FERMENTATION TECHNIQUE WITH PICHIA YEAST EXPRESSING RECOMBINANT PROTEIN

The present invention discloses a fermentation process with a Pichia yeast expressing a recombinant protein, in which Pichia pastoris is used as a fungal strain. The process comprises performing primary seed culturing to reach a fungal concentration of 20±2 g/L; then performing secondary seed culturing to reach a fungal concentration of 120±10 g/L; next proceeding to a glycerol culturing stage, wherein the amount of glycerol added in the glycerol culturing stage is 60-70 g/L; and then proceeding to a methanol-induced stage for 120±8 h after the dissolved oxygen quickly reaches a relatively stable state, to complete the fermentation process. In the present invention, a glycerol fed-batch addition stage in existing processes is omitted, and the process proceeds to a next stage as soon as glycerol is completely consumed, with no need to prepare sterilized glycerol for fluidic addition. As such, the glycerin sterilizer is omitted, the consumption of energy and resource and the waste of glycerin are reduced. Moreover, the fungal concentration has no need to be monitored, thus reducing the probability of errors, and lowering the fermentation failure caused by technical errors. Therefore, the present process is more suitable for large-scale industrial production, and brings convenience and great economic value for the industrial production of the recombinant protein.