利用温敏性凝胶的缓释作用促进变性溶菌酶复性

Enhanced Refolding of Denatured-Reductive Lysozyme Assisted by Control-Release of Temperature-Responsive Gel

为了开发温敏凝胶在蛋白质复性方面的方法,利用N-异丙基丙烯酰胺温敏凝胶具有的缓释作用,对变性蛋白质进行了复性.考察不同合成条件下凝胶的性能,结果表明,在制胶温度为22℃、凝胶单体浓度为1000/、交联度为500/制得的凝胶,具有较好的机械强度、较大的溶胀倍率、缩水倍率及良好的温敏性能.凝胶的缓释作用对变性还原溶菌酶辅助复性的影响表明,在复性酶终浓度为0.5 mg/mL时,凝胶辅助蛋白质的复性收率可达6800/,比直接稀释复性(收率为5700/)高1900/.特别是对高浓度蛋白质的复性(1.5 mg/mL),凝胶辅助复性也获得了很好的效果,复性收率高出直接稀释复性3700/,表现出温敏凝胶在实际蛋白质复性中的应用前景.

The control-release effect of ploy-N-isopropyl acrylamides （PNIPAAm） was applied to the refolding of denatured lysozyme. The physical properties of PNIPAAm were studied and the result indicates that the PNIPAAm prepared with monomer concentration of 10% and crosslinking density of 5% at 22℃ has good mechanical strength, satisfactory swelling and deswelling ratios, and good temperature-responsive performance. The refolding of denatured-reductive lysozyme assisted by this gel was investigated. The refolding yield reached 68%, which was 19% increase as compared with that of dilution refolding （57%）. Even in denatured protein so- lution of high concentration （ 1.5 mg/mL） , the control-release of the gel enhanced the refolding yield by 37%. Therefore, the control-release effect of the temperature-responsive gel can provide significant enhancement of the refolding yield to the denatured protein.