硅胶基质弱阳离子交换剂的合成及蛋清中溶菌酶的分离

Synthesis of Silica-Based Weak Cation Exchanger and Separation of Lysozyme From Egg White

用间接法合成了硅胶基质弱阳离子交换剂(XIDACE-WCX),详细研究了合成填料的色谱性能,并与商品柱(Shim-Pack WCX—1和Poly CATA)进行了比较。结果表明:合成柱比商品柱具有较高的分辨能力。通过考察合成填料的疏水性、蛋白质的保留值与等电点(pI)的关系、流动相的pH值和盐浓度对蛋白质保留行为的影响,论证了合成填料的离子交换特征。利用合成的色谱填料从蛋清中分离出纯度较高的溶菌酶(在排阻色谱上仅显示一条谱峰),酶在柱上的活性回收率为107％。

A silica-based weak cation exchanger (XIDACE-WCX) has been synthesized by indirect method. The chromatographic performance of the synthesized packing was studied in detail, and compared with that of commercial column (Shim-Pack WCX-1 and Poly CATA) by high performance liquid chromatography. The results showed that the synthesized column has higher resolution power than that of Shim-Pack WCX-l(Shimadzu) and Poly CATA (Poly LC) . The ion exchange characterization of the synthesized packing was demonstrated with the aid of examination concerning the hydrophobicity of the chromatographic column, the relationship between isoelectric point and retention of proteins and the effects of pH and salt concentration in mobile phase on retention behaviors of proteins. Lysozyme form egg white was separated by the synthesized chromatographic packing. The isolated enzyme has higher purity and appears a single-peak on size-exclusion chromatogram. The activity recovery of lysozyme is 107% through XIDACE-WCX column.