槟榔果仁多酚氧化酶分离纯化及其酶特性研究  

Isolation, purification and characterization of polyphenol oxidase from areca nut kernel

作　　者：郭宇婷 潘永贵[1] 张正科[1] 张伟敏[1] GUO Yu-ting;PAN Yong-gui;ZHANG Zheng-ke;ZHANG Wei-min(College of Food Science and Technology,Hainan Universityl Haikou570228)

机构地区：[1]海南大学食品学院,海口570228

出　　处：《食品科技》2018年第11期235-239,245,共6页Food Science and Technology

基　　金：海南省自然科学基金项目(318MS014)

摘　　要：以槟榔果仁为试材,经pH6.8磷酸缓冲溶液(PBS)提取多酚氧化酶(PPO)粗酶液,采用(NH4)2SO4分级沉淀后,进一步通过DEAE-Sepharose Fast Flow和Phenyl Sepharose 6 Fast Flow柱层析,获得PPO纯化酶液。经鉴定其纯度达到电泳纯,相对分子质量为29.2 ku。在此基础上,对提取的PPO进行酶学特性研究,结果表明,槟榔果仁中PPO最适温度为20℃,最适pH为7.0;以邻苯二酚为底物,该酶的最大反应速度(Vmax)为140.84 U/mL·min,Km为3.22 mmol/L。Using areca nut kernel as test material, polyphenol oxidase(PPO) crude enzyme solution was extracted by pH 6.8 phosphate buffer solution(PBS). After(NH4)2 SO4 fractional precipitation, and then through DEAE-Sepharose Fast Flow and Phenyl Sepharose 6 Fast Flow column chromatography, the purified PPO solution was obtained, indicating the presence of PPO enzyme in the areca nut kernel. Its purity was confirmed to be pure by electrophoresis and its relative molecular mass was determined to be 29.2 ku. On this basis, the enzymatic characterization of the extracted PPO were studied, the results showed that the optimum temperature of PPO in areca nut kernel is 20 ℃, the optimum pH is 7.0, and the maximum reaction rate of the enzyme(Vmax) is 140.84 U/mL·min with catechol as the substrate, Km is 3.22 mmol/L.

关 键 词：槟榔果仁 PPO 分离纯化 酶特性 

分 类 号：R284.2[医药卫生—中药学]