Abstract: Oxidative refolding of denatured-reduced lysozyme at 5 mg•ml^-1 was performed by fed-batch operation with dilution additives(acetamide and glycerol).A kinetic model based on the three-state protein model was established to describe the refolding process, and the refolding and aggregation constants were obtained by fitting the model to the experimental data.At a lower GdmCl concentration (1 mol•L-1) and a proper acetamide concentration (5 mol•L^-1), the refolding of lysozyme could reach a yield higher than 80%.When GdmCl concentration was decreased, acetamide concentration should be properly increased to achieve a high refolding yield.This indicated the same effect of acetamide and GdmCl on protein refolding.In contrast, glycerol facilitated the refolding of lysozyme by enhancing the thermodynamic stability of native-state protein.This meant that glycerol was cooperative with GdmCl on facilitating protein refolding, and its addition could only give a high yield at a proper GdmCl concentration.

摘要： 研究了流加操作与稀释添加耦合辅助高浓度变性还原溶菌酶的复性，建立了相关的过程动力学模型.利用三态复性的过程模型对实验数据进行了拟合，获得了较好的拟合效果.利用模型分析了复性过程的动力学特性，结果表明，两者耦合可在较低的盐酸胍浓度(1 mol•L^-1)存在下使较高浓度的（5 mg•ml^-1）变性溶菌酶获得80%以上的复性收率；并且发现添加剂乙酰胺的辅助复性作用与盐酸胍相同，降低盐酸胍浓度，适当提高乙酰胺浓度即可使聚集体生成速率常数最小，酶的复性收率最大；但甘油与盐酸胍具有协同作用，只有在适当的盐酸胍浓度下添加甘油才可获得理想的复性效果.