Towards cost-effective drug discovery: Reusable immobilized enzymes for neurological disease research.

Enzyme handling and utilization bears many challenges such as their limited stability, intolerance of organic solvents, high cost, or inability to reuse. Most of these limitations can be overcome by enzyme immobilization on the surface of solid support. In this work, the recombinant form of human cholinesterases and monoamine oxidases as important drug targets for neurological diseases were immobilized on the surface of magnetic non-porous microparticles by a non-covalent bond utilizing the interaction between a His-tag terminus on the recombinant enzymes and cobalt (Co2+) ions immobilized on the magnetic microparticles. This type of binding led to targeted enzyme orientation, which completely preserved the catalytic activity and allowed high reproducibility of immobilization. In comparison with free enzymes, the immobilized enzymes showed exceptional stability in time and the possibility of repeated use. Relevant K m , V max , and IC 50 values using known inhibitors were obtained using particular immobilized enzymes. Such immobilized enzymes on magnetic particles could serve as an excellent tool for a sustainable approach in the early stage of drug discovery. [Display omitted] • His-tagged ChEs & MAOs enzymes immobilization on magnetic particles via IMAC. • Enzymes retained catalytic properties post-immobilization. • Enhanced stability, enabling enzyme reusability. • Immobilized ChEs & MAOs application for facile inhibition assays. • Sustainable drug discovery with magnetic particle-immobilized enzymes. [ABSTRACT FROM AUTHOR]