Control of Proteolysis of Recombinant Proteins in Escherichia coli
Year: 2001
Type: article
Abstract: Proteolysis was shown to have a significant impact on the accumulation and the final yield of recombinant proteins. Much smaller intracellular concentrations of the proteins SpA and ZZT2 compared to their stable versions SpA-βgalactosidase and ZZT0 were explained by their high proteolysis rate. The reduction of proteolysis rate in Clp protease-deficient strain enabled to increase the intracellular ZZT2 concentration two-fold, but impaired the cell growth. A method to calculate a hypothetical accumulation of the product assuming complete stabilization was described, which reveals a potential to increase the productivity by eliminating proteolysis. The effects of scale-up on the production of the ZZT2 protein were investigated in reactors of different configuration and scale: a well-mixed lab-scale bioreactor, an industrial scale 12-m3 bioreactor, and a two-compartment scale-down bioreactor. The production of the recombinant protein ZZT2 was highest in the lab-scale cultivation (50 mg/g), however in the large-scale the ZZT2 concentration declined 8 hours after induction from its highest value of 45 to a final value of 35 mg/g. The final yield of the ZZT2 protein in the scale-down
Author: Aleksei Rozkov
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W374634289