Isolation and characterization of a cDNA encoding a delta8 sphingolipid desaturase from Aquilegia vulgaris

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Michaelson, L. V., Longman, A. J., Sayanova, O. V., Stobart, A. K. and Napier, J. A. 2002. Isolation and characterization of a cDNA encoding a delta8 sphingolipid desaturase from Aquilegia vulgaris. Biochemical Society Transactions. 30 (6), pp. 1073-1075.

AuthorsMichaelson, L. V., Longman, A. J., Sayanova, O. V., Stobart, A. K. and Napier, J. A.
Abstract

We have isolated a cDNA encoding the Delta(8) sphingolipid desaturase from the plant Aquilegia vulgaris L. via a PCR-based strategy using primers designed to target the conserved histidine box regions of microsomal desaturases. The function of the cDNA was confirmed by expression in the yeast, Saccharomyces cerevisiae. Analysis of the long-chain sphingoid bases as their dinitrophenyl derivatives by reverse-phase HPLC demonstrated the accumulation of cis - and trans -desaturated sphingoid bases which were not present in the wild-type yeast cells. The Delta(8) desaturated products co-eluted with known Delta(8)-desaturated phytosphingenine and the molecular mass of these products was confirmed by liquid chromatography-MS. The Delta(8) long-chain base desaturase was also able to desaturate dihydrosphingosine substrates. This is the first report of the functional characterization of an A. vulgaris gene product.

Year of Publication2002
JournalBiochemical Society Transactions
Journal citation30 (6), pp. 1073-1075
Digital Object Identifier (DOI)doi:10.1042/bst0301073
PubMed ID12440975
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or code415
503
Isolation of DNAs for novel fatty acid desaturase enzymes and their use to manipulate fatty acid and triglycerol composition in transgenic plants
Output statusPublished
Publication dates
Online01 Nov 2002
Copyright licensePublisher copyright
ISSN0300-5127
PublisherPortland Press Ltd

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