超声波对海芋过氧化物酶催化活性影晌的机理.pdfVIP

第28 卷第4 期 华南农业大学学报 Vol. 28 , No.4 2∞7 年 10 月 loumal of South China Agricultural University Oct. 2007 超声波对海芋过氧化物酶催化活性影晌的机理 许可，黄卓烈，邱桂英，巫光宏，詹福建，何平 (华南农业大学生命科学学院，广东广州 510642) 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-将粗酶经硫酸续分级沉淀、适析、DEAE­ 纤维素离子交换柱层析，在聚丙烯-m胶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 Kιm 值变大，而 V怡1，阳川，m m 外差示光语和荧光发射光i谱昔的研究进一步表明，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- 关键词:超声波;过氧化物酶(POD) ;动力学;光谱 中固分类号:Q55 文献标i只码 :A 文章编号:1∞1-41l X( 2007) 04-0060-05 Effect of Ultrasound on the Activity of Peroxidase from Alocosia macrorrhiza Leaves and the Mechanism XU Ke , HUANG Zhuo-lie , QIU Gui-ying , WU Guang-hong , ZHAN Fu-jian , HE Ping 、 (College of Life Science , South China Agric. Univ. , Guangzhou 510642 , China) Abstract: 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 macro厅hizαand purified. The effects of ul- trasound on the activity of the extracted enzyme were investigated with a variety of parameters of ultra- sound. The results showed that when POD was disposed in ultrasound with different treatment , the activi- ty of enzyme were lower than that of the enzyme not disposed in ultrasound. Further purified by ammoni- um sulfate fractionation , ion exchange chromatography on DEAE-cellulose , the single enzyme strap could be seen on 出e polyacrylamide gel electrophoresis diagram. The kinetics analysis showed that Km in- creased while VmaX changed slightly after ultras