Production of high levels of DHA-containing biomass in microalgae using modified amounts of chloride and potassium

Patent metadata

Field	Value
Publication number	US-9834799-B2
Application number	US-201514975670-A
Country	US
Kind code	B2
Filing date	Dec 18, 2015
Priority date	Oct 2, 2003
Publication date	Dec 5, 2017
Grant date	Dec 5, 2017

Abstract

Methods for production of highly unsaturated fatty acids by marine microorganisms, including the heterotrophic marine dinoflagellate Crypthecodinium , using low levels of chloride ion are disclosed. Specifically, methods of increasing production of highly unsaturated fatty acids by marine microorganisms while growing in low chloride media by manipulating sodium ion and potassium ion levels. The invention also relates to methods of production of highly unsaturated fatty acids by marine organisms at low pH levels, and includes methods for generation of low pH tolerant strains.

First claim

We claim:

1. A method of producing a biomass comprising a lipid fraction comprising docosahexaenoic acid (DHA), the method comprising:
culturing a heterotrophic microalga in a culture medium, wherein the culture medium comprises:
chloride ions at a concentration of less than or equal to 3 g/L; and
potassium ions at a concentration of 1.5 g/L to 10 g/L;
wherein the method further comprises recovering the DHA-containing biomass from the microalga.

2. The method of claim 1 , wherein the concentration of chloride ions is less than or equal to 2 g/L.

3. The method of claim 1 , wherein the concentration of chloride ions is less than or equal to 1 g/L.

4. The method of claim 1 , wherein the concentration of chloride ions is less than or equal to 0.3 g/L.

5. The method of claim 1 , wherein the concentration of potassium ions is 1.5 g/L to 4 g/L.

6. The method of claim 1 , wherein the concentration of potassium ions is 1.5 g/L to 3 g/L.

7. The method of claim 1 , wherein the concentration of potassium ions is 1.5 g/L to 2.4 g/L.

8. The method of claim 1 , wherein the concentration of potassium ions is 1.5 g/L to 2 g/L.

9. The method of claim 1 , wherein the culture medium further comprises sodium ions at a concentration of 1 g/L to 8 g/L.

10. The method of claim 1 , wherein the culture medium further comprises sodium ions at a concentration of 1 g/L to 8 g/L, and wherein the pH is less than or equal to 5.

11. The method of claim 1 , wherein the culture medium further comprises sodium ions at a concentration of 1.5 g/L to 5 g/L, and wherein the pH is 4.5.

12. The method of claim 1 , wherein a source of potassium comprises potassium sulfate.

13. The method of claim 1 , wherein the microalga produces at least 0.10 g DHA per 10 9 cells.

14. The method of claim 1 , wherein the microalga produces at least 0.20 g DHA per 10 9 cells.

15. The method of claim 1 , wherein the culture medium further comprises sodium ions and the ratio of sodium ions to potassium ions is less than or equal to 9:1.

16. The method of claim 1 , wherein the culture medium further comprises sodium ions and the ratio of sodium ions to potassium ions is less than or equal to 4:1.

17. The method of claim 1 , further comprising recovering DHA from the DHA-containing lipid fraction.

18. The method of claim 1 , wherein the microalga is of the class Dinophyceae.

19. The method of claim 1 , wherein the microalga is of the genus Crypthecodinium.

20. The method of claim 1 , wherein the microalga is of the species Crypthecodinium cohnii.

21. The method of claim 1 , wherein the microalga produces at least 0.04 g DHA per 10 9 cells.

Assignees

• Dsm Ip Assets Bv

Inventors

• Behrens Paul W
• Thompson John M
• Apt Kirk
• Pfeifer Iii Joseph W
• Wynn James P
• Lippmeier James Casey
• Fichtali Jaouad
• Hansen Jon

Classifications

• C12N2500/00

  Specific components of cell culture medium · CPC title

• C12N5/0018 Primary

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N1/12

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• A61K36/02

  Algae · CPC title

• C11B1/10

  by extracting · CPC title