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; College of Chemistry & Environmental Science, Hebei University, Baoding, 071002, PR China.
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; School of Chemical Sciences, University of Chinese Academy of Sciences, Beijing, 100049, PR China.
Talanta. 2020 Aug 15;216:120963. doi: 10.1016/j.talanta.2020.120963. Epub 2020 Mar 31.
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.
基于多孔聚合物膜的 D-氨基酸氧化酶（DAAO）反应器被开发出来，模拟了肾缺血模型中的酶活性。使用甲基丙烯酸缩水甘油酯作为生物相容性反应性单体，通过可逆加成-断裂链转移聚合技术合成了聚苯乙烯-甲基丙烯酸缩水甘油酯。制备的多孔聚合物膜用作有效固定 DAAO 的载体。与非多孔聚合物膜上修饰的 DAAO 和溶液中的游离 DAAO 相比，构建的基于多孔聚合物膜的 DAAO 酶反应器的酶解效率分别提高了 3 倍和 19 倍。此外，采用手性配体交换毛细管电泳系统，以 d,l-蛋氨酸为底物研究 DAAO 的酶动力学。所提出的基于多孔聚合物膜的酶反应器在重现性和稳定性方面均表现出优异的性能。此外，该酶反应器成功应用于模拟肾缺血模型中的 DAAO 活性。这些结果表明，酶可以有效地固定在多孔聚合物膜上作为酶反应器，在模拟肾脏中的酶活性方面具有很大的潜力。
