β-环糊精促进变性-还原溶菌酶复性

REFOLDING OF DENATURED-REDUCED LYSOZYME FACILITATED BY β-CYCLODEXTRIN

研究了 β -环糊精 (β -CD)和表面活性剂十六烷基三甲基溴化铵 (CTAB)作为复性液添加剂对促进变性 -还原溶菌酶的氧化复性 .β -CD自身具有抑制肽链分子间聚集 ,提高复性率的作用 ,但其相对于变性酶的浓度需达到 30 0 0～ 5 0 0 0倍以上 ,否则作用效果不明显 .与其相比 ,利用CTAB和 β -CD联合作用系统具有更佳的促进复性效果 ,在适宜的盐酸胍浓度存在下可使较高浓度的变性 -还原酶实现完全复性 .确定了CTAB +β -CD系统促进变性酶复性的最佳操作

The oxidative refolding of denatured-reduced lysozyme in the presence of cetyl trimethyl ammonium bromide (CTAB) and/or β-cyclodextrin (β-CD) was investigated. The renaturation yield was improved in the presence of β-CD which inhibited the hydrophobic interaction between the unfolded protein molecules. However, β-CD concentration should be 3000-5000 times higher than the denatured enzyme to achieve a significant refolding effect. In contrast to β-CD, the refolding system with both CTAB and β-CD showed more favorable effect on the enzyme renaturation. With a properly enhanced guanidine hydrochloride concentration, the enzymatic activity of the denatured-reduced lysozyme could be completely regained. In addition, the optical pH (8.5) and temperature (37°C) was established for the CTAB+β-CD renaturation system.