玉米雄穗中多酚氧化酶的纯化与特性研究

Purification and characterization of polyphenol oxidase from corn tassel.

作者信息

Gul Guven R, Aslan N, Guven K, Matpan Bekler F, Acer O

机构信息

Science Teaching Section, Education Faculty, Dicle University, Diyarbakir, Turkey.

Molecular Biology and Genetics Departement, Science Faculty, Dicle University, Diyarbakir, Turkey.

出版信息

Cell Mol Biol (Noisy-le-grand). 2016 Nov 30;62(13):6-11. doi: 10.14715/cmb/2016.62.13.2.

DOI:10.14715/cmb/2016.62.13.2

PMID:28040055

Abstract

In this study, polyphenol oxidase (PPO) from corn tassel was extracted and partially purified through (NH4)2SO4 precipitation and gel filtration chromatography. Optimal temperatures for subsrates catechol and 4-methyl catechol were 40 °C and 30 °C, respectively. The optimal pH values were 8.0 for catechol and 6.0 for 4-methyl catechol. Catechol was the most suitible substrate (Km: 3.48 mM, Vmax: 1.0 Abs./ min.). The moleculer mass of PPO was determined as 158 kDa. In this work, sodium azide, ethylenediaminetetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) were found to inhibit the enzyme activity as 26.6 %, 22.2 % and 12.2 % ratio, respectively. Besides, the effects of carbohydrates such as sucrose, fructose, ribose and glucose on PPO activity were investigated. The enzyme was found to be activated 17 % by fructose and ribose, 16 % by glucose and 4 % by sucrose.

摘要

在本研究中，通过硫酸铵沉淀和凝胶过滤色谱法从玉米雄穗中提取并部分纯化了多酚氧化酶（PPO）。底物儿茶酚和4-甲基儿茶酚的最佳温度分别为40℃和30℃。儿茶酚的最佳pH值为8.0，4-甲基儿茶酚的最佳pH值为6.0。儿茶酚是最适合的底物（Km：3.48 mM，Vmax：1.0吸光度/分钟）。PPO的分子量测定为158 kDa。在本研究中，发现叠氮化钠、乙二胺四乙酸（EDTA）和十二烷基硫酸钠（SDS）分别以26.6%、22.2%和12.2%的比例抑制酶活性。此外，还研究了蔗糖、果糖、核糖和葡萄糖等碳水化合物对PPO活性的影响。发现该酶被果糖和核糖激活17%，被葡萄糖激活16%，被蔗糖激活4%。