[1]
Total Authors: 9
|
| Affiliation: | [1] Univ Fed Sao Carlos, Chem Engn Dept, BR-13560 Sao Carlos, SP - Brazil
[2] Univ Fed Sao Carlos, Genet & Evolut Dept, BR-13560 Sao Carlos, SP - Brazil
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | SPRINGERPLUS; v. 2, 2013. |
| Web of Science Citations: | 9 |
| 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 g(prot).g(DCW)(-1)) and volumetric productivity (0.32 g.L-1.h(-1)). The auto-induction protocol showed excellent results on specific protein production (0.158 g(prot).g(DCW)(-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. (AU) | |