Fabrication of a porous polymer membrane enzyme reactor and its enzymatic kinetics study in an artificial kidney model
Lili Liua,c, Juan Qiaoa,b, Hongyi Zhangc, LiQia,b
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, PR China.
A porous polymer membrane-based d-amino acid oxidase (DAAO) reactor was developed that mimicked enzymatic activity in a renal ischemia model. Using glycidyl methacrylate as a biocompatible reactive monomer, poly(styrene-glycidyl methacrylate) was synthesized via a reversible addition fragment chain transfer polymerization technique. The prepared porous polymer membrane was used as a support to effectively immobilize DAAO. Compared to DAAO modified on nonporous polymer membrane and free DAAO in solution, the constructed porous polymer membrane-based DAAO enzyme reactor displayed 3-fold and 19-fold increase in enzymolysis efficiency, respectively. In addition, a chiral ligand exchange capillary electrophoresis system for DAAO was used to study DAAO enzymatic kinetics with d,l-methionine as the substrate. The proposed porous polymer membrane-based enzyme reactor showed excellent performance both on reproducibility and stability. Moreover, the enzyme reactor was successfully applied to mimic DAAO activity in a renal ischemia model. These results demonstrated that the enzyme could be efficiently immobilized onto a porous polymer membrane as an enzyme reactor and has great potential in mimicking the enzymatic activity in kidney.
Keywords: D-amino acid Oxidase immobilization, Porous polymer membrane, Enzyme activity mimic, Artificial kidney.