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温度响应型HRP纳米胶囊用于酶的高效固定和回收

DOI: 10.3724/SP.J.1105.2013.12295, PP. 542-548

Keywords: 温度响应,纳米胶囊,N-异丙基丙烯酰胺,酶固定,酶回收

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Abstract:

以辣根过氧化物酶(horseradishperoxidase,HRP)为目标蛋白,N-异丙基丙烯酰胺(NIPAM)为单体,采用原位沉淀聚合法,在37℃,NIPAM和HRP质量比为6∶1的条件下,制备了粒径大小为13.7nm,zeta电位是(-3.7±0.3)mV的温度响应型辣根过氧化物酶纳米胶囊(nHRP).采用基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOFMS)、透射电镜(TEM)、动态光散射仪(DLS)和紫外-可见光分光光度计(UV-Vis)等对HRP的修饰度,nHRP的微观结构、粒径大小、催化活性、温度响应性能和热稳定性能进行了表征.研究表明,制备的辣根过氧化物酶纳米胶囊单分散性较好;在环境温度高于33℃时,nHRP出现响应变化,其粒径大小变化显著,且呈可逆性;nHRP相比HRP原酶热稳定性显著提高;50℃时,将nHRP高速离心,可使酶多次有效地分离和回收.

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