title: Construction of a D-amino acid oxidase reactor based on magnetic nanoparticles modified by a reactive polymer and its application in screening enzyme inhibitors.
abstract: Developing facile and high-throughput methods for exploring pharmacological inhibitors of D-amino acid oxidase (DAAO) has triggered increasing interest. In this work, DAAO was immobilized on the magnetic nanoparticles, which were modified by a biocompatible reactive polymer, poly(glycidyl methacrylate) (PGMA) via an atom transfer radical polymerization technique. Interestingly, the enzyme immobilization process was greatly promoted with the assistance of a lithium perchlorate catalyst. Meanwhile, a new amino acid ionic liquid (AAIL) was successfully synthesized and employed as the efficient chiral ligand in a chiral ligand exchange capillary electrophoresis (CLE-CE) system for chiral separation of amino acids (AAs) and quantitation of methionine, which was selected as the substrate of DAAO. Then, the apparent Michaelis-Menten constants in the enzyme system were determined with the proposed CLE-CE method. The prepared DAAO-PGMA-Fe3O4 nanoparticles exhibited excellent reusability and good stability. Moreover, the enzyme reactor was successfully applied in screening DAAO inhibitors. These results demonstrated that the enzyme could be efficiently immobilized on the polymer-grafted magnetic nanoparticles and that the obtained enzyme reactor has great potential in screening enzyme inhibitors, further offering new insight into monitoring the relevant diseases.
authors: Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, P.R. China.
authors: Xiaoyu
authors: X
authors: Mu
authors: Juan
authors: J
authors: Qiao
authors: Li
authors: L
authors: Qi
authors: Ying
authors: Y
authors: Liu
authors: Huimin
authors: H
authors: Ma
mesh_terms: D000818:Animals; D019149:Bioreactors; D002850:Chromatography, Gel; D003605:D-Amino-Acid Oxidase / Q000037:antagonists & inhibitors*; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000032:analysis / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors*; D020128:Inhibitory Concentration 50; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D011109:Polymethacrylic Acids / Q000737:chemistry*; D059027:Recycling; D017550:Spectroscopy, Fourier Transform Infrared; D034421:Sus scrofa; D013997:Time Factors
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D011109:Polymethacrylic Acids; C042535:polyglycidyl methacrylate; D003605:D-Amino-Acid Oxidase