| 摘要 |
PURPOSE:To purify an extracellular aminopeptidase by four chromatographic treatments and two dialysis treatments. CONSTITUTION:Cultured liquid of Penicillium caseicolum is centrifuged and filtered to obtain a purified enzyme liquid, which is salted out with saturated ammonium sultate. The supernatant liquid is dialyzed with 0.02M phosphoric acid buffer solution and then concentrated. The concentrated solution is subjected to the following three DEAE-sepharose chromatography treatments. I: Elution with 0-0.3M KCl by concentration gradient method using a 2.0X40cm column, a flow rate of 30ml/hr, a fraction of 5ml and 0.02M phosphoric acid buffer solution (6.5pH); II: elution with 0.13-0.14M KCl using a 1.5X30cm column, a flow rate of 20ml/hr and a fraction of 5ml and III: elution with 0.15M KCl using a 1.0X20cm column and a flow rate of 15ml/hr. The eluted liquid is dialyzed with 0.02M phosphoric acid buffer solution and concentrated. Finally, the concentrated liquid is eluted by chromatofocusing process of PBE94 gel using a polybuffer with a 1.0X15cm column, a flow rate of 15ml/hr and a fraction of 2ml. |
