磁性交联β-葡萄糖苷酶聚集体的制备及性质  

The Preparation and Properties of Magnetic Cross-Linked β-Glucosidase Aggregates

作　　者：吴星涛[1] 薛原楷[1] 石贤爱[1,2] 

机构地区：[1]福州大学生物科学与工程学院,福州350108 [2]福建省医疗器械和医药技术重点实验室,福州350002

出　　处：《生物物理学报》2013年第10期769-778,共10页Acta Biophysica Sinica

基　　金：国家海洋公益性行业科研专项子课题(201205022-3);福建省发改委资助项目([2008]256)~~

摘　　要：采用磁性固定化酶技术制备磁性交联β-葡萄糖苷酶聚集体,优化了制备条件,并对其酶学性质进行了研究。正交试验结果表明,最优制备条件为:β-葡萄糖苷酶2 mg、沉降剂体积1 ml、磁性铁氧化物颗粒质量6 mg、戊二醛浓度15 mmol/L、交联时间2 h。在该条件下制备的磁性交联β-葡萄糖苷酶聚集体比活力可达6.44 U/mg,酶活回收率达80.5%。电镜观察显示磁性交联β-葡萄糖苷酶聚集体呈球状颗粒,单分散性好,直径约为50μm。与游离酶相比,磁性交联β-葡萄糖苷酶聚集体热稳定性及酸碱稳定性均有所提高,K m增大,操作稳定性良好,对磁场有较强的耐受性。The magnetic cross-linked 13-glucosidase aggregates （M-CLGAs） were prepared through the optimized programs and conditions. Furthermore, the enzymatic properties of M-CLGAs were also studied. From the results of orthogonal experiments, the optimal conditions would be 13-glucosidase mass 2 mg, sedimentation agent 1 ml, magnetic iron oxide particles 6 mg, glutaraldehyde 15 mmol/L, crosslinking time 2 h. Under the preparation condition, the specific activity of the prepared M-CLGAs could reach 6.44 U/mg, and the maximal recovery activity of M-CLGAs was 80.5%. It was observed through the SEM that the immobilized enzymes were spherical particles with good monodispersity, and the diameters of the particles were about 50 μm. Compared with the free enzyme, the M-CLGAs showed improved thermal stability and pH stability. Furthermore, M-CLGAs also exhibited good operational stability and magnetic stability, but the Kr, value was higher than that of the free enzyme.

关 键 词：磁性交联酶聚集体 Β-葡萄糖苷酶 固定化 酶学性质 稳定性 

分 类 号：Q814[生物学—生物工程]