Thraustochytrids, Fatty Acid Compositions, and Methods of Making and Uses Thereof

• US 20100239533A1
• Filed: 03/19/2009
• Published: 09/23/2010
• Est. Priority Date: 03/19/2009
• Status: Active Grant

• Current Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Original Assignee: Martek Biosciences Corporation (DSM-Firmenich AG)
• Inventors: Hansen, Jon Milton, APT, Kirk E., Zirkle, Ross, Pfeifer, Joseph W. III, Behrens, Paul Warren, Stahl, Tracey Lynn
• Granted Patent: US 8,207,363 B2
• Time in Patent Office: Days
• US Class Current: 424/93.100
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23K 50/00 Feeding-stuffs specially adapted for particular animals
• CPC Class Codes: A23K 50/80 for aquatic animals, e.g. fish, crustaceans or molluscs
• CPC Class Codes: A23L 33/115 Fatty acids or derivatives thereof; Fats or oils
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A23L 33/135 Bacteria or derivatives thereof, e.g. probiotics
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A61K 2035/11 Medicinal preparations comprising living procariotic cells
• CPC Class Codes: A61K 2800/74 Biological properties of particular ingredients
• CPC Class Codes: A61K 31/20 having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
• CPC Class Codes: A61K 31/202 having three or more double bonds, e.g. linolenic eicosanoids, e.g. leukotrienes A61K31/557
• CPC Class Codes: A61K 36/02 Algae
• CPC Class Codes: A61K 8/9728 Fungi, e.g. yeasts
• CPC Class Codes: A61K 8/99 from microorganisms other than algae or fungi, e.g. protozoa or bacteria
• 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: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: C12N 1/125 Unicellular algae isolates
• 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: C12R 2001/89 : Algae ; Processes using algae

Abstract

The present invention is directed to isolated thraustochytrid microorganisms as well as strains and mutants thereof. The invention is further directed to biomasses, microbial oils, compositions, cultures, methods of producing microbial oils, and methods of using the isolated thraustochytrids, biomasses, and microbial oils.

Claims

1. An isolated thraustochytrid microorganism of the thraustochytrid species deposited under ATCC Accession No. PTA-9695 or a strain derived therefrom, wherein the total fatty acids produced by said microorganism or strain derived therefrom comprise about 10% or less by weight eicosapentaenoic acid.

5. The isolated thraustochytrid microorganism of any one of claims 1 to 4, wherein the strain derived therefrom is a mutant strain.

10. A thraustochytrid biomass comprising the thraustochytrid microorganism of any one of claims 1 to 9 or mixtures thereof.

14. The isolated thraustochytrid biomass of any one of claims 10 to 13, wherein about 10% or less by weight of the fatty acids is eicosapentaenoic acid, and wherein the weight ratio of docosahexaenoic acid to eicosapentaenoic acid is at least about 5:

1.

15. The isolated thraustochytrid biomass of any one of claims 10 to 14, wherein about 1.5% or less by weight of the fatty acids is arachidonic acid, and wherein the weight ratio of docosahexaenoic acid to arachidonic acid is at least about 20:

1.

16. The isolated thraustochytrid biomass of any one of claims 10 to 15, further comprising docosahexaenoic acid and docosapentaenoic acid n-6 in a weight ratio of at least about 10:

1.

17. An isolated thraustochytrid culture comprising the thraustochytrid microorganism of any one of claims 1 to 9 or mixtures thereof.

18. The isolated thraustochytrid culture of claim 17, further comprising at least about 5% dissolved oxygen.

19. A food product, cosmetic, or pharmaceutical composition for animals or humans, comprising the thraustochytrid microorganism or thraustochytrid biomass of any one of claims 1 to 16 or mixtures thereof.

28. The food product of claim 19 or claim 27, wherein the food product is an infant formula.

29. The food product of claim 28, wherein the infant formula is suitable for premature infants.

30. The food product of claim 19 or claim 27, wherein the food product is milk, a beverage, a therapeutic drink, a nutritional drink, or a combination thereof.

31. The food product of claim 19 or claim 27, wherein the food product is an additive for animal or human food.

32. The food product of claim 19 or claim 27, wherein the food product is a nutritional supplement.

33. The food product of claim 19 or claim 27, wherein the food product is an animal feed.

34. The animal feed of claim 33, wherein the animal feed is an aquaculture feed.

35. The animal feed of claim 33, wherein the animal feed is a domestic animal feed, a zoological animal feed, a work animal feed, a livestock feed, or a combination thereof.

36. A method for producing a microbial oil comprising omega-3 fatty acids, the method comprising:

(a) growing the isolated thraustochytrid microorganism of any one of claims 1 to 9 or mixtures thereof in a culture to produce a biomass, and

(b) extracting an oil comprising omega-3 fatty acids from the biomass.

37. The method of claim 36, wherein the culture comprises at least about 5% dissolved oxygen.

38. The method of claim 36, wherein the culture pH is maintained at about 6.5 to about 8.5.

39. The method of claim 36, wherein the culture temperature is maintained at about 17°

C. to about 30°

C.

40. The method of claim 36, wherein the culture comprises a glucose concentration of about 5 g/L to about 50 g/L.

41. A method for producing a microbial oil comprising omega-3 fatty acids, the method comprising extracting an oil comprising omega-3 fatty acids from the biomass of any one of claims 10 to 16.

42. The method of claim 41, wherein the microbial oil comprising omega-3 fatty acids is extracted using a hexane extraction process.

