基于磁性纳米粒子的两种蛋白酶固定化策略对蛋白水解效率的对比研究  

Comparative study on two immobilization strategies of protease based on magnetic nanoparticles and their proteolytic efficiency

作　　者：王晓萱 姚梦迪 杨适春 邵晨 杜超 赵华伟[1,2] WANG Xiaoxuan;YAO Mengdi;YANG Shichun;SHAO Chen;DU Chao;ZHAO Huawei(School of Food Engineering,Ludong University,Yantai 264025;Institute of Biological Nanotechnology,Ludong University,Yantai 264025)

机构地区：[1]鲁东大学食品工程学院,烟台264025 [2]鲁东大学生物纳米技术研究院,烟台264025

出　　处：《中国食品添加剂》2025年第3期92-100,共9页China Food Additives

摘　　要：为了比较木瓜蛋白酶的不同固定化方法,采用溶剂热法制备得到磁性纳米粒子,并通过硅烷化学进行表面氨基化,再分别通过聚丙烯酸原位聚合和丁二酸酐修饰两种方法实现纳米表面羧基化,进而通过酰胺化实现木瓜蛋白酶的固定化。对木瓜蛋白酶的两种固定化条件进行优化,比较两种木瓜蛋白酶的酶学性质,并对两种固定化酶的蛋白水解效果进行评价。结果表明经丁二酸酐修饰法固定化酶的最适pH为11.0,经聚丙烯酸原位聚合法固定化酶的最适pH为6.0,两者最适温度均为50℃。丁二酸酐修饰后固定化酶对酪蛋白的水解度可达40%左右,另一种仅25%左右。因此,丁二酸酐修饰后的磁性纳米粒子固定化酶的方法在酶学性质方面较优,对酪蛋白的水解度更高。In order to compare papain's two immobilization strategies,magnetic nanoparticles were prepared using a solvothermal method,followed by surface amination through silanization.Subsequently,carboxylation of the nanoparticle surface was achieved using two methods:in situ polymerization of polyacrylic acid and modification with succinic anhydride.This was followed by amidation for the immobilization of papain.The immobilization conditions of papain were optimized,and the enzymatic properties of the two types of immobilized papain were compared.The hydrolysis effects of the two immobilized enzymes were evaluated.The results indicated that the optimal pH for the enzyme immobilized via succinic anhydride modification was found to be 11.0,while the optimal pH for the enzyme immobilized via polyacrylic acid in-situ polymerization was 6.0,with the optimal temperature for both was 50℃.The degree of protein hydrolysis for the enzyme immobilized via succinic anhydride modification reached approximately 40%,whereas the other method achieved only around 25%.Therefore,the method utilizing succinic anhydridemodified magnetic nanoparticles for enzyme immobilization exhibited superior enzymatic properties and resulted in more thorough hydrolysis of casein.

关 键 词：固定化酶 磁性纳米粒子 酶学性质 水解度 

分 类 号：TS202.3[轻工技术与工程—食品科学]