Recombinant protein expression in Pichia pastoris in a bioreactor via semi-continuous mode

Pichia pastoris has been successfully used as an efficient expression system for heterologous protein production. Herein, we report the development of a semi-continuous process to improve the production of recombinant proteins in P. pastoris by its cultivation in a BIONET’s F1 benchtop 3-liter bioreactor equipped with a continuous process module (CPM).Six repetitive batches were conducted in a semi-continuous process by replacing 2/3 of the culture every 72-96 h. The overall process in a 3-liter culture yielded a protein concentration of 0.28±0.13 mg·mL−1, taking 650 h to obtain 6±0.5 g of total target protein. On the contrary, the obtention of the equivalent amount of protein with a fed-batch mode would require around 1100 h of total time, including the inoculum preparation, the non-induced fermentation stage and the expression stage. Therefore, a reduction of the operation time and of the operation cost is evident when using a scalable semi-continuous fermentation strategy. These results suggest there is potential for the development of cost effective processes for the expression of recombinant proteins in P. pastoris in semi-continuous mode as an alternative to the productive continuous process mode, that is not available when low dilution rates are required due to the lack of available pumping technology.

Graph semi-continuous Pichia

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Recombinant protein expression in Pichia pastoris in a bioreactor via semi-continuous mode

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