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Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq

BACKGROUND: 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 t...

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Autores principales: Engprasert, Surang, Taura, Futoshi, Kawamukai, Makoto, Shoyama, Yukihiro
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC538753/
https://www.ncbi.nlm.nih.gov/pubmed/15550168
http://dx.doi.org/10.1186/1471-2229-4-18
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author Engprasert, Surang
Taura, Futoshi
Kawamukai, Makoto
Shoyama, Yukihiro
author_facet Engprasert, Surang
Taura, Futoshi
Kawamukai, Makoto
Shoyama, Yukihiro
author_sort Engprasert, Surang
collection PubMed
description BACKGROUND: 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. RESULTS: 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. CONCLUSION: 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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spelling pubmed-5387532004-12-22 Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq Engprasert, Surang Taura, Futoshi Kawamukai, Makoto Shoyama, Yukihiro BMC Plant Biol Research Article BACKGROUND: 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. RESULTS: 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. CONCLUSION: 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. BioMed Central 2004-11-18 /pmc/articles/PMC538753/ /pubmed/15550168 http://dx.doi.org/10.1186/1471-2229-4-18 Text en Copyright © 2004 Engprasert et al; licensee BioMed Central Ltd.
spellingShingle Research Article
Engprasert, Surang
Taura, Futoshi
Kawamukai, Makoto
Shoyama, Yukihiro
Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title_full Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title_fullStr Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title_full_unstemmed Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title_short Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq
title_sort molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from coleus forskohlii briq
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC538753/
https://www.ncbi.nlm.nih.gov/pubmed/15550168
http://dx.doi.org/10.1186/1471-2229-4-18
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