Synthesis of long-chain polyunsaturated fatty acids by recombinant cells

• US 7,807,849 B2
• Filed: 04/22/2005
• Issued: 10/05/2010
• Est. Priority Date: 04/22/2004
• Status: Active Grant

• Current Assignee: Commonwealth Scientific & Industrial Research Organisation (Commonwealth Of Australia As Represented By Department Of Industry, Innovation, Climate Change, Science, Research And Tertiary Education)
• Original Assignee: Commonwealth Scientific & Industrial Research Organisation (Commonwealth Of Australia As Represented By Department Of Industry, Innovation, Climate Change, Science, Research And Tertiary Education)
• Inventors: Zhou, Xue-Rong, Singh, Surinder Pal, Blackburn, Susan Irene Ellis, Robert, Stanley Suresh, Petrie, James Robertson, Nichols, Peter David, Green, Allan Graham
• Primary Examiner(s): Carr; Deborah D
• Application Number:
  US11/112,882
• Publication Number:
  US 20050273885A1
• Time in Patent Office: 1,992 Days
• Field of Search: 554/227
• US Class Current: 554/227
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/04 for urolithiasis
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: A61P 9/14 Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C07C 51/42 Separation; Purification; Stabilisation; Use of additives
• CPC Class Codes: C11B 1/025 : by enzymes or microorganisms, living or dead
• CPC Class Codes: C11B 1/06 : by pressing
• CPC Class Codes: C11B 3/006 : by extraction
• CPC Class Codes: C12N 15/8247 : involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0083 : Miscellaneous (1.14.99)
• CPC Class Codes: C12N 9/1029 : transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6427 : Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6432 : Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 : Docosahexenoic acids [DHA]
• CPC Class Codes: C12P 7/6472 : containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
• CPC Class Codes: Y02P 20/52 : using catalysts, e.g. selective catalysts

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 poly unsaturated fatty acids.

Claims

1. A process for producing oil containing eicosapentaenoic acid, docosahexaenoic acid and docosapentaenoic acid, comprising the steps of obtaining a tranagenic oil-seed rape seed, a tranagenic cotton seed or a tranagenic flax seed comprising eicosapentaenoic acid, docosahexaenoic acid and docosapentaenoic acid, wherein at least 25% (w/w) of the eiscosapentaenoic acid, docosahexaenoic acid and docosapentaenoic acid of the transgenic seed is incorporated into triacylglycerols in the transgenic seed, and wherein the total fatty acid in the oil of the transgenic seed comprises at least 2.5% ω

3 C20 fatty acids (w/w), and extracting oil from the transgenic seed so as to thereby produce the oil.

2. The process of claim 1, wherein the step of extracting the oil comprises crushing the transgenic seed.

3. The process of claim 1, wherein the total fatty acid in the oil of the transgenic seed comprises at least 9% C20 fatty acids (w/w)

4. The process of claim 1, wherein the total fatty acid in the oil of the tranagenic seed comprises at least 1.5% eicosapentaenoic acid and at least 0.13% docosapentaenoic acid (w/w).

5. The process of claim 1, wherein the level of docosapentaenoic acid relative to eicosapentaenoic acid in the transgenic seed is at least 5% (w/w).

6. The process of claim 1, wherein the total fatty acid in the oil of the tranagenic seed comprises at least 2.1% eicosapentaenoic acid and less than 0.1% eicosatrienoic acid (w/w).

7. The process of claim 1, wherein the tranagenic seed is a cotton seed.

8. The process of claim 1, wherein the transgenic seed is a flax seed.

9. The process of claim 1, wherein the transgenic seed is an oilseed rape seed.

10. The process of claim 1, wherein at least 50% (w/w) of the eicosapentasnoic acid, docosahexaenoic acid and docosapentaenoic acid of the transgenic seed is incorporated into triacylglycerols in the seed.

11. The process of claim 1, wherein the oil comprises at least 50% triacylglycerols.