Enzymes and methods for producing omega-3 fatty acids

• US 8,809,559 B2
• Filed: 11/17/2009
• Issued: 08/19/2014
• Est. Priority Date: 11/18/2008
• 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), Grains Research & Development Corporation
• 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: Petrie, James Robertson, Liu, Qing, Shrestha, Pushkar, Nichols, Peter David, Blackburn, Susan Irene Ellis, Robert, Stanley Suresh, Zhou, Xue-Rong, Singh, Surinder Pal, Mackenzie, Anne Maree, Mansour, Maged Peter, Frampton, Dion Matthew Frederick, Wood, Craig Christopher
• Primary Examiner(s): Cutliff, Yate K
• Application Number:
  US13/129,940
• Publication Number:
  US 20120016144A1
• Time in Patent Office: 1,736 Days
• Field of Search: None
• US Class Current: 554/9
• 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 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• 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 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/24 Antidepressants
• 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 25/32 Alcohol-abuse
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• 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 3/04 : Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/24 : of the sex hormones
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/06 : Antiarrhythmics
• 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: C12N 15/52 : Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/8202 : by biological means, e.g. cell mediated or natural vector
• CPC Class Codes: C12N 15/8205 : Agrobacterium mediated transformation
• CPC Class Codes: C12N 15/8216 : Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8218 : Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 15/8247 : involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/8257 : for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 9/0071 : acting on paired donors with incorporation of molecular oxygen (1.14)
• 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: C12N 9/93 : Ligases (6)
• 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 7/6463 : obtained from glyceride producing microorganisms, e.g. single cell oil
• 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: C12Y 114/00 : Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
• CPC Class Codes: C12Y 114/19 : with oxidation of a pair of donors resulting in the reduction of molecular oxygen to two molecules of water (1.14.19)
• CPC Class Codes: C12Y 114/19003 : Linoleoyl-CoA desaturase (1.14.19.3)
• CPC Class Codes: C12Y 602/01003 : Long-chain-fatty-acid-CoA ligase (6.2.1.3)

Abstract

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.

Claims

1. A process for producing oil comprising eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, the process comprising the steps of (i) obtaining seed from a canola plant, the seed comprising eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid which were produced from α

-linolenic acid 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) extracting oil from the seed so as to thereby produce the oil.

2. The process of claim 1, wherein arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid together comprise at least 15% of the total fatty acid in the oil.

3. The process of claim 2, wherein the total fatty acid in the oil comprises at least 4% docosahexaenoic acid.

4. The process of claim 3, wherein at least 6% of the total fatty acid incorporated in triacylglycerol in the oil is docosahexaenoic acid.

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

6. The process of claim 5 further comprising a step of degumming, caustic refining, bleaching or deodorizing the extracted oil.

7. The process of claim 1, wherein docosapentaenoic acid and docosahexaenoic acid are present at levels based on en efficiency of conversion of α

-linolenic acid to docosahexaenoic acid of at least 15.4%.

8. 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%.

9. 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%.