α-Glucosidase immobilization on functionalized Fe_3O_4 magnetic nanoparticles for screening of enzyme inhibitors  

作　　者：Guorong Cheng Junpeng Xing Zifeng Pi Shu Liu Zhiqiang Liu Fengrui Song 

机构地区：[1]National Center of Mass Spectrometry in Changchun &Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences [2]University of Science and Technology of China

出　　处：《Chinese Chemical Letters》2019年第3期656-659,共4页中国化学快报（英文版）

基　　金：funded by the National Natural Science Foundation of China(Nos.81473537,81773690,21673219)

摘　　要：Magnetic nanoparticles(MNPs) are widely used for the immobilization of enzyme owing to the unique properties such as good biocompatibility and rapid separation. Herein, we used Fe_3O_4 magnetic nanoparticles(Fe_3O_4 MNPs) as the carrier core with(3-aminopropyl)triethoxysilane(APTES)modification by our approach, in which a-glucosidase was stereoscopically immobilized on the surface of Fe_3O_4 MNPs via covalent binding. The result of immobilization was characterized by scanning electron microscope(SEM) and fourier transform-infrared spectroscopy(FT-IR). Then we optimized some key parameters of the immobilization reaction, including the ratio of MNPs to enzyme, GA concentration,crosslinking time and immobilization time. Moreover, under the optimal conditions, pH tolerance,thermo stability and reusability of the immobilized enzyme were investigated and compared with the free one. In order to evaluate the change of the affinity of the enzyme to its specific substrate after immobilization, the Michaelis-Menten constant(K_m) was also studied. Finally, the immobilized α-glucosidase combining with high performance liquid chromatography-tandem mass spectrometry technique(HPLC-MS/MS) was applied to screen and identify eight inhibitors from Polygonum cuspidatum extract. These results indicated that the established method had the broad prospects for biotechnological applications.Magnetic nanoparticles(MNPs) are widely used for the immobilization of enzyme owing to the unique properties such as good biocompatibility and rapid separation. Herein, we used Fe_3O_4 magnetic nanoparticles(Fe_3O_4 MNPs) as the carrier core with(3-aminopropyl)triethoxysilane(APTES)modification by our approach, in which a-glucosidase was stereoscopically immobilized on the surface of Fe_3O_4 MNPs via covalent binding. The result of immobilization was characterized by scanning electron microscope(SEM) and fourier transform-infrared spectroscopy(FT-IR). Then we optimized some key parameters of the immobilization reaction, including the ratio of MNPs to enzyme, GA concentration,crosslinking time and immobilization time. Moreover, under the optimal conditions, pH tolerance,thermo stability and reusability of the immobilized enzyme were investigated and compared with the free one. In order to evaluate the change of the affinity of the enzyme to its specific substrate after immobilization, the Michaelis-Menten constant(K_m) was also studied. Finally, the immobilized α-glucosidase combining with high performance liquid chromatography-tandem mass spectrometry technique(HPLC-MS/MS) was applied to screen and identify eight inhibitors from Polygonum cuspidatum extract. These results indicated that the established method had the broad prospects for biotechnological applications.

关 键 词：Magnetic nanoparticles(MNPs) Α-GLUCOSIDASE IMMOBILIZATION HPLC-MS/MS Inhibitor SCREENING POLYGONUM cuspidatum 

分 类 号：TQ460.1[化学工程—制药化工]