食品科学 ›› 2007, Vol. 28 ›› Issue (12): 297-300.

• 生物工程 • 上一篇    下一篇

雪梨果肉多酚氧化酶的活性研究与纯化

 李桂琴, 刘坤, 张玉星   

  1. 河北经贸大学生物科学与工程学院; 河北农业大学园艺学院 河北石家庄050061河北农业大学园艺学院; 河北保定071001; 河北石家庄050061;
  • 出版日期:2007-12-15 发布日期:2011-11-22

Characteristic and Purification of Polyphenol Oxidase from Snow Pear Flesh

 LI  Gui-Qin, LIU  Kun, ZHANG  Yu-Xing   

  1. 1.Biological Science and Engineering Institute, Hebei University of Economics and Business, Shijiazhuang 050061, China; 2.College of Horticulture, Agricultural University of Hebei, Baoding 071001, China
  • Online:2007-12-15 Published:2011-11-22

摘要: 以雪梨果实为试材,通过以邻苯二酚为底物测定氧化产物的方法,测定温度、pH值、抑制剂、不同成熟期及不同部位对PPO活性的影响。结果表明:最适pH值为6.4;最适温度为25℃;不同成熟期PPO的活性不同;1mol/L的亚硫酸钠为较好的抑制剂;果肉内部PPO活性最高,果肉中部的PPO活性最低。粗提液又经30%硫酸铵沉淀去杂和80%饱和度硫酸铵析出、Sephadex-G200柱层析及HiPrepTM16/10QXL离子柱层析,得到电泳均一的PPO,分子量43kD。

关键词: 雪梨, 多酚氧化酶(PPO), 层析

Abstract: The polyphenol oxidase (PPO) in snow pear flesh was extracted and purified. The results showed that the optimum pH for PPO is 6.4; the optimum temperature for PPO is 25 ℃; the PPO activity changed at different stages; the 1 mol/L sodium sulfite is a better inhibitor and the PPO activity is largest in the interior of pear fleash.Oxidase was purified from snow pear flesh by four steps: Ammonium sulfate fractionation with saturation 30%, ammonium sulfate fractionation with saturation 80%, Sephadex-G200 filtration and HiPrepTM16/10 QXL column chromatography in turn. Through these steps, PPO was purified to electrophoretic homogeneity. SDS-PAGE showed the protein as a single polypeptide with a molecular weight of about 43.0 kD.

Key words: pear, phenol oxidize (PPO), chromatography