乌贼墨中多酚氧化酶的分离及纯化

Purification of polyphenol oxidase from cuttlefish ink

用pH 7.2磷酸提取缓冲液从乌贼墨汁中提取多酚氧化酶。提取的粗酶液经 30 %～ 80 %饱和度硫酸铵分级沉淀、透析、DEAE SepharoseCL 6B柱层析和SephadexG 15 0柱层析后 ,得到纯化的多酚氧化酶 ,纯化倍数为 10 .78。纯化后的酶液经聚丙烯酰胺凝胶电泳 ,分别采用银染色和多酚氧化酶活性染色均显示一条电泳带。

Polyphenol oxidase (PPO) was extracted from the cuttlefish ink with pH 7.2 sodium phosphate extraction buffer. The crude enzyme extract was fractionated with solid ammonium sulfate of 30%-80% saturation. After dialysis, the enzyme solution was purified by DEAE Sepharose CL 6B column chromatography and Sephadex G 150 column chromatography respectively. A 10.78 fold purification of PPO was obtained. The result from polyacrylamide gel electrophoresis of purified enzyme indicated that only one band was found using silver stain and PPO activity stain, respectively.