高效阳离子交换色谱法分离纯化蛋清中的溶菌酶交换色谱法分离纯化蛋清中的溶菌酶

李　蓉, 　陈国亮

(西北大学化工系, 陕西西安710069)

摘要: 建立了用高效阳离子交换色谱分离纯化蛋清中溶菌酶的新方法,讨论了纯化的工艺条件。蛋清样品匀浆后,用氯化钠初步纯化,然后用弱阳离子交换柱XIDACE2WCX分离。结果表明,被纯化的溶菌酶和杂蛋白得到很好分离。经活性检测,溶菌酶过柱后的活性回收率为107 % ,蛋白的比活为15 467 U/ mg ,纯化倍数提高了516 倍。用尺寸排阻色谱(SEC) 鉴定,得到的溶菌酶呈均一性。该法较传统软基质低压离子交换色谱分离速度快,纯化效率高。

关键词:高效阳离子交换色谱;溶菌酶;蛋清

中图分类号:O658 ;Q51 　　　文献标识码:A 　　　文章编号:100028713 (2002) 0320259203

Separation and Purif ication of Lysozyme from Egg White by High Performance Cation2Exchange Chromatography

L I Rong , CHEN Guo2liang

( Department of Chemical Engineering , Northwest University , Xi’an 710069 , China)

Abstract : A new method used to separate and purify lysozyme f rom egg white by high performance cation2exchange chromatography has been established. The process conditions for purifying lysozyme were also discussed in detail. The procedure involved that homogenization of the egg white sample ,preliminary purification with sodium chloride , and chromatographic separation by the weak cation ex2change column (XIDACE2WCX) . The experimental result s showed that the purified lysozyme and other impurity proteins were completely separated. By using bioactivity assay , the recovery of lysozyme was 107 % , and the specific activity was 15 467 U/ mg through the column. It s purity was raised 5162fold. The collected f raction with activity was detected by size2exclusion chromatography (SEC) . The purified lysozyme was homogeneous. Compared with the t raditional soft2based low pres2 sure ion2exchange chromatography , the developed method is rapid and effective.

Key words : high performance cation2exchange chromatography ; lysozyme ; egg white

正文：/uploadfile/jszx/uploadfile/200812/20081202024147201.pdf