疏水层析介质的制备及对谷胱甘肽转硫酶的分离纯化

Preparation of Hydrophobic Interaction Chromatography Media and the Purification of Glutathione S-Transferase

通过向环氧氯丙烷活化的Sepharose CL-6B中引入苯基,制备了Phenyl-Sepha-rose CL-6B凝胶并优化了偶联条件,以该合成的层析介质分离纯化了猪肝中的谷胱甘肽转硫酶。结果表明,在NaOH浓度为0.25mol.L-1,苯酚加入量为0.5mL.g-1凝胶,60℃下反应4.5h的条件下,偶联效果最好,偶联率达到95.5%。用该疏水层析介质纯化猪肝中的谷胱甘肽转硫酶,活性回收率为54.54%,纯化倍数为2.6。

Sepharose CL-6B was activated with epichlorohydrin. Phenyl-Sepharose CL- 6B was synthesized in the process of coupling phenyl to the activated Sepharose CL-6B. The optimal conditions were studied. The Glutathione S-transferases（GSTs） from liver was purified using synthesized Phenyl-Sepharose CL-6B. The result showed that the maximum coupling rate was 95.5% with 0. 25 mol for 4. 5 h. The GSTs activity recovery was 54.5·L^-1 NaOH,0. 5 mL·g^-1 phenol at 60℃ and the purification fold was 2. 6.