高性能胰蛋白酶分子表面印迹材料的制备及其大分子识别特性研究  

Preparing trypsin surface-imprinted material with high performance in aqueous medium and researching its recognition character

作　　者：陈涛[1] 门吉英[1] 高保娇[1] 

机构地区：[1]中北大学化学工程系,山西太原030051

出　　处：《化学研究与应用》2016年第5期636-643,共8页Chemical Research and Application

基　　金：山西省青年科学基金项目(2014021015-1)资助

摘　　要：以交联聚乙烯醇(CPVA)微球为基质,以甲基丙烯酸(MAA)为功能单体,在中性水介质中制备了高性能胰蛋白酶(TRY)大分子表面印迹材料。首先通过CPVA微球表面的羟基与甲基丙烯酰氯之间的酯化反应,将大量可聚合双键甲基丙烯酰基(MAO)引入微球表面,获得改性微球MAO-CPVA。在中性水溶液中,单体MAA的羧基几乎发生完全电离,凭借强烈的静电相互作用,MAA自动结合在碱性蛋白TRY大分子周围,形成单体-模板复合体。水溶液中过硫酸铵/亚硫酸氢钠引发体系所产生的自由剂,使包围在TRY周围的单体MAA与交联剂N,N'-亚甲基双丙烯酰胺(MBA)在微球MAO-CPVA表面发生接枝交联聚合,而模板TRY则被包裹在交联网络之中,除去模板后便得到了TRY表面印迹微球MIP-PMAA/CPVA。深入考察了该表面印迹微球的大分子识别性能。研究结果表明,表面印迹微球MIP-PMAA/CPVA对TRY具有优良的结合亲和性,结合容量高达84mg·g^(-1)(3.6μmol·g^(-1));对模板蛋白TRY具有特异的识别选择性,相对于另一种碱性蛋白木瓜蛋白酶,印迹微球对TRY的选择性系数高达21.62。In this study,a trypsin( TRY) surface imprinted material with high performance was prepared successfully in aqueous me-dium with crosslinked polyvinyl alcohol(CPVA)microspheres as matrix and with methacrylic acid(MAA)as functional monomer. Methylacryloyl group(MAO),in whose structure a polymerizable double bond was contained,was chemically introduced onto the surfaces of CPVA microspheres,resulting in the immobilized microspheres MAO-CPVA. In neutral aqueous solution,the carboxyl group of MAA was nearly completed dissociated so that the monomer molecules of MAA would be combined around the TRY macro-molecule as basic protein by right of strong electrostatic interaction and the complex of monomer MAA-template TRY would be formed. The initiating system of ammonium persulfate/sodium bisulfite produces free radicals,and these radicals initiated MAA a-round TRY and the crosslinker N,N′-methylene bisacrylamide( MBA) to produce graft-crosslinking polymerization. While macromol-ecules of TRY were wrapped within the crosslinked networks,obtaining TRY surface-imprinted material MIP-PMAA/CPVA after re-moving the template macromolecules. The macromolecular recognition property of the surface-imprinted microspheres MIP-PMAA/CPVA was investigated in depth. The experimental results showed that the imprinted microspheres MIP-PMAA/CPVA had excellent binding affinity for TRY, and the binding capacity reaches to 84mg·g-1 ( 3. 6μmol·g-1 ) . More importantly, microspheres MIP-PMAA/CPVA possessed specific recognition selectivity towards TRY,and in comparison with basic protein,papain,the coefficient of selectivity of imprinted microspheres for TRY actually were up to 21. 62.

关 键 词：胰蛋白酶 甲基丙烯酸 表面印迹 碱性蛋白质 大分子识别 

分 类 号：O631[理学—高分子化学]