Enzymes and methods for producing omega-3 fatty acids
The present invention relates generally to the field of recombinant fatty acid synthesis, particularly in transgenic plants. The application describes genes involved in fatty acid synthesis and provides methods and vectors for the manipulation of fatty acid composition of plant oils. In particular, the invention provides constructs for achieving the integration of multiple heterologous genes involved in fatty acid synthesis into the plant genome, such that the resulting plants produce altered levels of polyunsaturated fatty acids. Also described are methods for enhancing the expression of fatty acid biosynthesis enzymes by co-expressing a silencing suppressor within the plant storage organ.
1. A process for producing seed comprising eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, the process comprising the steps of:
(i) growing a Brassica napus plant comprising exogenous polynucleotides encoding a microalgal Δ6-desaturase which has greater Δ6-desaturase activity on ALA-CoA as fatty acid substrate than on ALA joined to the sn-2 position of phosphatidylcholine (PC) as the fatty acid substrate, a Δ6-elongase, a microalgal Δ5-desaturase, a microalgal Δ5-elongase, and a microalgal Δ4-desaturase, wherein each polynucleotide is operably linked to one or more promoters that direct expression of said polynucleotides during the development of the seed, to produce seed that comprises eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid which were produced from α-linolenic acid (ALA) during development of the seed and which are present in an esterified form as part of triglycerides at levels based on an efficiency of conversion of α-linolenic acid to eicosapentaenoic acid, docosapentaenoic acid or docosahexaenoic acid of at least 17.3% and an efficiency of conversion of eicosapentaenoic acid to docosapentaenoic acid of at least 60%, and (ii) harvesting the seed from the plant.
2. The process of claim 1 , wherein docosapentaenoic acid and docosahexaenoic acid are present at levels based on an efficiency of conversion of α-linolenic acid to docosapentaenoic acid or docosahexaenoic acid of at least 15.4%.
3. The process of claim 1 , wherein docosahexaenoic acid is present at a level based on an efficiency of conversion of α-linolenic acid to docosahexaenoic acid of at least 9.5%.
4. The process of claim 1 , wherein docosahexaenoic acid is present at a level based on an efficiency of conversion of eicosapentaenoic acid to docosahexaenoic acid of at least 45%.
5. The process of claim 1 , wherein arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid together comprise 15% of the total fatty acids in the seed.
6. The process of claim 1 , wherein the Δ6-desaturase is an Ostreococcus tauri or Micromonas pusilla Δ 6-desaturase.