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Refolding of denatured carbonic anhydrase assisted by amide and refolding kinetics

LU Chenxing;XIAO Qingmin;DONG Xiaoyan

  

  • Online:2005-08-25 Published:2005-08-25

酰胺类物质辅助变性碳酸脱水酶复性及动力学

陆晨星;肖庆敏;董晓燕   

  1. 天津大学化工学院生物化工系,天津 300072

Abstract: The effect of various amides on the refolding of denatured carbonic anhydrase B (CAB) was investigated at low guanidine hydrochloride (GdmCl) concentration.The competitive model of first-order folding and third-order aggregation was found to well describe the dynamic refolding process in the presence of the additives. The results indicated that the denatured CAB could be properly refolded in the presence of the amides, and the effect of the amides on assisting CAB refolding was found kinetically similar to that of GdmCl. The refolding rate constant kN decreased with increasing additive concentration, while there was an optimum additive concentration where a minimum aggregation rate constant kA was obtained. At this additive concentration, the value of the ratio of kN to kA was maximum, leading to the maximal refolding yield of CAB.

摘要: 研究了酰胺类物质对促进变性牛碳酸脱水酶(CAB)的复性作用,考察了在较低盐酸胍浓度下添加剂浓度对复性收率的影响,并利用三级反应动力学方程对酰胺类化合物辅助CAB复性的动力学过程进行了深入分析.结果表明,含有酰胺基团的多种添加剂均能有效促进CAB复性;酰胺化合物与盐酸胍的作用相似,即复性速率常数kN值均大于无添加剂时的自发复性,且随添加剂浓度增加而降低;但聚集体生成速率常数kA值则在最佳添加剂浓度下出现最小值,从而使kN/kA值最大,复性收率最大.