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从朝鲜蓟中分离多酚氧化酶 (PPO) cDNA 并分析其在受伤朝鲜蓟头部的表达。

Isolation of a polyphenol oxidase (PPO) cDNA from artichoke and expression analysis in wounded artichoke heads.

机构信息

Istituto di Scienze delle Produzioni Alimentari-CNR, Unità di Lecce, Via Prov.le Lecce-Monteroni, 73100 Lecce, Italy.

出版信息

Plant Physiol Biochem. 2013 Jul;68:52-60. doi: 10.1016/j.plaphy.2013.03.020. Epub 2013 Apr 11.

Abstract

The polyphenol oxidase (PPO) enzyme, which can catalyze the oxidation of phenolics to quinones, has been reported to be involved in undesirable browning in many plant foods. This phenomenon is particularly severe in artichoke heads wounded during the manufacturing process. A full-length cDNA encoding for a putative polyphenol oxidase (designated as CsPPO) along with a 1432 bp sequence upstream of the starting ATG codon was characterized for the first time from [Cynara cardunculus var. scolymus (L.) Fiori]. The 1764 bp CsPPO sequence encodes a putative protein of 587 amino acids with a calculated molecular mass of 65,327 Da and an isoelectric point of 5.50. Analysis of the promoter region revealed the presence of cis-acting elements, some of which are putatively involved in the response to light and wounds. Expression analysis of the gene in wounded capitula indicated that CsPPO was significantly induced after 48 h, even though the browning process had started earlier. This suggests that the early browning event observed in artichoke heads was not directly related to de novo mRNA synthesis. Finally, we provide the complete gene sequence encoding for polyphenol oxidase and the upstream regulative region in artichoke.

摘要

多酚氧化酶(PPO)酶可以催化酚类物质氧化为醌类物质,据报道,它参与了许多植物性食物中不良褐变的过程。这种现象在朝鲜蓟头部在加工过程中受伤时尤为严重。首次从[Cynara cardunculus var. scolymus(L.)Fiori]中鉴定出全长编码假定多酚氧化酶(命名为 CsPPO)的 cDNA,以及起始 ATG 密码子上游的 1432 bp 序列。1764 bp 的 CsPPO 序列编码一个假定的 587 个氨基酸的蛋白质,计算的分子量为 65327 Da,等电点为 5.50。启动子区域分析显示存在顺式作用元件,其中一些可能参与对光和伤口的反应。对受伤的头状花序中的基因表达分析表明,CsPPO 在 48 小时后显著诱导,尽管褐变过程更早开始。这表明在朝鲜蓟头部观察到的早期褐变事件与新的 mRNA 合成没有直接关系。最后,我们提供了编码多酚氧化酶的完整基因序列和朝鲜蓟的上游调控区。

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