A Sphingolipid Desaturase from Higher Plants

  title={A Sphingolipid Desaturase from Higher Plants},
  author={Petra Sperling and Ulrich Zähringer and Ernst Heinz},
  journal={The Journal of Biological Chemistry},
  pages={28590 - 28596}
A recently cloned cDNA from sunflower codes for a fusion protein composed of an N-terminal cytochromeb 5 and a domain similar to membrane-bound acyl lipid desaturases. For a functional identification, homologous cDNAs from Brassica napus and Arabidopsis thaliana were expressed in Saccharomyces cerevisiae, and sphingolipid long chain bases were analyzed. The expression of the heterologous enzyme results in significant proportions of new Δ8,9-cis/trans-phytosphingenines that accompany the… 

Figures from this paper

phingolipid base modifying enzymes in sunflower ( Helianthus annuus ) : Cloning nd characterization of a C 4-hydroxylase gene and a new paralogous 8-desaturase gene ntonio
A C4-long chain base hydroxylase was functionally characterized in sunflower plant, an enzyme that could complement the sur2 mutation when heterologously expressed and be important for complex sphingolipids finally becoming functional.
Functional identification of a delta8-sphingolipid desaturase from Borago officinalis.
The similarities between delta12- and delta5-fatty acyl desaturase sequences were used to construct degenerate primers for PCR experiments with cDNA transcribed from mRNA of developing borage seeds and showed high similarity to plant delta8-sphingolipid desaturases.
A bifunctional Δ6‐fatty acyl acetylenase/desaturase from the moss Ceratodon purpureus
It is demonstrated that the Delta6-acetylenase from C. pupureus is a bifunctional enzyme, which can introduce a Delta6cis-double bond into 9,12,(15)-C18-polyenoic acids as well as converting a Delta8-sphingolipid bond to a Delta 6-triple bond.
Genes encoding Δ8-sphingolipid desaturase from various plants: identification, biochemical functions, and evolution
The genes of 20 ∆8-sphingolipid desaturases from 12 plants were identified and functionally detected by using Saccharomyces cerevisiae system to elucidate the relationship between the biochemical function and evolution of this enzyme.
Heterologous expression of Mucor rouxii delta(12)-desaturase gene in Saccharomyces cerevisiae.
A gene whose product is responsible for Delta(12)-desaturase activity and is involved in the metabolic pathway of gamma-linolenic acid (GLA) synthesis of Mucor rouxii is presented and shows similarity with microsomal omega-3 and omega-6 desaturases of plants.
Cloning and molecular characterization of the Δ6-desaturase from two Echium plant species: Production of GLA by heterologous expression in yeast and tobacco
Cloned genes encoding the Δ6-desaturase (D6DES) from two different Macaronesian Echium species, E. pitardii and E. gentianoides, show a positive correlation between the D6DES transcript level and GLA accumulation in different tissues of the plant, and biotechnological application can be envisaged as an initial step toward the generation of transgenic oleaginous plants producing GLA.


Identification of a Caenorhabditis elegans Δ6-fatty-acid-desaturase by heterologous expression in Saccharomyces cerevisiae
This work identified a cDNA expressed sequence tag from an animal (the nematode worm Caenorhabditis elegans) that showed weak similarity to a higher-plant microsomal Delta6-desaturase, and isolated and expressed the protein, which demonstrated that the protein encoded by the C. elegans cDNA was that of a fatty acid Delta6, as determined by the accumulation of gamma-linolenic acid.
Identification of a novel delta 6-acyl-group desaturase by targeted gene disruption in Physcomitrella patens.
This is the first report of a successful gene disruption in a multicellular plant resulting in a specific biochemical phenotype, and the new cDNA coded for a delta 6-desaturase was confirmed by expression of the cDNA in yeast and analysis of the resultant fatty acids by GC-MS.
A cytochrome-b5-containing fusion protein similar to plant acyl lipid desaturases.
The similarity between oleate and linoleate desaturase sequences from several plants was used to construct degenerate oligonucleotide primers for PCR experiments with cDNA transcribed from mRNA of ripening sunflower embryos and desaturases revealed to be novel members of the cytochrome b5 superfamily.
Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of delta6-desaturated fatty acids in transgenic tobacco.
  • O. Sayanova, M. Smith, J. Napier
  • Biology, Medicine
    Proceedings of the National Academy of Sciences of the United States of America
  • 1997
To enable the production of GLA in conventional oilseeds, a cDNA encoding the delta6-fatty acid desaturase from developing seeds of borage is isolated and its function is confirmed by expression in transgenic tobacco plants.
A Novel Cytochrome b5-like Domain Is Linked to the Carboxyl Terminus of the Saccharomyces cerevisiae Δ-9 Fatty Acid Desaturase (*)
In animal and fungal cells cytochrome b5 is thought to be an electron donor for sterol modifying enzymes and fatty acid desaturases and may play some nonessential role in these functions, however, disruption of the Saccharomyces cyto chrome b5 gene yielded cells that had no nutritional requirement for either sterols or unsaturated fatty acids.
Fah1p, a Saccharomyces cerevisiae Cytochromeb 5 Fusion Protein, and ItsArabidopsis thaliana Homolog That Lacks the Cytochromeb 5 Domain Both Function in the α-Hydroxylation of Sphingolipid-associated Very Long Chain Fatty Acids*
Analysis of the FAH1 gene in S. cerevisiaedid revealed similarities to Ole1p, which suggested that Fah1p played a role in the biosynthesis or modification of fatty acids, and also revealed a number of homologs to this gene in other species.
Desaturase multigene families of Brassica napus arose through genome duplication
Estimates of the gene copy number for the five different desaturase cDNA clones from Brassica napus supported the premise that all these genes were at least duplicated or triplicated in the two progenitor species before they combined to form B. napus, suggesting that the desatur enzyme multigene families arose as the result of duplication of large chromosome segments rather than duplication of individual genes.
Hydroxylation of Saccharomyces cerevisiae Ceramides Requires Sur2p and Scs7p*
Characterization of scs7 andsur2 mutants is expected to provide insight into the function of ceramide hydroxylation.
Polyunsaturated fatty acid biosynthesis in Saccharomyces cerevisiae: expression of ethanol tolerance and the FAD2 gene from Arabidopsis thaliana
The Arabidopsis thaliana delta-12 fatty acid desaturase gene (FAD2) was overexpressed in Saccharomyces cerevisiae by using the GAL1 promoter to assess the ethanol tolerance of this strain, and cells overexpressing the FAD2 gene had greater resistance to 15% ethanol than did the control cells.