人工伴侣条件下谷胱甘肽浓度对溶菌酶复性的影响

Effect of Glutathion Concentration on Lysozyme Refolding Assisted by Artificial Chaperones

利用盐酸胍变性、二硫苏糖醇还原的溶菌酶为模型蛋白,考察人工伴侣一步法辅助溶菌酶复性时的最佳谷胱甘肽浓度。结果表明,当复性体系中溶菌酶浓度不同时,得到最大复性收率的GSSG与GSH的浓度有所差异。酶的质量浓度在0.06,0.1,0.2 mg/mL的条件下,随酶带入的还原型DTT的作用可以不予考虑,GSSG与GSH物质的量浓度比维持在1∶1时可以得到最大复性收率;酶的质量浓度在0.6 mg/mL的条件下,应当考虑DTT的作用,此时,需将变化后的GSSG与GSH物质的量浓度比维持在1∶1时,才能得到较大复性收率。

In order to explore the best concentration of glutathione in the test assisting with artificial chaperones by one-step method, the hydrochloric acid-Denatured and DTT-Reduced Lysozyme are taken as the model protein. The result indicates that the ratio of GSSG and GSH varies because of different concentrations of lysozyme in refolding buffer. Without considering the effect of reduced DTT, the best refolding yield could be obtained when the concentrations of lysozyme are 0.06, 0.1 and 0.2 mg/mL and the GSSG and GSH remains 1：1. When the lysozyme concentration is 0. 6 mg/mL, the effect of reduced DTT should be considered, the better refolding yield can be obtained while the transformed GSSG and GSH remains 1 ： 1.