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. 2018 Nov 10:20:e00292.
doi: 10.1016/j.btre.2018.e00292. eCollection 2018 Dec.

Double-stranded RNA production and the kinetics of recombinant Escherichia coli HT115 in fed-batch culture

Ljubomir Papić  1   2 , José Rivas  1 , Soledad Toledo  1 , Jaime Romero  1
Affiliations

Double-stranded RNA production and the kinetics of recombinant Escherichia coli HT115 in fed-batch culture

Ljubomir Papić et al. Biotechnol Rep (Amst). .

Erratum in

Abstract

Double-stranded RNA can induce interference processes. The specificity of this system raises the possibility of using dsRNA for therapeutic applications targeting viral diseases. Escherichia coli HT115 (DE3) has been widely used to produce dsRNA; however, the kinetics of dsRNA production and the relationship between dsRNA and biomass remain unknown. Our aims were to study the kinetics of dsRNA production and to improve dsRNA productivity with fed-batch technology. The results revealed that the production of dsRNA was growth-associated. In batch fermentation, the dsRNA/biomass yield remained close to 0.06 g·g-1, with a maximum productivity of 11.1 mg l-1 h-1 at 10 h of culture. In fed-batch fermentation, the yield was 0.06 g g-1, with a maximum dsRNA productivity of 15.2 mg l-1 h-1 at the end of the feed (12 h). Therefore, to increase the production of dsRNA, it is necessary to enhance the biomass that produces the recombinant nucleic acid.

Keywords: Escherichia coli HT115; Fed-batch; Luedeking-Piret kinetics; dsRNA production.

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Figures

Fig. 1
Fig. 1
Production of dsRNA in batch culture of E. coli HT115 (DE3) induced with lactose at 2nd hour of fermentation. a Bacterial growth curve in batch culture, the line is a model using Monod kinetics (μmax 0.27 h−1, K 0.1 g·l−1, r2 0.97). b dsRNA production in batch culture, the line is a growth-associated model using Luedeking-Piret kinetics (α 90, β 0, r2 0.96). Columns are dsRNA/biomass yield.
Fig. 2
Fig. 2
Agarose gel electrophoresis of dsRNA expressed in 12 h batch culture of E. coli HT115 (DE3) induced with lactose. Lane 2, 3 and 4 are triplicates. Arrow indicate dsRNA band that resolved at ∼480 bp.
Fig. 3
Fig. 3
Production of dsRNA in fed-batch culture of E. coli HT115 induced with lactose at 2nd hour of fermentation. Bacterial growth curve in fed-batch with exponential feeding, the line is a model using Monod kinetics (μmax 0.27 h−1, F0 0.0187 l·h−1, r2 0.95). Columns are dsRNA/biomass yield.
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