Molecular cloning and characterization of a Perilla frutescens cytochrome P450 enzyme that catalyzes the later steps of perillaldehyde biosynthesis
Molecular cloning and characterization of a Perilla frutescens cytochrome P450 enzyme that catalyzes the later steps of perillaldehyde biosynthesis
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DOI：
10.1016/j.phytochem.2016.11.009
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发表时间：
2017-02-01
期刊：
影响因子：
3.8
通讯作者：
Ito, Michiho
中科院分区：
文献类型：
--
作者：
Fujiwara, Yumi;Ito, Michiho
紫苏产生环己烷单萜紫苏醛，作为精油的主要成分，在其腺毛中积累。紫苏醛是大豆质量控制的标记化合物，具有抗菌、镇静或血管舒张等生物活性。预测的紫苏醛形成涉及香叶基二磷酸的环化、羟基化和氧化，细胞色素P450在紫苏醛生物合成中起着至关重要的作用。本研究通过分析几种紫苏纯系油品的表达序列标签库，分离出具有紫苏醇和紫苏醛合酶活性的细胞色素P450型酶。具有对紫苏醛类型具有高度特异性的序列的重组蛋白在酿酒酵母中表达，并通过体外酶促反应进行评估。重组蛋白催化柠檬烯羟基化和氧化生成紫苏醇和紫苏醛。先前已从紫苏中分离出细胞色素 P450 柠檬烯-7-羟化酶 cDNA。本研究中分离的细胞色素 P450 与之前分离的酶有 37% 的氨基酸同一性；然而，它可能具有不同的特征。 (C) 2016 Elsevier Ltd. 保留所有权利。
Perilla produces the cyclohexanoid monoterpene perillaldehyde as a major constituent of an essential oil that is accumulated in its glandular trichomes. Perillaldehyde is a marker compound for quality control of soyo and has biological activities such as antibacterial, sedative, or vasodilatory effects. The predicted perillaldehyde formation involves the cyclization of geranyl diphosphate, hydroxylation, and oxidation, and cytochrome P450 plays a crucial role in perillaldehyde biosynthesis. In this study, a cytochrome P450-type enzyme with perillyl alcohol and perillaldehyde synthase activities was isolated by analyzing an expressed sequence tag library from several oil types of pure lines of perilla. A recombinant protein with a sequence that was highly specific for the type of perillaldehyde was expressed in Saccharomyces cerevisiae and evaluated by an in vitro enzymatic reaction. The recombinant protein catalyzed the hydroxylation and oxidation of limonene to perillyl alcohol and perillaldehyde. Cytochrome P450 limonene-7-hydroxylase cDNA from Perilla frutescens has been previously isolated. The cytochrome P450 isolated in this study shares 37% amino-acid identity with the previously isolated enzyme; however, it may have different characteristics. (C) 2016 Elsevier Ltd. All rights reserved.