摘要
疏水层析是一种有效的分离纯化手段,本文采用Octyl Sepharose和Butyl Sepharose两种疏水层析填料对里氏木霉表达的t-PA的提纯操作条件进行了初步研究。结果表明,选用Butyl Sepharose FF填料,上样量10mL(蛋白含量4.1mg/mL),用15%(NH4)2SO4的PBS以2mL/min的流速进行洗脱时,回收的酶的比活性较大,为2959.54U/mg,纯化倍数为5.16,回收率为7.12%。
Hydrophobic interaction chromatography is an effective method for isolation and purification.This article demonstrates the application of Hydrophobic interaction chromatography to the purification of t-PA produced by Trichoderma reesei and the optimization of separating and purificating conditions of t-PA. The purification operation conditions of Octyl Sepharose FF and Butyl Sepharose FF columns wrer primarily researched. The result showed that Butyl Sepharose FF was suitable for separation of t-PA. The optimized chromatography condition was start elution butter 0.02mul/L PBS(pH7.4) contained 15% (NH4)2SO4 at a rate of 2mL/min. The enzyme specific activity is 2959.54U/mg.The purification multiple is 5.16.The t-PA activity recovery is 7.12%.
出处
《齐齐哈尔大学学报（自然科学版）》
2005年第1期15-18,共4页
Journal of Qiqihar University(Natural Science Edition)
关键词
里氏木霉
T-PA
疏水层析
Trichoderma reesei
t-PA
hydrophobic interaction chromatography
