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当前位置: 首页 > 文章 > 壳聚糖固定β-半乳糖苷酶及其应用稳定性研究 食品科学 2009,30 (1) 222-227
Position: Home > Articles > Immobilization ofβ-Galactosidase with Chitosan Microspheres and Its Operational Stability FOOD SCIENCE 2009,30 (1) 222-227

壳聚糖固定β-半乳糖苷酶及其应用稳定性研究

作  者：
范家佑;郁建平;余广进
单  位：
贵州大学生命科学学院
关键词：
壳聚糖微球;固定化;β-半乳糖苷酶;操作稳定性
摘  要：
以壳聚糖微球为载体,戊二醛为交联剂,固定β-半乳糖苷酶对β-半乳糖苷酶的固定化条件及固定化酶的各种性质进行研究,确定酶固定的最适条件为:用pH6.5的P-E-M缓冲液浸泡10h,25℃壳聚糖微球与0.5%戊二醛交联12h以上,4℃下酶与壳聚塘微球固定12h以上,酶活力回收率可达67%。固定化酶的最适温度为40℃左右,最适pH7.0。通过双倒数法求回归方程,求得酶动力学参数K_m值为0.613mmol/ml。固定化酶稳定性好,可以重复使用。将该固定化酶应用于乳糖分解实验和作为柱层析介质连续分解乳糖,分批反应6批次,乳糖水解率保持在90%以上,连续水解20d,乳糖水解率仍然可保持在75%以上。
译  名：
Immobilization ofβ-Galactosidase with Chitosan Microspheres and Its Operational Stability
作  者：
FAN Jia-you YU Jian-ping YU Guang-jin (College of Life Science,Guizhou University,Guiyang 550025,China)
关键词：
chitcsan;;microspheres;;immobilization;;β-galactosidase;;operational stability
摘  要：
β-Galactosidase was immobilized on chitosan microspheres with glutaraldehyde by cross-linking reaction,the immobilezation conditions and characterization of the immobilized enzyme were studide.The optimal conditions for immobi- lization were as follow:in pH 6.5 P-E-M solution for 10 h,chitosan micrnsphere was treated with 0.5% glutaraldehde at 25℃for 12h,the solution ofβ-galactosidase was immobilized on the carrier at 4℃for 12 h,enzyme activity recovery was 67%.Optimal temperature and pH for the immobilized enzyme were about 40℃and 7.0 respectively,apparent michealis constant of immobilizedβ-galactosidas(?) K_m=0.613 mmol/L.The stability of immobilizedβ-galactosidase was high after storage.It exhibited excellent operational stability.With 5% lactose as the substrate,the lactose hydrolysis was maintained at 90% after 6 cycles;the lactose hydrolysis was kept at 75% when the immobilized lactase was applied in the column chromatography reactor after 20 days.

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