Study On The Fermentation Technology Of Recombinant Hirudin Expression In Pichia Pastoris

Natural hirudin is a kind of small molecule active peptide secreted from the salivary gland of Hirudo medicinals, is a most strong anticoagulin so far, can effectively cure thrombus disease. While the resource of natural hirudin is limited, low production, high production cost, thus restricting the application in clinic. The development of recombinant hirudin by using gene technology can hurdle the shortcom.The aim of the present study was to screening factors that determining the production of recombinant hirudin in Pichia pastoris. Effects of dissolved oxygen, temperature, pH, and additives on cell proliferation and on hirudin production were investigated in flask. And then factors were set appropriate levels to test the efficiency in 30 L bioreactor. The findings were (i) 30-50 % dissolved oxygen benefit cell growth and producing recombinant protein; (ii) temperature and pH should be set 24°C and 4.0, respectively; (iii) the production of hirudin should be triggered when cell wet reached 320 g/ml; (iv) the addition of 1 % casein hydrolysate increased the production of hirutin. The expression level was 28000 ATU/ml to 31000 ATU/ml.