基于两亲性超分子的共价水凝胶辅助蛋白折叠  

Protein Folding Assisted by Covalent Hydrogel Based onAmphiphilic Supramolecular

作　　者：王嘉鑫 刘佳 秦金梅 NYV Mondele Mbola 孟子晖 薛敏 WANG Jiaxin;LIU Jia;QIN Jinmei;NYV Mondele Mbola;MENG Zihui;XUE Min(School of Chemistry and Chemical Engineering,Beijing Institute of Technology,Beijing 100190,China)

机构地区：[1]北京理工大学化学与化工学院,北京100190

出　　处：《高等学校化学学报》2023年第11期173-181,共9页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:21874009)资助。

摘　　要：采用2-氨基-4-羟基-6甲基嘧啶(MIS)和1,6-己二异氰酸酯(HDI)合成了中间体2-(6-异氰酸酯己基脲)-6-甲基-4[1H]嘧啶酮(UPy-C_(6)-NCO),并引入4-羟基丁基丙烯酸酯(HBA),合成了一种新型单体2-{6-[3-(6-甲基-4-氧代-1,4-二氢嘧啶-2-基)脲基]己烷氨甲酰氧基}丙烯酸丁酯(UPy-C_(6)-HBA).通过紫外光引发自由基聚合,使UPy-C_(6)-HBA和丙烯酰胺(AM)、N,N'-亚甲基双丙烯酰胺(Bis)进行共聚,制备了具有均匀多孔结构和高度溶胀性的物理化学双交联水凝胶.将溶菌酶包埋在含UPy-C_(6)-HBA单体的水凝胶基质中,通过疏水作用辅助溶菌酶进行体外折叠复性研究.结果表明,UPy-C_(6)-HBA含量、凝胶用量、溶菌酶浓度和复性温度对复性效果均有影响.当环境温度为25℃时,含UPy-C_(6)-HBA单体15%(质量分数)的水凝胶可以使0.25 mg/m L溶菌酶的复性率提高41%.研究结果表明,向凝胶体系中加入UPy-C_(6)-HBA单体显著提高了溶菌酶的复性率,对于高浓度变性溶菌酶的复性具有重要意义.A novel monomeric species,2-{6-[3-(6-methyl-4-oxo-1,4-dihydropyrimidin-2-yl)ureido]hexylcarbamo-yloxy}butyl acrylate(UPy-C_(6)-HBA),was synthesized while 4-hydroxybutyl acrylate(HBA)was introduced to the intermediate compound 2-(6-isocyanatohexyl ureido)-6-methyl-4[1H]pyrimidinone(UPy-C_(6)-NCO)which was ob-tained through the reaction of 2-aminomethyl-4-hydroxy-6-methylpyrimidine(MIS)with 1,6-hexane diisocyanate(HDI).UPy-C_(6)-HBA together with acrylamide(AM)and N,N'-methylenediacrylamide(Bis)was polymerized by UV-induced free radical polymerization method,yielding physically crosslinked hydrogels that possessed a homogenous porous architecture and pronounced swelling capacity.Lysozyme was incorporated within the hydrogel matrix containing UPy-C_(6)-HBA monomer which in vitro refolding was facilitated via hydrophobic interactions.The effects of UPy-C_(6)-HBA concentration,hydrogel dosage,lysozyme concentration and refolding temperature on the refolding rate of lysozyme were meticulously investigated.Remarkably,the supramolecular hydrogel comprising UPy-C_(6)-HBA at a mass fraction of 15%exhibited a remarkable 41%enhancement in the refolding rate of lysozyme at 25℃compared with the control.These findings unequivocally demonstrate the addition of UPy-C_(6)-HBA monomer to the hydrogel matrix significantly improved the refolding rate of lysozyme,which is of great significance for the refolding of high concentration denatured lysozyme.

关 键 词：2-{6-[3-(6-甲基-4-氧代-1 4-二氢嘧啶-2-基)脲基]己烷氨甲酰氧基}丙烯酸丁酯 超分子水凝胶 蛋白质折叠 溶菌酶 

分 类 号：O633[理学—高分子化学]