柱层析法分离纯化菊粉酶的研究  

Column Chromatography Purification of Inulinase

作　　者：倪晓颖[1] 李秉超[2] 林荣峰[3] 李丽梅[2] 

机构地区：[1]沈阳农业大学食品学院,沈阳110866 [2]沈阳农业大学理学院,沈阳110866 [3]沈阳农业大学畜牧兽医学院,沈阳110866

出　　处：《沈阳农业大学学报》2010年第4期484-487,共4页Journal of Shenyang Agricultural University

摘　　要：以黑曲霉固体发酵法产生的粗菊粉酶为试材,采用葡聚糖凝胶层析法和DEAE纤维素52柱层析法对经过硫酸铵沉淀后的粗酶进行了分离纯化,得到接近电泳纯的高纯度菊粉酶。结果表明:以菊粉为底物研究酶活时,DEAE-纤维素52离子交换层析得到较好的分离效果,主要活力峰比活力12.64U·mg-1提纯了6.37倍;以蔗糖为底物时,SephadexG100分离效果好,比活力35.57U·mg-1提纯了3.78倍。With the inulinase produced by solid fermentation of Aspergillus niger as test material,crude enzyme was purified after ammonium sulfate precipitated by Sephadex chromatography and DEAE-cellulose 52 column chromatography,and then high purity inulinase close to electrophoretic purity was abtained.The results showed that when inulin was as the substrate of activity,DEAE-cellulose 52 ion exchange chromatography achieved good separation,and the main activity peak specific activity was 12.64 U·mg-1 with 6.37-fold purification;when sucrose was as the substrate,SephadexG100 achieved good separation,and purification of specific activity was 35.57 U·mg-1 with 3.78-fold purification.

关 键 词：菊粉酶 Sephadex凝胶层析 DEAE-纤维素离子交换层析 

分 类 号：Q533[生物学—生物化学]