中华寿桃果肉多酚氧化酶的纯化研究

Purification of Polyphenol Oxidase of ‘Zhonghuashoutao Peach Flesh

研究了影响中华寿桃果肉褐变的关键酶——多酚氧化酶(PPO)的纯化。结果表明,采用30%硫酸铵盐析、Phenyl Toyopearl柱层析、90%硫酸铵再次盐析、Sephadex G-25、DEAE-Sepharose柱层析等5步分离,可得到电泳均一的PPO,其纯化倍数为110倍,产率为0.36%,分子量约为66.0kD。

The polyphenol oxidase (PPO) in peach flesh was extracted and purified. The results showed that polyphenoloxidase was purified from Zhonghuashoutao peach flesh by five steps: Ammonium sulfate fractionation with saturation30%, Phenyl Toyopearl, ammonium sulfate fractionation with saturation 90%, Sephadex G-25 filtration, and DEAE-sepharoseanion column chromatography in turn. Through these steps, PPO was purified to electrophoretic homogeneity. SDS-PAGEshowed the protein as a single polypeptide with a molecular weight of about 66.0kD. The final purification fold and ratio ofrecovering protein are 110 and 0.36%, respectively.