Eukaryotic microorganisms for producing lipids and antioxidants

• US 10,435,725 B2
• Filed: 06/21/2017
• Issued: 10/08/2019
• Est. Priority Date: 06/07/2005
• Status: Active Grant

• Current Assignee: DSM Nutritional Products AG (DSM-Firmenich AG)
• Original Assignee: DSM Nutritional Products AG (DSM-Firmenich AG)
• Inventors: Burja, Adam M., Radianingtyas, Helia, Barrow, Colin James, Windust, Anthony James
• Primary Examiner(s): Shen, Bin
• Application Number:
  US15/629,510
• Publication Number:
  US 20170335356A1
• Time in Patent Office: 839 Days
• CPC Class Codes: A21D 8/04 treating dough with microorganisms or enzymes
• CPC Class Codes: A23K 10/18 of live microorganisms
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23L 29/065 Microorganisms addition of bacteria for nutritional purposes A23L33/135
• 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: A23P 10/30 Encapsulation of particles, e.g. foodstuff additives
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A23V 2200/02 Antioxidant
• CPC Class Codes: A23V 2200/3262 having an effect on blood cholesterol
• CPC Class Codes: A23V 2200/328 having effect on glycaemic control and diabetes
• CPC Class Codes: A23V 2200/332 Promoters of weight control and weight loss
• 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 9/4875 Compounds of unknown constitution, e.g. material from plants or animals oils, fats, waxes, shellac A61K9/4858
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 39/06 : Free radical scavengers or antioxidants
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 1/00 : Microorganisms, e.g. protozoa; Compositions thereof medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
• 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: C12R 2001/00 : Microorganisms ; Processes using microorganisms

Abstract

Disclosed are compositions and methods related to eukaryotic microorganisms that can produce unsaturated fatty acids which can be purified and used.

Claims

1. A method of increasing biomass production of an isolated eukaryotic microorganism the method comprising:

(a) culturing in a heterotrophic medium an isolated eukaryotic microorganism having an 18S sequence, wherein the 18S sequence has at least 97% identity to the sequence set forth in SEQ ID NO;

1 and

(b) increasing biomass production by increasing agitation of the culture, increasing the concentration of dO₂ in the culture, and increasing the temperature of the culture or a combination thereof.

2. The method of claim 1, wherein the 18S sequence has at least 99% identity to the sequence set forth in SEQ ID NO:

1.

3. The method of claim 1, wherein the microorganism has ATCC Accession Number PTA-6245.

4. The method of claim 1, wherein the microorganism is a Schizochytrium sp.

5. The method of claim 1, wherein the microorganism is a Thraustochytrium sp.

6. The method of claim 1, wherein the heterotrophic medium comprises a carbon source in an amount of from 1 to 60 g L⁻

1.

7. The method of claim 1, wherein the heterotrophic medium comprises a carbon source in an amount of from 1 to 200 g L⁻

1.

8. The method of claim 1, wherein the method further comprises isolating the biomass.

9. The method of claim 1, wherein the method further comprises harvesting the eukaryotic microorganism.

10. The method of claim 9, wherein the method further comprises rupturing or permeabilizing the harvested microorganism.

11. The method of claim 10, wherein the rupturing or permeabilizing comprises sonication, liquid-shear disruption methods, bead milling, pressing under high pressure, freeze-thawing, or enzymatic digestion.

12. The method of claim 10, wherein the rupturing or permeabilizing comprises enzymatic digestion.

13. The method of claim 1, wherein the biomass comprises lipids and the method further comprises isolating the lipids from the biomass.

14. The method of claim 13, wherein the biomass comprises neutral lipids.

15. The method of claim 14, wherein the neutral lipids comprise at least 95% by weight of total lipids.

16. The method of claim 13, wherein the lipids are isolated using a solvent.

17. The method of claim 16, wherein the solvent is selected from the group consisting of chloroform, hexane, methanol, ethanol, and supercritical CO₂.

18. The method of claim 16, wherein the method further comprises removing the solvent.