IP Library › Granted Patent US 12,195,739
Granted Patent B2
US 12,195,739 · App. 18/409,591 · Granted Jan 14, 2025

Enzymes and methods for producing omega-3 fatty acids

Inventors: James Robertson Petrie (Bywong, AU); Anne Maree Mackenzie (Hawker, AU); Qing Liu (Giralang, AU); Pushkar Shrestha (Lawson, AU); Peter David Nichols (West Hobart, AU); Susan Irene Ellis Blackburn (Battery Point, AU); Maged Peter Mansour (Gladstone Park, AU); Stanley Suresh Robert (Oyster Cove, AU); Dion Matthew Frederick Frampton (Bonnet Hill, AU); Xue-Rong Zhou (Harrison, AU); Surinder Pal Singh (Downer, AU); Craig Christopher Wood (Dickson, AU)
Assignees: COMMONWEALTH SCIENTIFIC AND INDUSTRIAL RESEARCH ORGANISATION; GRAINS RESEARCH AND DEVELOPMENT CORPORATION
C12N15/8247C12P7/6427C12P7/6432C12P7/6434C12P7/6472C12Y602/01003
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Quick Facts
Patent No.
US 12,195,739
App. No.
18/409,591
Granted
Jan 14, 2025
Kind
B2
Abstract

The present invention relates to methods of synthesizing long-chain polyunsaturated fatty acids, especially eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, in recombinant cells such as yeast or plant cells. Also provided are recombinant cells or plants which produce long-chain polyunsaturated fatty acids. Furthermore, the present invention relates to a group of new enzymes which possess desaturase or elongase activity that can be used in methods of synthesizing long-chain polyunsaturated fatty acids. In particular, the present invention provides ω3 destaurases, Δ5 elongases and Δ6 desaturases with novel activities. Also provided are methods and DNA constructs for transiently and/or stably transforming cells, particularly plant cells, with multiple genes.

Claims (21)

1. A process for producing seed comprising eicosapentaenoic acid (EPA) and docosapentaenoic acid (DPA), the process comprising the steps of

(i) growing a Brassicaceae plant comprising exogenous polynucleotides encoding a microalgal Δ6-desaturase which has greater Δ6-desaturase activity on α-linolenic acid (ALA) joined to CoA (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, and a microalgal Δ5-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 a total fatty acid content comprising ALA, stearidonic acid (SDA), eicosatetraenoic acid (ETA), EPA and DPA, wherein the ALA, SDA, ETA, EPA and DPA are each present at a level in the total fatty acid content of the seed, each level being expressed as a percentage of the total fatty acid content, such that the sum of the percentages for EPA and DPA divided by the sum of the percentages for ALA, SDA, ETA, EPA and DPA, expressed as a percentage, is at least 17.3%, and

(ii) harvesting seed from the plant, thereby producing the seed comprising EPA and DPA.

2. The process of claim 1 , wherein arachidonic acid (ARA), EPA, DPA and docosahexaenoic acid (DHA), if present in the total fatty acid content, together comprise 15% of the total fatty acid content in the seed.

3. The process of claim 1 , wherein the Δ6-desaturase is an Ostreococcus tauri Δ6-desaturase, Mantoniella squamata Δ 6-desaturase or Micromonas pusilla Δ 6-desaturase.

4. The process of claim 1 , wherein the Δ6-elongase is a Physcomitrella patens Δ6-elongase or a Pyramimonas CS-0140 Δ6-elongase.

5. The process of claim 1 , wherein the Δ5-desaturase is a Thraustochytrium sp Δ5-desaturase.

6. The process of claim 1 , wherein the Δ6-desaturase is an Ostreococcus tauri, Mantoniella or Micromonas squamata pusilla Δ6-desaturase, the Δ6-elongase is a Physcomitrella patens Δ6-elongase, and the Δ5-desaturase is a Thraustochytrium sp Δ5-desaturase.

7. The process of claim 1 , wherein the seed further comprises an exogenous polynucleotide encoding a Δ15-desaturase.

8. The process of claim 7 , wherein the Δ15-desaturase is a Perilla frutescens Δ15-desaturase.

9. The process of claim 1 , wherein the Brassicaceae plant is Camelina sativa.

10. The process of claim 1 , further comprising a step of extracting oil from the harvested seed.

11. The process of claim 6 , wherein the Brassicaceae plant is Camelina sativa.

12. The process of claim 7 , wherein the Brassicaceae plant is Camelina sativa.

13. The process of claim 8 , wherein the Brassicaceae plant is Camelina sativa.

14. The process of claim 6 , wherein the seed further comprises an exogenous polynucleotide encoding a Δ15-desaturase.

15. The process of claim 14 , wherein the Brassicaceae plant is Camelina sativa.

16. The process of claim 15 , wherein the Δ15-desaturase is a Perilla frutescens Δ15-desaturase.

17. A process for producing seed comprising eicosapentaenoic acid (EPA) and docosapentaenoic acid (DPA), the process comprising the steps of:

(i) growing a Camelina sativa plant comprising exogenous polynucleotides encoding an Ostreococcus tauri Δ 6-desaturase, a Δ6-elongase, and a microalgal Δ5-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 a total fatty acid content comprising ALA, stearidonic acid (SDA), eicosatetraenoic acid (ETA), EPA and DPA, wherein the ALA, SDA, ETA, EPA and DPA are each present at a level in the total fatty acid content of the seed, each level being expressed as a percentage of the total fatty acid content, such that the sum of the percentages for EPA and DPA divided by the sum of the percentages for ALA, SDA, ETA, EPA and DPA, expressed as a percentage, is at least 17.3%, and

(ii) harvesting seed from the plant, thereby producing the seed comprising EPA and DPA.

Continuity (7)
Continuation 16860936 · Apr 28, 2020
Continuation 15642223 · Jul 5, 2017
Continuation 14332202 · Jul 15, 2014
Continuation 13129940
Provisional Application 61270710 · Jul 9, 2009
Provisional Application 61119669 · Nov 18, 2008
Related Publication 20240150779A1 · May 9, 2024
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