Effect of Agitation on Taq DNA Polymerase Production by Escherichia Coli in Bioreactor
Morales-Fonseca, Diana
Coronado-Jimenez, Laura
Gonzalez-Moya, Valentina
Zambrano, Maria Mercedes
Sandoval-Herrera, Juan
Arturo-Calvache, Jaime E.

How to Cite

Morales-Fonseca D., Coronado-Jimenez L., Gonzalez-Moya V., Zambrano M.M., Sandoval-Herrera J., Arturo-Calvache J.E., 2022, Effect of Agitation on Taq DNA Polymerase Production by Escherichia Coli in Bioreactor, Chemical Engineering Transactions, 93, 121-126.


The Taq DNA polymerase is an important protein used during PCR (Polymerase chain reaction), a technique widely used today. The production of this enzyme in bioreactors make possible to improve efficiency, to reduce production time, to increase production and expression, or to reduce production costs. This is achieved thanks to the measurement, monitoring and control of the different parameters that influence this biotechnological process: dissolved oxygen, pH, temperature, incubation time, agitation during the incubation and induction phase.In the present work, the production of Taq polymerase expressed in Escherichia coli, was made on a bioreactor scale, BioFlo/CelliGen 115®, with a volume of 1 L, equipped with temperature (37 °C) and pH (6.5) control, and with a constant level of dissolved oxygen provided by a 1 vvm flow of air. After the induction phase, we extracted the polymerase by cell disruption using centrifugation and some substances. For the protein measurement was implemented d technique and SDS-PAGE electrophoresis.
According to gel electrophoresis results, molecular weight of the protein obtained in bioreactor was 80 kD, similar to the value of commercial production in flasks. The effect of agitation on protein concentration after induction was studied at two levels: 220 and 350 rpm, the study found a production of 1465 ???? ?? -1 and 2975 ???? ?? -1 of protein respectively. The results confirm a higher production of protein in this bioreactor, keeping the type of polymerase. This could be the first step to scale the production with controlled conditions at higher rates.