| 丰水梨多酚氧化酶基因的克隆与原核表达 |
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| 引用本文： | 陈东生,王坤波,李勤,李娟,黄建安,刘仲华.丰水梨多酚氧化酶基因的克隆与原核表达[J].茶叶科学,2015(1):17-23. |
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| 作者姓名： | 陈东生 王坤波 李勤 李娟 黄建安 刘仲华 |
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| 作者单位： | 国家植物功能成分利用工程技术研究中心，湖南农业大学茶学教育部重点实验室，湖南 长沙 410128 |
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| 基金项目： | 国家自然科学基金，教育部“新世纪优秀人才支持计划” |
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| 摘 要： | 通过PCR方法克隆丰水梨PPO基因全长,并将其登录Genebank,登录号JQ861265。基因全长1782 bp,无内含子,编码的PPO属于亲水性蛋白质,无跨膜结构,含有593个氨基酸,分子量约为65.8 k Da,理论等电点为8.4。N端含有一段由47个氨基酸组成的转运肽。去除转运肽的成熟PPO分子量为60.8 k Da,理论等电点为6.69。PPO中含有两个铜离子结合区,主要位于PPO二级结构中的α-螺旋区域中。原核诱导目的蛋白在诱导3~6 h后积累量较大。诱导蛋白(PPO前体和成熟PPO)均能氧化儿茶素形成茶黄素。
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| 关 键 词： | 多酚氧化酶 基因克隆 原核表达 酶促合成 茶黄素 |
The Cloning and Prokaryotic Expression of Polyphenol Oxidase Gene in Pear (Pyrus Pyrifolia Nakai) |
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| CHEN Dongsheng,WANG Kunbo,LIQin
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LI Juan,HUANG Jian′an,LIU Zhonghua.The Cloning and Prokaryotic Expression of Polyphenol Oxidase Gene in Pear (Pyrus Pyrifolia Nakai)[J].Journal of Tea Science,2015(1):17-23. |
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| Authors: | CHEN Dongsheng WANG Kunbo LIQin LI Juan HUANG Jian′an LIU Zhonghua |
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| Abstract: | The polyphenol oxidase (PPO, GenBank accession No.JQ861265)geneswere clonedby PCRfromPyrus Pyrifolia Nakai.The full length of PPO gene was 1782bp without introns, coding a precursor peptide.PPO precursor consists of 593 amino acids with a molecular weight of about 65.8kDa. It has a theoretical PI of 8.4 and has a transit peptide consisting of 47 amino acids. The mature PPO without transit peptide consists of 547 amino acids with a molecular weight of about 60.8kDaand a theoretical PI of 6.69.There are 2 Cu-binding domain in theα-helix of PPOs. The protein accumulation got a peak in 3-6 hours.The induced proteins (precursor PPO and mature PPO)can oxidize catechins into theaflavinsin vitro. |
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| Keywords: | polyphenol oxidase gene clone prokaryotic expression enzymatic synthesis theaflavins |
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