摘要
从海芋叶中分离和纯化过氧化物酶(POD),研究了不同参数超声波对其活性作用效果.结果表明,经不同频率、功率超声波处理不同时间后,酶活性均比未经超声波处理的活性低.将粗酶经硫酸铵分级沉淀、透析、DEAE-纤维素离子交换柱层析,在聚丙烯酰胺凝胶电泳图谱上显示出1条蛋白带.对纯酶进行动力学分析,结果表明,超声波处理后,POD的Km值变大,而vm ax值变化不大,说明超声波处理使酶与底物的亲和力降低.紫外吸收光谱、紫外差示光谱和荧光发射光谱的研究进一步表明,超声波处理使酶分子的构象发生变化,从而导致酶活性的改变.
In order to investigate the effects and action mechanism of ultrasound on activity of peroxidase（POD）,the enzyme was isolated from the leaves of Alocosia macrorrhiza and purified.The effects of ultrasound on the activity of the extracted enzyme were investigated with a variety of parameters of ultrasound.The results showed that when POD was disposed in ultrasound with different treatment,the activity of enzyme were lower than that of the enzyme not disposed in ultrasound.Further purified by ammonium sulfate fractionation,ion exchange chromatography on DEAE-cellulose,the single enzyme strap could be seen on the polyacrylamide gel electrophoresis diagram.The kinetics analysis showed that Km increased while vmax changed slightly after ultrasonic treatment.It indicated that the affinity of POD to its substrate decreased after being disposed in ultrasound.Analysis of ultraviolet absorption spectrum,ultraviolet differential spectrum and fluorescence emission spectrum of the enzyme indicated that the conformation of enzyme molecules had been altered after the treatment of ultrasound.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2007年第4期60-64,共5页
Journal of South China Agricultural University
基金
国家自然科学基金(10074016)
