The inhibition of aggregation of recombinant human consensus interferon-alpha mutant during Pichia pastoris fermentation.
Lower induction temperature and polyoxyethylene sorbitan monolaurate (Tween-20) were successfully used to inhibit the aggregation of recombinant human consensus interferon-alpha mutant (cIFN) during Pichia pastoris fermentation. When the induction temperature was decreased from 30 to 20 degrees C, the cIFN secreted into the medium was in the form of monomers instead of aggregates. The maximum specific activity at 20 degrees C was 4.04 times as high as that at 30 degrees C. There was no obvious effect on the cell growth at 20 degrees C, but the total protein level was decreased. Similar inhibition effect on cIFN aggregation was observed when 0.2 g l(-1) Tween-20 was added during induction. Furthermore, there was a synergistic effect found between induction temperature and Tween-20 on the inhibition of cIFN aggregation. The maximum specific activity with Tween-20 at 20 degrees C was 19.9-fold higher than that without Tween-20 at 30 degrees C.
Hao
Yuyou
Y
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P.O. box 329, Shanghai. People's Republic of China.
Chu
Ju
J
Wang
Yonghong
Y
Zhuang
Yingping
Y
Zhang
Siliang
S
D003902:Detergents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D007370:Interferon Type I / Q000235:genetics / Q000378:metabolism*; D016898:Interferon-alpha; D009154:Mutation; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011136:Polysorbates / Q000494:pharmacology*; D011994:Recombinant Proteins; D013696:Temperature
D003902:Detergents; D007370:Interferon Type I; D016898:Interferon-alpha; D011136:Polysorbates; D011994:Recombinant Proteins; C050739:interferon alfacon-1