Large-scale fermentation and purification of rhHSP110 in Pichia pastoris

Objective To study large-scale fermentation of recombinant human HSP110(rhHSP110) in Pichia pastoris,and the purification of expressed target protein in Pichia pastoris.Methods rhHSP110(2 L) was prepared in shake flask.Using fed-batch fermentation,the high-density fermentation of genetically engineered Pichia pastoris was processed in a 80 L bioreactor.After induction for 72 h,the target protein was purified by SP Sepharose XL.Results The fermentation temperature was set at 30℃,the pH value was 4.2,and the DO was kept over 20%.When the wet weight of the cells reached 200 g·L-1,the methanol-induced phase was initiated.By cation exchange chromatography it was possible to purify the rhHSP110 produced in Pichia pastoris,and a yield of about 500 mg·L-1 was reached.Conclusion The success of large-scale fermentation lays a foundation for mass production and clinical applications of rhHSP110.