阳离子交换纳晶胶分离乳过氧化物酶的研究  

Separation of lactoperoxidase using cation exchange nano-cryogels

作　　者：张颂红 赵欣怡 楼小玲 沈绍传 贠军贤 ZHANG Songhong;ZHAO Xinyi;LOU Xiaoling;SHEN Shaochuan;YUN Junxian(State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology,College of Chemical Engineering,Zhejiang University of Technology,Hangzhou 310032,Zhejiang,China)

机构地区：[1]浙江工业大学化学工程学院,绿色化学合成技术国家重点实验室培育基地,浙江杭州310032

出　　处：《化工学报》2024年第7期2574-2582,共9页CIESC Journal

基　　金：国家自然科学基金项目(22078296,21576240);浙江省自然科学基金重大项目(LD21B060001)。

摘　　要：乳过氧化物酶(lactoperoxidase,LP)因具有重要的生物学功能及广谱的抑菌活性而在食品、医药和化工等领域受到广泛关注。针对目前LP分离纯化方法中普遍存在的选择性差、成本高以及分离材料易污染等问题,以甲基丙烯酸羟乙酯(HEMA)及聚甲基丙烯酸缩水甘油酯纳凝胶(PGN)为复合基材,利用低温冷冻聚合和结晶致孔原理制备了pHEMA/PGN纳晶胶基质,然后接枝功能单体2-丙烯酰氨基-2-甲基-1-丙磺酸(AMPSA)得到具有阳离子交换功能的pHEMA/PGN纳晶胶介质,以用于从乳清中层析分离LP。考察了纳晶胶介质的基础性能,并重点探究了缓冲液pH对层析性能的影响。结果表明:制备所得纳晶胶介质具有分布均匀的超大孔结构、优良的渗透性能和传质性能,对模型蛋白溶菌酶的吸附容量为4.41 mg·ml^(-1);在最优分离条件下所得目标蛋白LP纯度约为96.0%,比活力为27.03 U·mg^(-1),酶活性回收率为86.5%,分离效果良好,表明该纳晶胶介质在层析分离方面具有较大的应用潜力。Lactoperoxidase(LP)has received widespread attention in the fields of food,medicine and chemical industry because of its important biological functions and broad-spectrum antibacterial activity.In view of the shortcomings of traditional LP separation and purification methods,such as poor selectivity,high cost and easy contamination of separated materials,2-hydroxyethyl methacrylate and poly(glycidyl methacrylate)nanogels(PGN)were used as composite substrates to prepare poly(2-hydroxyethyl methacrylate)composite cryogel embedded with PGN nanogels(pHEMA/PGN-based cryogel)by using the principle of low-temperature freezing polymerization.Then the pHEMA/PGN-based cryogel was grafted with the functional monomer 2-acrylamino-2-methyl-1-propanesulfonic acid to obtain pHEMA/PGN cation exchange nano-cryogel(pHEMA/PGN nano-cryogel)for the chromatography separation of LP from whey.The basic properties of pHEMA/PGN nano-cryogel were investigated,and the effect of buffer pH on chromatography performance was emphatically explored.The results showed that the obtained pHEMA/PGN nano-cryogel had large pore structure with uniform distribution,high permeability,excellent mass transfer performance,and strong adsorption capacity for lysozyme reaching 4.41 mg·ml^(-1).The pHEMA/PGN nano-cryogel was used to isolate LP in whey,the purity was about 96.0%,the specific enzyme activity was 27.03 U·mg-1,and enzyme activity recovery was 86.5%under the optimal conditions.The excellent separation performance indicated that the pHEMA/PGN nano-cryogel has good potential in the field of lactoperoxidase separation from whey.

关 键 词：乳过氧化物酶 晶胶介质 分离 离子交换 

分 类 号：TQ028.8[化学工程]