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Molecular Cloning and Functional Characterization of Fatty Acyl Desaturase and Elongase cDNAs Involved in the Production of Eicosapentaenoic and Docosahexaenoic Acids from α-Linolenic Acid in Atlantic Salmon (Salmo salar)

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Abstract

Fish are the only major dietary source for humans of ω-3 highly unsaturated fatty acids (HUFAs) and with declining fisheries farmed fish such as Atlantic salmon (Salmo salar) constitute an increasing proportion of the fish in the human diet. However, the current high use of fish oils, derived from wild capture marine fisheries, in aquaculture feeds is not sustainable in the longer term and will constrain continuing growth of aquaculture activities. Greater understanding of how fish metabolize and biosynthesize HUFA may lead to more sustainable aquaculture diets. The study described here contributes to an effort to determine the molecular genetics of the HUFA biosynthetic pathway in salmon, with the overall aim being to determine mechanisms for optimizing the use of vegetable oils in Atlantic salmon culture. In this paper we describe the cloning and functional characterization of 2 genes from salmon involved in the biosynthesis of HUFA. A salmon desaturase complementary DNA, SalDes, was isolated that include an open reading frame of 1362 bp specifying a protein of 454 amino acids. The protein sequence includes all the characteristics of microsomal fatty acid desaturases, including 3 histidine boxes, 2 transmembrane regions, and an N-terminal cytochrome b5 domain containing a heme-binding motif similar to that of other fatty acid desaturases. Functional expression in the yeast Saccharomyces cerevisiae showed SalDes is predominantly an ω-3 δ5 desaturase, a key enzyme in the synthesis of eicosapentaenoic acid (20:5n-3) from α-linolenic acid (18:3n-3). The desaturase showed only low levels of δ6 activity toward C18 polyunsaturated fatty acids. In addition, a fatty acid elongase cDNA, SalElo, was isolated that included an open reading frame of 888 bp, specifying a protein of 295 amino acids. The protein sequence of SalElo included characteristics of microsomal fatty acid elongases, including a histidine box and a transmembrane region. Upon expression in yeast SalElo showed broad substrate specificity for polyunsaturated fatty acids with a range of chain lengths, with the rank order being C18 > C20 > C22. Thus this one polypeptide product displays all fatty acid elongase activities required for the biosynthesis of docosahexaenoic acid (22:6n-3) from 18:3n-3.

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Acknowledgments

This work was supported partly by a Scottish Higher Education Funding Council (SHEFC) Grant (“Aquagene”) and partly by the European Union Framework V Programme (project QLRT-2000-30058, “RAFOA”). N.H. was in receipt of a Natural Environment Research Council studentship and gratefully acknowledges the additional support of an award from the Scottish International Education Trust (SIET).

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  1. Institute of Aquaculture, University of Stirling, FK9 4LA, Scotland, U.K.

    Nicola Hastings, Morris K. Agaba, Douglas R. Tocher, Xiaozhong Zheng, Cathryn A. Dickson, James R. Dick & Alan J. Teale

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  1. Nicola Hastings
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  2. Morris K. Agaba
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  3. Douglas R. Tocher
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  4. Xiaozhong Zheng
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Correspondence to Douglas R. Tocher.

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Hastings, N., Agaba, M.K., Tocher, D.R. et al. Molecular Cloning and Functional Characterization of Fatty Acyl Desaturase and Elongase cDNAs Involved in the Production of Eicosapentaenoic and Docosahexaenoic Acids from α-Linolenic Acid in Atlantic Salmon (Salmo salar). Mar Biotechnol 6, 463–474 (2004). https://doi.org/10.1007/s10126-004-3002-8

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