块状凝胶的制备及其协助溶菌酶复性的研究  

Synthesis of hydrogel disk and its application in lysozyme refolding

作　　者：崔志芳[1] 关怡新[1] 姚善泾[1] 

机构地区：[1]浙江大学化学工程与生物工程学系,浙江杭州310027

出　　处：《浙江大学学报（工学版）》2004年第9期1212-1216,共5页Journal of Zhejiang University：Engineering Science

基　　金：国家自然科学基金资助项目(20076042);浙江省自然科学基金资助项目(201099).

摘　　要：利用水溶液聚合法合成温敏型聚N异丙基丙烯酰胺(PNIPA)块状凝胶,对其性能进行了表征并将其应用到溶菌酶的复性研究中.制得的PNIPA凝胶的低临界溶解温度为33℃,最大溶胀倍率为27,凝胶达到溶胀及缩水平衡需24h.加入270mg/mL凝胶时,溶菌酶复性后的比活力达到5450.6U/mg,与稀释复性相比提高了61.5%.对复性机理的分析表明:聚合物长链上的疏水基团与折叠中间体间的疏水相互作用是PNIPA凝胶协助复性的关键.A novel protein renaturation system using temperature sensitive hydrogel - poly (N-isopropylac-rylamide) (PNIPA) was developed. A lump of PNIPA gel was synthesized by solution polymerization. The low critical solution temperature of PNIPA was determined to be 33°C. The time for swelling equilibrium was about 24 h with swelling ratio being approximately 27. On adding of about 270 mg/mL gel disks into the refolding buffer, the specific activity of the denatured lysozyme in the buffer was 5450.6 U/mg, which increased about 61.5% compared with refolding by direct dilution. The refolding mechanism is further shown to be due to the inhibition of the hydrophobic interaction between the protein and the intermediates by the hydrophobic groups of the gel.

关 键 词：块状凝胶 制备 溶菌酶 复性 聚N-异丙基丙烯酰胺 温敏型凝胶 蛋白质 

分 类 号：TQ932[化学工程]