固定相分子臂对溶质保留和蛋白质复性的影响以及离子交换色谱复性溶菌酶-分析化学专业论文.docxVIP

4小时。在此色谱条件下，还原变性溶菌酶的质量回收率和活性回收率均达到 4小时。在此色谱条件下，还原变性溶菌酶的质量回收率和活性回收率均达到 最高值。对比了两种不同的变性条件一透析和未透析法，对复性溶菌酶的影响， 透析比未透析法复性溶菌酶的质量回收率基本相同，透析比未透析法的活性回 收率高很多，这是由于透析法过程中除去了变性剂后，变性溶菌酶有一部分折 叠形成具有局部空间结构的折叠中间体，只是没有生物活性，在色谱复性时， 这些折叠中间体提高了溶菌酶J下确折叠的几率。 关键词：计量置换理论，间隔臂，DIOL疏水色谱，rhlNF吖，溶菌酶，芳香醇 同系物，复性，离子交换色谱OVCX) ABSTRACTStationary ABSTRACT Stationary and mobile phases in liquid chromatography(LC)is the most important factors effecting the solute retention．So the studies the effect of stationary phase on the retention of solute has great significance In Was used stoichiometric displacement theory for retention(SDT-R)to study the effect of stationary phase the solute retention，protein refolding and the refolding of the reduced／denatured lysozyme by both hydrophobic chromatography(HIC)and weak—cation exchange chromatography(WCX)． The thesis includes five parts aS the follows： 1．Review：Solute retetion mechanism in LC attracts many scientists attention that it is the foundation of separation and purification．It also provides directions for protein separation and purification，In this chapter solute retention mechanism in LC．protein refolding mechanism and new developments in protein refolding kinetics Were reviewed．It contains 61 references． 2．Stoichiometric displacement theory for retention，parameters in SDT-R，the phyical meanings of every paramters and the methods ofmeasuring four parameters in HIC were introduced． 3．Different ligands of chromatogrphic stationary phase Can effect solute retention． Four parameters log，、z、J and logcp ofproteins and aromatic alcohol homologues in SDT-R obtained from DIOL columns of HIC having same matrix and end group， but with and without molecular spacer were employed to find out the contribution of molecular spacer to protein and aromatic alcohol homologues retention．A conclusion was that the non-polar and polar amino acid residues of protein moleculars separately imeract with non-polar and polar groups of the ligands including molecular spacer,resulting in