Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq.
Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq.
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DOI：
10.1186/1471-2229-4-18
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发表时间：
2004-11-18
影响因子：
5.3
通讯作者：
Shoyama Y
中科院分区：
文献类型：
--
作者：
Engprasert S;Taura F;Kawamukai M;Shoyama Y
异戊烯基二磷酸(IPP)是拉布丹二萜Forsklin的一种常见的生物合成前体，通过非甲氧戊酸途径进行生物合成。香叶基香叶素二磷酸合成酶(GGPP)是萜类化合物生物合成的重要分支酶。因此，GGPP合酶被认为是Forskolin生物合成的关键酶。在此，我们首次报道了甘草杆菌GGPP合成酶基因的确认。全长GGPP合成酶的开放阅读框编码一个359个氨基酸的蛋白质，其中1077个核苷酸长，计算的相对分子质量为39.3 kDa。GGPP合成酶的氨基酸序列与其他植物GGPP合成酶有很高的同源性。确定了几个高度保守的区域，包括两个富含天冬氨酸的基序。GGPP合成酶-GFP融合蛋白N-末端片段在烟草细胞中的瞬时表达证明其定位于叶绿体中。在含有尿毒欧文氏菌pACCAR25GGPP合成酶和福氏弧菌Δ合成酶的大肠杆菌中观察到类胡萝卜素的产生。这些结果表明，该基因编码功能性GGPP合成酶。此外，Forskohlii GGPP合成酶在叶片中表达较强，在茎中表达减弱，在根中表达很少。本研究提出福司可林是通过非甲氧戊酸途径合成的。GGPP合酶被认为参与了Forskolin的生物合成，Forskolin主要在叶中合成，随后在茎和根中积累。
Isopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq. The open reading frame for full-length GGPP synthase encodes a protein of 359 amino acids, in which 1,077 nucleotides long with calculated molecular mass of 39.3 kDa. Alignments of C. forskohlii GGPP synthase amino acid sequences revealed high homologies with other plant GGPP synthases. Several highly conserved regions, including two aspartate-rich motifs were identified. Transient expression of the N-terminal region of C. forskohlii GGPP synthase-GFP fusion protein in tobacco cells demonstrated subcellular localization in the chloroplast. Carotenoid production was observed in Escherichia coli harboring pACCAR25ΔcrtE from Erwinia uredovora and plasmid carrying C. forskohlii GGPP synthase. These results suggested that cDNA encoded functional GGPP synthase. Furthermore, C. forskohlii GGPP synthase expression was strong in leaves, decreased in stems and very little expression was observed in roots. This investigation proposed that forskolin was synthesised via a non-mevalonate pathway. GGPP synthase is thought to be involved in the biosynthesis of forskolin, which is primarily synthesised in the leaves and subsequently accumulates in the stems and roots.
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影响因子：
4.3
作者：
Oh, SK;Kim, IJ;Han, KH
通讯作者：
Han, KH
影响因子：
9.2
作者：
Chiu, WL;Niwa, Y;Sheen, J
通讯作者：
Sheen, J
DOI：
10.1016/1011-1344(93)80071-g
发表时间：
1993-05-01
影响因子：
5.4
作者：
SANDMANN, G;MISAWA, N;MACINO, G
通讯作者：
MACINO, G
影响因子：
1.8
作者：
BHAT, SV;BAJWA, BS;FEHLHABER, HW
通讯作者：
FEHLHABER, HW
影响因子：
7.4
作者：
Okada, K;Saito, T;Kamiya, Y
通讯作者：
Kamiya, Y