Study on preparation of immobilized acetylcholinesterase microreactor and its application in CMM inhibitors screening / 中草药

ABSTRACT

Objective To prepare an immobilized acetylcholinesterase (AChE) microreactor and establish a rapid screening method for Chinese materia medica (CMM) acetylcholinesterase inhibitors (AChEIs). Methods A novel immobilized AchE microreactor was prepared by crosslinking with glutaraldehyde, using aminated magnetic microspheres as carrier. The characterizations were conducted by physicochemical properties and chromatographic performance. The immobilized AChE reactor was used to screen AChEIs from the Huperzia Serratum extracts. Results In the enzyme reaction system, the optimum substrate concentration was 50 μmol/L, and the incubation time was 5 min, respectively. IR characterization, specificity verification, enzyme kinetics, and stability study results all demonstrated the effectiveness of the enzyme reactor. The CMM AChEI, huperzine A, was obtained from the screening of H. Serratum extracts. Conclusion A high throughput screening method for AChEIs is established in this paper, which will be further applied and popularized.