基于两亲性超分子的共价水凝胶辅助蛋白折叠

A novel monomeric species, 2-｛6-[3-(6-methyl-4-oxo-1, 4-dihydropyrimidin-2-yl)ureido]hexylcarbamoyloxy｝butyl acrylate(UPy-C₆-HBA), was synthesized while 4-hydroxybutyl acrylate(HBA)was introduced to the intermediate compound 2-(6-isocyanatohexyl ureido)-6-methyl-4[1H]pyrimidinone(UPy-C₆-NCO)which was obtained through the reaction of 2-aminomethyl-4-hydroxy-6-methylpyrimidine(MIS)with 1, 6-hexane diisocyanate (HDI). UPy-C₆-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-C₆-HBA monomer which in vitro refolding was facilitated via hydrophobic interactions. The effects of UPy-C₆-HBA concentration, hydrogel dosage, lysozyme concentration and refolding temperature on the refolding rate of lysozyme were meticulously investigated. Remarkably, the supramolecular hydrogel comprising UPy-C₆-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-C6-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.