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Isolation and Characterisation of a High-Efficiency Desaturase and Elongases from Microalgae for Transgenic LC-PUFA Production

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Abstract

The production of long-chain polyunsaturated fatty acids from precursor molecules linoleic acid (LA; 18:2ω6) and α-linolenic acid (ALA; 18:3ω3) is catalysed by sequential desaturase and elongase reactions. We report the isolation of a front-end Δ6-desaturase gene from the microalgae Ostreococcus lucimarinus and two elongase genes, a Δ6-elongase and a Δ5-elongase, from the microalga Pyramimonas cordata. These enzymes efficiently convert their respective substrates when transformed in yeast (39–75% conversion for ω3 substrate fatty acids), and the Δ5-elongase in particular displays higher elongation efficiency (75% for conversion of eicosapentaenoic acid (20:5ω3) to docosapentaenoic acid (22:5ω3)) than previously reported genes. In addition, the Δ6-desaturase is homologous with acyl-CoA desaturases and shows a strong preference for the ω3 substrate ALA.

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Acknowledgements

We gratefully acknowledge the excellent technical assistance provided by Rob Gurney during yeast feeding experiments and Adam White during construct building and yeast transformation. This research was funded by the Food Futures Flagship and the Commonwealth Scientific and Industrial Research Organisation. Two anonymous reviewers and Dr Grant Burgess are thanked for their comments, which improved the manuscript.

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Authors and Affiliations

  1. Food Futures National Research Flagship, CSIRO Plant Industry, P. O. Box 1600, Canberra, ACT, 2601, Australia

    James R. Petrie, Qing Liu, Anne M. Mackenzie, Pushkar Shrestha & Surinder P. Singh

  2. Food Futures National Research Flagship, CSIRO Marine and Atmospheric Research, P. O. Box 1538, Hobart, TAS, 7000, Australia

    Maged P. Mansour, Stan S. Robert, Dion F. Frampton, Susan I. Blackburn & Peter D. Nichols

Authors
  1. James R. Petrie
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  2. Qing Liu
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  3. Anne M. Mackenzie
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  4. Pushkar Shrestha
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  5. Maged P. Mansour
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  6. Stan S. Robert
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  7. Dion F. Frampton
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  8. Susan I. Blackburn
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  9. Peter D. Nichols
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  10. Surinder P. Singh
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Corresponding author

Correspondence to Surinder P. Singh.

Additional information

Genbank Accessions: O. lucimarinus Δ6-desaturase = BK006814; P. cordata Δ6-elongase = GQ202034; P. cordata Δ5-elongase = GQ202035.

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Petrie, J.R., Liu, Q., Mackenzie, A.M. et al. Isolation and Characterisation of a High-Efficiency Desaturase and Elongases from Microalgae for Transgenic LC-PUFA Production. Mar Biotechnol 12, 430–438 (2010). https://doi.org/10.1007/s10126-009-9230-1

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  • DOI: https://doi.org/10.1007/s10126-009-9230-1

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