Cloning, functional characterization and genomic organization of 1,8-cineole synthases from Lavandula
department or school: Arts and Sciences, Irving K. Barber School of (Okanagan)
department or school: Biology, Department of (Okanagan)
data provider: DSpace
identifier citation: Demissie ZA, Cella M, Sarker L, Thompson T, Rheault M, and Mahmoud SS (2012). Cloning, functional characterization and genomic organization of 1,8-cineole synthases from Lavandula. Plant Molecular Biology. 79(4): 393-411.
creator: Demissie, Zerihun Abebe
creator: Cella, Monica Angela
creator: Sarker, Lukman S.
creator: Thompson, Travis J.
creator: Rheault, Mark
creator: Mahmoud, Soheil S.
issued: 2019-01-08T17:21:00Z
issued: 2012
description: Several members of the genus Lavandula produce valuable essential oils (EOs) that are primarily constituted of the low molecular weight isoprenoids, particularly monoterpenes. We isolated over 8,000 ESTs from the glandular trichomes of L. x intermedia flowers (where bulk of the EO is synthesized) to facilitate the discovery of genes that control the biosynthesis of EO constituents. The expression profile of these ESTs in L. x intermedia and its parents L. angustifolia and L. latifolia was established using microarrays. The resulting data highlighted a differentially expressed, previously uncharacterized cDNA with strong homology to known 1,8-cineole synthase (CINS) genes. The ORF, excluding the transit peptide, of this cDNA was expressed in E. coli, purified by Ni-NTA agarose affinity chromatography and functionally characterized in vitro. The ca. 63 kDa bacterially produced recombinant protein, designated L. x intermedia CINS (LiCINS), converted geranyl diphosphate (the linear monoterpene precursor) primarily to 1,8-cineole with K ( m ) and k ( cat ) values of 5.75 μM and 8.8 × 10-³ s-¹, respectively. The genomic DNA of CINS in the studied Lavandula species had identical exon-intron architecture and coding sequences, except for a single polymorphic nucleotide in the L. angustifolia ortholog which did not alter protein function. Additional nucleotide variations restricted to L. angustifolia introns were also observed, suggesting that LiCINS was most likely inherited from L. latifolia. The LiCINS mRNA levels paralleled the 1,8-cineole content in mature flowers of the three lavender species, and in developmental stages of L. x intermedia inflorescence indicating that the production of 1,8 cineole in Lavandula is most likely controlled through transcriptional regulation of LiCINS. [This is a post-peer-review, pre-copyedit version of an article published in Plant Molecular Biology. The final authenticated version is available online at: http://dx.doi.org/10.1007/s11103-…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/68205
note: 1 Cloning, functional characterization and genomic organization of 1,8-cineole synthases from Lavandula Zerihun A. Demissie • Monica A. Cella • Lukman S. Sarker • Travis J. Thompson • Mark R. Rheault • Soheil S. Mahmoud Department of Biology, University of British Columbia, 1177 Research Road, Kelowna, British Columbia V1V 1V7, Canada Corresponding author e-mail: soheil.mahmoud@ubc.ca Abstract Several members of the genus Lavandula produce valuable essential oils (EOs) that are primarily constituted of the low molecular weight isoprenoids, particularly monoterpenes. We isolated over 8,000 ESTs from the glandular trichomes of L. x intermedia flowers (where bulk of the EO is synthesized) to facilitate the discovery of genes that control the biosynthesis of EO constituents. The expression profile of these ESTs in L. x intermedia and its parents L. angustifolia and L. latifolia was established using microarrays. The resulting data highlighted a differentially expressed, previously uncharacterized cDNA with strong homology to known 1,8-cineole synthase (CINS) genes. The ORF, excluding the transit peptide, of this cDNA was expressed in E. coli, purified by Ni-NTA agarose affinity chromatography and functionally characterized in vitro. The ca. 63 kDa bacterially produced recombinant protein, designated L. x intermedia CINS (LiCINS), converted geranyl diphosphate (the linear monoterpene precursor) primarily to 1,8-cineole with Km and kcat values of 5.75 µM and 8.8 x 10-3 s-1, respectively. The genomic DNA of CINS in the studied Lavandula species had identical exon-intron architecture and coding sequences, except for a single polymorphic nucleotide in the L. angustifolia ortholog which did not alter protein function. Additional nucleotide variations restricted to L. angustifolia introns were also observed, suggesting that LiCINS was most likely inherited from L. latifolia. The LiCINS mRNA levels paralleled the 1,8-cineole content in mature flowers of the three lavender speci…
has type: Article
has type: Postprint
is shown at: 10.14288/1.0375951
language: eng
peer review status: Reviewed
provider: Vancouver : University of British Columbia Library
publisher DOI: 10.1007/s11103-012-9920-3
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Faculty
subject: L. x intermedia
subject: L. angustifolia
subject: L. latifolia
subject: Essential oil
subject: Isoprenoid
type: Text
identifier URI: http://hdl.handle.net/2429/68205