摘要
采用聚丙烯酰胺凝胶迭氮法固定了辣根过氧化物酶 ,其作为催化剂用于荧光法测定过氧化物 ,并探讨了固定酶测定过氧化物的最佳条件 ,如溶液pH值、温度、反应时间、荧光剂用量等。结果表明 :酶固定化后 ,反应pH值范围变宽 ,为pH 5 .0～ 7.0和 7.5～ 9.0 ,最佳pH为 7.8左右 ;酶的热稳定性与储存稳定性也都得到提高 ,在室温下便可用固定酶进行长时间测定 ,且可较长时间保存。采用固定酶制成的酶柱用于HPLC测定过氧化物 ,固定酶可反复使用 ,简化了测定操作 。
Horseradish peroxidase ( HRP) was immobilized on polyacrylamide gel. The optimum pH, storage stability, thermal stability, optimum reaction time, and operational stability were studied for the immobilized HRP when used in the determination of peroxide. It is found that the effect of pH on immobilized enzyme is less sensitively than that on free enzyme. The optimum pH for free enzyme is about 5.8, however, immobilized enzyme has good action in a much more wide range of pH value with 5.0similar to7.0 and 7.5similar to9.0. It is also found that thermal and storage stability for immobilized is greater than free enzyme. Moreover, immobilized enzyme gel can be made into enzyme pillar used in high-performance liquid chromatographic to determine peroxides, and it has the,advantages of convenience and economy.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2003年第1期74-78,共5页
Chinese Journal of Analytical Chemistry
