Journal of the American Oil Chemists' Society

, Volume 84, Issue 9, pp 809–816

Cloning of Flax Oleic Fatty Acid Desaturase and Its Expression in Yeast

  • Anna Krasowska
  • Dorota Dziadkowiec
  • Anna Polinceusz
  • Aneta Plonka
  • Marcin Łukaszewicz
Original Paper

Abstract

Fatty acid desaturases dehydrogenate acyl chains, which results in the formation of a double bond. Using PCR on flax genomic DNA, we cloned a putative Δ12 fatty acid desaturase (Fad2) gene encoding a 378 amino acid protein. Heterologous expression of this protein in yeast as an N-terminal fusion to GFP showed its localization within endoplasmic reticulum. Analysis of membrane lipids revealed the production of dienoic fatty acids, decreased levels of FAD2 substrates and an increased concentration of longer fatty acids. Higher peroxidation of lipids in FAD2-containing strains is not reflected by any visible phenotype in YPD medium. However, FAD2-containing strains with deleted superoxide dismutase genes exhibited significant growth reductions under oxidative stress.

Keywords

FAD2 fatty acid desaturase Linum usitatissimum Saccharomyces cerevisiae Cu–Zn superoxide dismutase Manganese superoxide dismutase Omega-6 fatty acids 

Abbreviations

GFP

Green fluorescent protein

FAD2

Oleic acid (Δ12) fatty acid desaturase

FA

Fatty acid

PUFA

Polyunsaturated fatty acid

TAG

Triacylglycerols

SOD

Superoxide dismutase

FAMEs

Fatty acid methyl esters

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Copyright information

© AOCS 2007

Authors and Affiliations

  • Anna Krasowska
    • 1
  • Dorota Dziadkowiec
    • 1
  • Anna Polinceusz
    • 1
  • Aneta Plonka
    • 1
  • Marcin Łukaszewicz
    • 1
  1. 1.Faculty of BiotechnologyWrocław UniversityPrzybyszewskiegoPoland

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