苹果多酚氧化酶的纯化与表征(英文)

被引:3
作者
肖厚荣 [1 ,2 ]
杨红 [1 ]
蔡敬民 [1 ,2 ]
王中风 [1 ]
刘斌 [1 ,2 ]
张洁 [1 ]
吴克 [1 ,2 ]
机构
[1] 合肥学院生物与环境工程系
[2] 合肥学院应用酶学与酶工程重点实验室
关键词
苹果; 多酚氧化酶; 纯化; 表征;
D O I
暂无
中图分类号
S661.1 [苹果];
学科分类号
090201 ;
摘要
运用丙酮浸漬干燥、磷酸盐缓冲液提取、低温离心、硫酸铵沉淀、DEAE-Sephadex(A-50)、Sephadex(G-75)和DEAE-celluse(DE-52)层析等方法从苹果中分离获得一种新的含铜酶蛋白,该酶被命名为多酚氧化酶Ⅱ(polyphenoloxidaseⅡ,PPOⅡ),纯化倍数是215,纯化收率是23%。PAGE、SDS-PAGE和MALDI-TOF等技术用于测定所获的酶的纯度和分子量。在PAGE和SDS-PAGE均显示一条带,表明PPOⅡ只由一个亚基组成,且已达到单一组分(MALDI-TOF的结果更证实了这一点)。SDS-PAGE和MALDI-TOF的结果都表明PPO的分子量为38204Da。pH值对酶活性和稳定性研究的结果显示,从pH值4.0~7.0随着pH值的增加,酶活性也不断增加;从pH值7.0~11.0,酶活性不断降低。PPOⅡ的最适pH值为6.6最适温度为30℃。
引用
收藏
页码:41 / 44+25 +25
页数:5
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