Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce
Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce
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DOI：
10.1016/j.phytochem.2006.09.016
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发表时间：
2006-12-01
期刊：
影响因子：
3.8
通讯作者：
Ebizuka, Yutaka
中科院分区：
文献类型：
--
作者：
Basyuni, Mohammad;Oku, Hirosuke;Ebizuka, Yutaka
根据已知氧化角鲨烯环化酶（oxidosqualene cylases，OSC）的保守序列设计简并引物，进行同源性PCR，从秋茄幼根中克隆了一个三萜合酶（triterpenesynthase，KcMS）基因。Druce（Rhizophoraceae）. KcMS由一个2286 bp的开放阅读框架组成，编码761个氨基酸。推导的氨基酸序列与蓖麻羽扇豆醇合成酶的氨基酸序列同源性为79%，表明该酶为秋茄羽扇豆醇合成酶。利用GAL 1启动子控制的表达载体pYES 2在羊毛甾醇合成酶缺陷型酵母中表达KcMS。GC-MS分析表明，KcMS中羽扇豆醇、β-香树脂醇和α-香树脂醇的比例为2：1：1，表明KcMS编码一个多功能三萜合酶。羽扇豆醇合酶这是第一个从红树林中克隆的OSC。(c)2006爱思唯尔有限公司版权所有。
Homology based PCRs with degenerate primers designed from the conserved sequences among the known oxidosqualene cylases (OSCs) have resulted in cloning of a triterpene synthase (KcMS) from the young roots of Kandelia candel (L.) Druce (Rhizophoraceae). KcMS consists of a 2286 bp open reading frame, which codes for 761 amino acids. The deduced amino acid sequence showed 79% homology to a lupeol synthase from Ricinus communis suggesting it to be a lupeol synthase of K candel. KcMS was expressed in a lanosterol synthase deficient yeast with the expression vector pYES2 under the control of GAL1 promoter. GC-MS analysis showed that the transformant accumulated a mixture of lupeol, P-amyrin and alpha-amyrin in a 2:1:1 ratio, indicating that KcMS encodes a multifunctional triterpene synthase, although it showed high sequence homology to a R. communis lupeol synthase. This is the first OSC cloning from mangrove tree species. (c) 2006 Elsevier Ltd. All rights reserved.