Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba.
Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba.
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银杏萜三内酯生物合成中乙酰辅酶A C-乙酰转移酶和甲羟戊酸激酶基因的分子克隆、表征和功能分析
DOI：
10.3390/molecules22010074
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发表时间：
2017-01-02
期刊：
影响因子：
--
通讯作者：
Qu J
中科院分区：
文献类型：
--
作者：
Chen Q;Yan J;Meng X;Xu F;Zhang W;Liao Y;Qu J
银杏内酯和白果内酯，统称为萜三内酯（TTL），是萜类化合物，形成银杏的主要活性物质。甲羟戊酸（MVA）生物合成途径中的萜类化合物包括乙酰辅酶A C-乙酰转移酶（AACT）和甲羟戊酸激酶（MVK）作为核心酶。本研究从中国对虾中克隆了AACT（GbAACT，GenBank Accession No.KX904942）和MVK（GbMVK，GenBank Accession No.KX904944）的全长cDNA。biloba. GbAACT和GbMVK蛋白分别由404和396个氨基酸组成，开放阅读框（ORF）分别为1215 bp和1194 bp。组织表达谱分析表明，GbAACT在银杏果实和叶片中高表达，GbMVK在叶片和根中高表达。GbAACT在AACT缺陷型酿酒酵母菌株Δ erg 10中的功能互补和GbMVK在MVK缺陷型菌株Δ erg 12中的功能互补证实了GbAACT介导甲羟戊酸乙酰辅酶A转化为乙酰乙酰辅酶A，GbMVK介导甲羟戊酸转化为甲羟戊酸磷酸。这一观察结果表明，GbAACT和GbMVK是细胞溶质甲羟戊酸（MVA）生物合成途径中的功能基因。继G.茉莉酸甲酯和水杨酸处理银杏幼苗后，GbAACT和GbMVK的表达量增加，TTL产量增加。GbAACT和GbMVK基因的克隆、鉴定、表达和功能分析将有助于进一步了解这两个基因在TTL生物合成中的作用。
Ginkgolides and bilobalide, collectively termed terpene trilactones (TTLs), are terpenoids that form the main active substance of Ginkgo biloba. Terpenoids in the mevalonate (MVA) biosynthetic pathway include acetyl-CoA C-acetyltransferase (AACT) and mevalonate kinase (MVK) as core enzymes. In this study, two full-length (cDNAs) encoding AACT (GbAACT, GenBank Accession No. KX904942) and MVK (GbMVK, GenBank Accession No. KX904944) were cloned from G. biloba. The deduced GbAACT and GbMVK proteins contain 404 and 396 amino acids with the corresponding open-reading frame (ORF) sizes of 1215 bp and 1194 bp, respectively. Tissue expression pattern analysis revealed that GbAACT was highly expressed in ginkgo fruits and leaves, and GbMVK was highly expressed in leaves and roots. The functional complementation of GbAACT in AACT-deficient Saccharomyces cerevisiae strain Δerg10 and GbMVK in MVK-deficient strain Δerg12 confirmed that GbAACT mediated the conversion of mevalonate acetyl-CoA to acetoacetyl-CoA and GbMVK mediated the conversion of mevalonate to mevalonate phosphate. This observation indicated that GbAACT and GbMVK are functional genes in the cytosolic mevalonate (MVA) biosynthesis pathway. After G. biloba seedlings were treated with methyl jasmonate and salicylic acid, the expression levels of GbAACT and GbMVK increased, and TTL production was enhanced. The cloning, characterization, expression and functional analysis of GbAACT and GbMVK will be helpful to understand more about the role of these two genes involved in TTL biosynthesis.
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影响因子：
4.3
作者：
Kim, Sang-Min;Kuzuyama, Tomohisa;Kim, Soo-Un
通讯作者：
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影响因子：
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影响因子：
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