乌贼墨中多酚氧化酶的分离及纯化

Purification of polyphenol oxidase from cuttlefish ink

摘要:

用pH7.2磷酸提取缓冲液从乌贼墨汁澡提取多酚氧化酶，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-150柱层析后，得到纯化的多酚氧化酶，纯化们数为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。

Abstract:

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.