Monoterpene biosynthesis in lemon (Citrus limon) - cDNA isolation and functional analysis of four monoterpene synthases

Lücker, Joost, El Tamer, Mazen K., Schwab, Wilfried, Verstappen, Francel W. A., van der Plas, Linus H. W., Bouwmeester, Harro J. and Verhoeven, Harrie A. (2002) Monoterpene biosynthesis in lemon (Citrus limon) - cDNA isolation and functional analysis of four monoterpene synthases. European Journal of Biochemistry, 269 13: 3160-3171. doi:10.1046/j.1432-1033.2002.02985.x


Author Lücker, Joost
El Tamer, Mazen K.
Schwab, Wilfried
Verstappen, Francel W. A.
van der Plas, Linus H. W.
Bouwmeester, Harro J.
Verhoeven, Harrie A.
Title Monoterpene biosynthesis in lemon (Citrus limon) - cDNA isolation and functional analysis of four monoterpene synthases
Formatted title
Monoterpene biosynthesis in lemon (Citrus limon) - cDNA isolation and functional analysis of four monoterpene synthases
Journal name European Journal of Biochemistry   Check publisher's open access policy
ISSN 0014-2956
Publication date 2002-07-01
Sub-type Article (original research)
DOI 10.1046/j.1432-1033.2002.02985.x
Open Access Status Not yet assessed
Volume 269
Issue 13
Start page 3160
End page 3171
Total pages 12
Language eng
Abstract Citrus limon possesses a high content and large variety of monoterpenoids, especially in the glands of the fruit flavedo. The genes responsible for the production of these monoterpenes have never been isolated. By applying a random sequencing approach to a cDNA library from mRNA isolated from the peel of young developing fruit, four monoterpene synthase cDNAs were isolated that appear to be new members of the previously reported tpsb family. Based on sequence homology and phylogenetic analysis, these sequences cluster in two separate groups. All four cDNAs could be functionally expressed in Escherichia coli after removal of their plastid targeting signals. The main products of the enzymes in assays with geranyl diphosphate as substrate were (+)-limonene (two cDNAs) (–)-β-pinene and γ-terpinene. All enzymes exhibited a pH optimum around 7; addition of Mn2+ as bivalent metal ion cofactor resulted in higher activity than Mg2+, with an optimum concentration of 0.6 mm. Km values ranged from 0.7 to 3.1 µm. The four enzymes account for the production of 10 out of the 17 monoterpene skeletons commonly observed in lemon peel oil, corresponding to more than 90% of the main components present.
Keyword Citrus limon
Functional expression
(+)-limonene synthase
(-)-beta-pinene synthase
gamma-terpinene synthase
Fir Abies-Grandis
Diphosphate Synthase
Glandular Trichomes
Molecular-Cloning
Linalool Synthase
Crystal-Structure
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown
Additional Notes Journal title continues (2005-) The FEBS Journal (1742-464X; 1742-4658)

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Biological Sciences Publications
 
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