固定化辣根过氧化物酶催化活性研究

Catalytic activity of immobilized horseradish peroxidase

目的:基于已有的研究成果,对固定化辣根过氧化物酶的催化活性,包括不同介质中固定化酶催化活性进行深入研究。同时研究了催化动力学参数Km。方法:通过固定化辣根过氧化物酶在水体系中的催化活性研究,分析了酶载量、各种常见金属离子、有机溶剂对HRP活性的影响,以及对酶促动力学参数进行了研究。结果:最佳催化条件下,酶载量%为91.7%,金属离子Cu2+、Ag+、Zn2+和有机溶剂乙醇、甲醇、丙酮对固定化辣根过氧化物酶催化活性显示出抑制作用。游离辣根过氧化物酶和固定化辣根过氧化物酶米氏常数Km值分别为2.89 mmol/L和3.65 mmol/L。结论:与游离酶相比,固定化酶具有优良的催化活性和对抑制剂良好忍耐性。

Objective：Based on existing research results,to further study the catalytic activity of the immobilized horseradish peroxidase,including the catalytic activity of the immobilized enzyme in different media.At the same time,catalytic kinetic parameters Km was studied.Methods： By studying on the catalytic activity of immobilized horseradish peroxidase in the water media,the enzyme-loaded amount was determined.The effects of common metal ions,organic solvents on the HRP activity were studied.The kinetics parameter Km was determined.Results： On optimal catalytic conditions,enzyme-loaded amount was 91.7%.Cu2＋,Ag＋ and Zn2＋ and ethanol,methanol,acetone showed inhibition effect on the catalytic activity of immobilized horseradish peroxidase.Michaelis constant Km values of free horseradish peroxidase and immobilized horseradish peroxidase were 2.89 mmol/L and 3.65 mmol/L,respectively.Conclusion： Compared with the free enzyme,immobilized enzyme has excellent catalytic activity and good tolerance to inhibitors.