人工伴侣与凝胶过滤色谱耦合作用促进高浓度溶菌酶复性研究

Renaturation of high concentration lysozyme using gel filtration chromatography assisted by artificial chaperones

研究了人工伴侣系统,即十六烷基三甲基溴化铵(CTAB)和β 环糊精(β CD)促进高浓度变性 还原溶菌酶的氧化复性。确定CTAB与β CD的浓度比为1∶4,CTAB与变性溶菌酶的结合时间在10—50min内复性收率较高。同时发现,利用人工伴侣和凝胶过滤色谱的耦合作用可有效提高溶菌酶的复性收率,在连续复性操作条件下可使进料质量浓度为1mg/mL的变性酶获得87%的总复性收率。

Oxidative renaturation of denatured-reduced lysozyme assisted by the artificial chaperones, that was, cetyltrimethyl ammonium bromide (CTAB) and β-cyclodextrin (β-CD), was investigated. Experimental results indicate that the denatured enzyme is most favorably renatured at a molar ratio 1∶4 of CTAB to β-CD and a binding time of denatured lysozyme and CTAB between 10 and 50 min. Batch renaturation verified that the cooperation of the artificial chaperones and gel filtration chromatography significantly improved the recovery of lysozyme renaturation. Thus, a continuous renaturation approach assisted by the artificial chaperones in gel filtration chromatography was developed. The new method yielded 87% recovery of 1mg/mL lysozyme at continuous feeding.