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目的:试验研究了以二氧化硅粒子为载体固定辣根过氧化物酶的条件及固定化酶酶学特性。方法:通过溶胶-凝胶法制备了一种新的球形二氧化硅粒子并在其表面引入氨基,用戊二醛共价交联法固定化辣根过氧化物酶。分析了戊二醛的浓度、pH值、给酶量等因素对固定化酶活性的影响。结果:最佳固定条件是,戊二醛的溶度在3%(v/v),pH值在7.5,辣根过氧化物酶与二氧化硅微米颗粒的质量比为3.0∶50。固定化酶和游离酶反应的最佳pH值分别为7.5和7.0,最佳反应温度为50℃和40℃。结论:与游离酶相比,固定化酶具有优良的热稳定性和储存稳定性。
OBJECTIVE: To study the immobilization conditions of horseradish peroxidase (HRP) using silica particle as carrier and the enzymatic properties of immobilized enzyme. Methods: A new type of spherical silica particles was prepared by sol - gel method. Amino groups were introduced into the surface and horseradish peroxidase was immobilized by covalent cross - linking of glutaraldehyde. The effects of glutaraldehyde concentration, pH, amount of enzyme and other factors on immobilized enzyme activity were analyzed. Results: The best conditions for immobilization were glutaraldehyde with a solubility of 3% (v / v), a pH of 7.5 and a mass ratio of horseradish peroxidase to silica microparticles of 3.0: 50. The optimum pH of immobilized enzyme and free enzyme reaction were 7.5 and 7.0, respectively. The optimal reaction temperature was 50 ℃ and 40 ℃. CONCLUSION: Immobilized enzymes have excellent thermal and storage stability compared to free enzymes.