43. The method of claim 41, wherein the microbial oil comprising omega-3 fatty acids is extracted using a solventless extraction process.

44. The method of any one of claims 36 to 43, or the isolated thraustochytrid culture of claim 17 or claim 18, wherein the dry cell weight of biomass isolated from each liter of culture is at least about 50 g after growing for about 7 days at about 17°

C. to about 30°

C. in a culture medium of about pH 6.5 to about pH 8.0 comprising sources of carbon, nitrogen, and nutrients, and about 950 ppm to about 8500 ppm chloride ions.

45. The method of any one of claims 36 to 43, or the isolated thraustochytrid culture of claim 17 or claim 18, wherein the omega-3 fatty acids productivity is at least about 2 g/L/day after growing for about 7 days at about 17°

C. to about 30°

C. in a culture medium of about pH 6.5 to about pH 8.0 comprising sources of carbon, nitrogen, and nutrients, and about 950 ppm to about 8500 ppm chloride ions.

46. A microbial oil produced by the method of any one of claims 36 to 45.

47. Use of the isolated microorganism, biomass, or microbial oil of any one of claims 1 to 16, 20 to 26, 46, or mixtures thereof, for the manufacture of a medicament for the treatment of inflammation or a condition related thereto.

48. Use of the isolated microorganism, biomass, or microbial oil of any one of claims 1 to 16, 20 to 26, 46, or mixtures thereof, for the treatment of inflammation or a condition related thereto.

49. The isolated microorganism, biomass, or microbial oil of any one of claims 1 to 16, 20 to 26, 46, or mixtures thereof, for use in the treatment of inflammation or a condition related thereto.

50. A method for treating inflammation or a condition related thereto in a subject in need thereof, comprising administering to the subject the isolated microorganism, biomass, or microbial oil of any one of claims 1 to 16, 20 to 26, 46, or mixtures thereof, and a pharmaceutically acceptable carrier.

2. An isolated thraustochytrid microorganism having the characteristics of the thraustochytrid species deposited under ATCC Accession No. PTA-9695, wherein the total fatty acids produced by said microorganism or strain derived therefrom comprise about 10% or less by weight eicosapentaenoic acid.

3. An isolated thraustochytrid microorganism, or a strain derived therefrom, comprising a triglyceride fraction, wherein the docosahexaenoic acid content of the triglyceride fraction is at least about 40% by weight, wherein the docosapentaenoic acid n-6 content of the triglyceride fraction is from at least about 0.5% by weight to about 6% by weight, and wherein the total fatty acids produced by said microorganism or strain derived therefrom comprise about 10% or less by weight of eicosapentaenoic acid.

4. An isolated thraustochytrid microorganism of the same species as the thraustochytrid deposited under ATCC Accession No. PTA-9695, or a strain derived therefrom, wherein the total fatty acids produced by said microorganism or strain derived therefrom comprise about 10% or less by weight eicosapentaenoic acid.

6. An isolated thraustochytrid microorganism deposited under ATCC Accession No. PTA-9695.

7. An isolated thraustochytrid microorganism deposited under ATCC Accession No. PTA-9696.

8. An isolated thraustochytrid microorganism deposited under ATCC Accession No. PTA-9697.

9. An isolated thraustochytrid microorganism deposited under ATCC Accession No. PTA-9698.

11. An isolated thraustochytrid biomass, wherein at least about 50% by weight of the dry cell weight of the biomass are fatty acids, and wherein at least about 50% by weight of the fatty acids are omega-3 fatty acids.

12. The isolated thraustochytrid biomass of claim 11, wherein at least about 50% by weight of the fatty acids is docosahexaenoic acid.

13. An isolated thraustochytrid biomass, wherein at least about 25% by weight of the dry cell weight of the biomass is docosahexaenoic acid.

20. A microbial oil comprising a triglyceride fraction of at least about 70% by weight, wherein the docosahexaenoic acid content of the triglyceride fraction is at least about 50% by weight, and wherein the docosapentaenoic acid n-6 content of the triglyceride fraction is from about 0.5% by weight to about 6% by weight.

21. The microbial oil of claim 20, further comprising an arachidonic acid content of the triglyceride fraction of about 1.5% or less by weight.

24. The microbial oil of any one of claims 20 to 23, wherein at least about 20% of the triglycerides in the triglyceride fraction contain docosahexaenoic acid at two positions in the triglyceride selected from any two of the sn-1, sn-2, and sn-3 positions.

25. The microbial oil of any one of claims 20 to 23, wherein at least about 5% of the triglycerides in the triglyceride fraction contain docosahexaenoic acid at all three of the sn-1, sn-2, and sn-3 positions in the triglyceride.

26. The microbial oil of any one of claims 20 to 23, further comprising about 5% by weight or less of heptadecanoic acid.

27. A food product, cosmetic, or pharmaceutical composition for animals or humans, comprising the microbial oil of any one of claims 20 to 26.

22. A microbial oil comprising a triglyceride fraction of at least about 70% by weight, wherein the docosahexaenoic acid content of the triglyceride fraction is at least about 40% by weight, wherein the docosapentaenoic acid n-6 content of the triglyceride fraction is from at least about 0.5% by weight to about 6% by weight, and wherein the ratio of docosahexaenoic acid to docosapentaenoic acid n-6 is greater than about 6:

1.

23. A microbial oil comprising a triglyceride fraction of at least about 70% by weight, wherein the docosahexaenoic acid content of the triglyceride fraction is at least about 60% by weight.