毕赤酵母表达的人源Cu/Zn SOD的制备及稳定性分析

Preparation of recombinant human Cu/Zn SOD expressed in pichia pastoris and the study of its preservation condition

探讨了毕赤酵母表达人源Cu/Zn SOD的规模制备方法及稳定性研究.实验结果表明,采用45%乙醇复沉淀的方法来沉淀目标蛋白,目标蛋白回收率达88.89%,纯化倍数达到2.03倍,续而采用大孔树脂HPD500吸附的方法,去除了84.90%的色素,纯化倍数达4.60倍,比活为2700U·mg-1.经过纯化之后,将重组蛋白保存在4℃及低浓度、pH为7.8磷酸盐缓冲液条件37d,仍具有76.74%的活性.

Ethanol and macroporous resin was employed to establish a rapidly and efficient SOD industrial purification system, which was expressed by methylotrophic yeast, and then protein preservation conditions were tested by heating and salt testing. Datas showed that the recovery rate of SOD up to 88.89%, when fermentation liquid was treat by 45% ethanol for twice. Meanwhile, remained pigment was adsorpted by macroporous resin HPD500, which removed 84.90% pigment and purifieation fold was 4.60 with specific activity 2700 U · mg^-1 It＇ s obvious that when SOD was preserved by the condition of 4℃ and low concentration PBS（pH7.8）, protein activity can be lasted for 37 days, for its bioaetivity is still account for 76.74%. It is worth to point out that freeze - thaw would dramatie reduce the activity of SOD, only 60.00% activity can be preserved after twice freeze - thaw.