磁性纳米氮化铝颗粒固定化β-葡萄糖苷酶的性质  

Properties of β-Glucosidase Immobilized on Magnetic Nanoparticles of Aluminum Nitride

作　　者：潘利华[1] 罗建平[1] 王贵娟[1] 徐学玲[1] 宛雯[1] 

机构地区：[1]合肥工业大学生物与食品工程学院生物工程系,安徽合肥230009

出　　处：《催化学报》2008年第10期1021-1026,共6页

基　　金：安徽省自然科学基金(050410301);安徽省青年教师资助计划(2006jp1022)

摘　　要：以戊二醛为交联剂,研究了磁性纳米氮化铝颗粒固定化β-葡萄糖苷酶的条件及固定化酶的结构特征,并考察了固定化酶的某些酶学性质.结果表明,在4.5ml磁性纳米氮化铝颗粒悬液(100mg/ml)中加入0.5ml戊二醛溶液(2%)超声波分散后,加入5mlβ-葡萄糖苷酶溶液(50mg/ml),于20℃,pH5.0和100r/min条件下固载3.5h,酶蛋白和酶活回收率分别为82.6%和78.4%.固定化β-葡萄糖苷酶的结构松散,不改变酶的结构特征.与游离酶相比,固定化酶对对硝基苯基-β-D-葡萄糖苷水解反应的最佳反应温度有所降低,最佳反应pH值有所升高,而米氏常数Km值有所增大,且具有良好的贮存稳定性和操作稳定性,表明磁性纳米氮化铝颗粒适合作为β-葡萄糖苷酶的固定化载体.The conditions for β-glucosidase immobilization with magnetic nanoparticles of aluminum nitride were optimized and the properties of immobilized β-glucosidase were investigated. The maximum immobilized protein recovery （82.6 % ） and activity recovery （78.4%） were obtained with β-glucosidase solution （5 ml, 50 mg/ml） and glutaraldehyde solution （0.5 ml, 2% ） added to sus- pension of magnetic aluminum nitride nanoparticles （4.5 ml, 100 mg/ml） and immobilized for 3.5 h at the temperature of 20 ℃ and pH value of 5.0. The structure of immobilized enzyme was incompact and not changed evidently. The Michaelis constant （Kin） for hydrolysis of 4-nitrophenyl-β-D-glucoside by the immobilized β-glucosidase, with good storage stability and operation stability, was 0.88 mmol/L at the optimal temperature of 55℃ and the optimal pH value of 5.0. The results suggested that the magnetic aluminum nitride nanoparticle is a good support for the immobilization of β-glucosidase.

关 键 词：磁性纳米颗粒 氮化铝 葡萄糖苷酶 固定化 戊二醛 

分 类 号：TB383.1[一般工业技术—材料科学与工程]