应用Octyl Sepharose FF疏水层析分离好食脉孢霉发酵产生的纤溶酶

Separation of the fibrinolytic enzyme produced by N.sitophila by using Octyl Sepharose FF hydrophobic interaction chromatography

疏水层析是分离纯化蛋白质和多肽等生物大分子的常用方法,采用Octyl Se pharose FF疏水层析填料对好食脉孢霉(N.sitophila)发酵产生的纤溶酶进行了初步分离,比较了样品初始盐浓度、缓冲液pH值和洗脱方式对分离效果的影响。实验结果表明,采用样品初始盐浓度为40%、缓冲液pH值为7.4和步阶式梯度洗脱方式对样品分离效果较好,经测定样品中有3种疏水性不同的纤溶活性组分,其中活性组分Ⅲ经Octyl Sepharose FF疏水层析分离后比活力为855.56U/mg,总纯化倍数为7.12倍。

Hydrophobic interaction chromatography is a frequently-used method for purifying biomacromolecules such as polypeptide and protein. This article demonstrates the application of Octyl Sepharose FF hydrophobic interaction chromatography in purification of the fibrinolytic enzyme produced by N. sitophila. The purification effects of initial salinity, pH of buffer and elution mode were compared. The results showed that a better purified production can be received by adopting the steps-type gradient elution with the sample initial salinity of 40% and the buffer solution pH of 7.4. There are three kinds of hydrophobic fibrinolytic active components. One of them was called component III after being produced by N. sitophila. Octyl Sepharose FF hydrophobic interaction chromatography reached a specific activity of 855.56U/mg with a 7.12-fold purification.