IP Library Granted Patent US 9,848,623
Granted Patent B2
US 9,848,623 · App. 13/342,623 · Granted Dec 26, 2017

Enhanced production of lipids containing polyenoic fatty acids by very high density cultures of eukaryotic microbes in fermentors

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Quick Facts
Patent No.
US 9,848,623
App. No.
13/342,623
Granted
Dec 26, 2017
Kind
B2
Abstract

The present invention provides a process for growing eukaryotic microorganisms which are capable of producing lipids, in particular lipids containing polyenoic fatty acids. The present invention also provides a process for producing eukaryotic microbial lipids.

Claims (34)

1. A process for producing microbial lipids comprising conducting a fermentation of microorganisms of the order Thraustochytriales capable of producing at least 15% of lipids as highly unsaturated fatty acids having at least four unsaturated carbon-carbon bonds in a fermentation medium comprising a carbon source and a limiting nutrient source, the process comprising:

a) providing the fermentation medium comprising the carbon source and limiting nutrient source;

b) adding the microorganisms to the fermentation medium;

c) adding an additional amount of the carbon source and limiting nutrient source to a fermentation medium to increase the biomass density of the fermentation medium to at least 100 g/L on a dry cell weight basis; and

d) adding an additional amount of the carbon source while limiting the amount of the limiting nutrient source added to the formulation medium to induce lipid production by the microorganisms.

2. The process of claim 1 , further comprising e) recovering at least some of the lipids from the microorganisms.

3. The process of claim 1 , wherein the fermentation of the microorganisms produces lipids containing highly unsaturated fatty acids when the biomass density of the fermentation medium is at least 150 g/L on a dry cell weight basis.

4. The process of claim 1 , wherein the process produces lipids at an average rate of at least 0.5 grams lipids per liter of fermentation medium per hour fermentation.

5. The process of claim 1 , wherein the microorganisms are selected from the group consisting of Thraustochytrium, Schizochytrium , and mixtures thereof.

6. The process of claim 1 , wherein the process produces on average at least 0.2 grams docosahexaenoic acid per liter of fermentation medium per hour fermentation.

7. The process of claim 1 , wherein the microorganisms produce lipids containing highly unsaturated fatty acids when the biomass density is at least 100 g/L on a dry cell weight basis.

8. The process of claim 1 , wherein the highly unsaturated fatty acids are selected from the group consisting of docosahexaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, arachidonic acid and mixtures thereof.

9. The process of claim 8 , wherein the highly unsaturated fatty acids are docosahexaenoic acid.

10. The process of claim 1 , wherein at least 25% of the lipids is docosahexaenoic acid.

11. The process of claim 1 , wherein the process produces lipids at an average rate of at least 0.5 g/L/hr and wherein at least 25% of the lipids is docosahexaenoic acid.

12. The process of claim 1 , wherein the carbon source comprises a carbohydrate.

13. The process of claim 1 , wherein the limiting nutrient source comprises a nitrogen source.

14. The process of claim 1 , wherein the pH of the fermentation medium is about pH 5 to about pH 11.

15. The process of claim 1 , wherein the fermentation medium is at a temperature of at least 20° C.

16. The process of claim 1 , wherein the microorganisms produce highly unsaturated fatty acids under aerobic conditions.

17. The process of claim 1 , wherein dissolved oxygen in the fermentation medium is controlled.

18. The process of claim 1 , further comprising: removing water from the fermentation medium to provide dry microorganisms prior to recovering the lipids from the microorganisms, wherein at least 15% of the lipids are highly unsaturated fatty acids.

19. The process of claim 18 , wherein the water is removed from the fermentation medium by evaporation without prior centrifugation to provide dry microorganisms prior to recovering the lipids from the microorganisms, wherein at least 15% of the microbial lipids are highly unsaturated fatty acids.

20. A process for growing microorganisms of the order Thraustochytriales by conducting a fermentation of the microorganisms in a fermentation medium comprising a carbon source, and a limiting nutrient source the process comprising:

a) providing the fermentation medium comprising the carbon source and the limiting nutrient source;

b) adding the microorganisms to the fermentation medium;

c) adding an additional amount of the carbon source and limiting nutrient source to the fermentation medium to increase the biomass density of the fermentation medium to at least 100 g/L on a dry cell weight basis, wherein the additional amount of the carbon source and the limiting nutrient source are added at a rate sufficient to increase the biomass density of the fermentation medium to at least 100 g/L;

d) adding an additional amount of the carbon source while reducing the amount of limiting nutrient source added to induce lipid production by the microorganisms; and

e) recovering the lipids from the fermentation medium,

wherein the primary use of the carbon source by the microorganisms is to produce the lipids after induction of lipid production;

wherein the fermentation of the microorganisms when the biomass density is at least 100 g/L on a dry cell weight basis produces lipids containing polyunsaturated fatty acids;

wherein the lipids are at least 20% of the biomass when the biomass density is at least 100 g/L;

wherein the process produces lipids at least at an average rate of 0.5 g/L/hr; and

wherein at least about 15% of the total lipids produced by the microorganisms are polyunsaturated lipids.

Assignments (5)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 11, 2012
From: MARTEK BIOSCIENCES CORPORATION
To: DSM IP ASSETS B.V.
Reel/Frame 028938/0620 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 8, 2012
From: MARTEK BIOSCIENCES BOULDER CORPORATION
To: MARTEK BIOSCIENCES CORPORATION
Reel/Frame 028921/0883 →
MERGER Recorded Sep 8, 2012
From: OMEGATECH, INC.
To: OMEGATECH, INC.
Reel/Frame 028921/0917 →
CHANGE OF NAME Recorded Sep 8, 2012
From: OMEGATECH, INC.
To: MARTEK BIOSCIENCES BOULDER CORPORATION
Reel/Frame 028923/0383 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 7, 2012
From: BAILEY, RICHARD B.; DIMASI, DON; HANSEN, JON M.; MIRRASOUL, PETER J.; RUECKER, CRAIG M.; VEEDER, GEORGE T., III; KANEKO, TATSUO; BARCLAY, WILLIAM R.
To: OMEGATECH, INC.
Reel/Frame 028919/0747 →