Production of High Levels of DHA-Containing Biomass in Microalgae Using Modified Amounts of Chloride and Potassium

Patent metadata

Field	Value
Publication number	US-2016108443-A1
Application number	US-201514975670-A
Country	US
Kind code	A1
Filing date	Dec 18, 2015
Priority date	Oct 2, 2003
Publication date	Apr 21, 2016
Grant date	—

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

1 .- 21 . (canceled)

22 . A method of producing a biomass comprising docosahexaenoic acid (DHA), 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; potassium ions at a concentration of 1.5 g/L to 10 g/L; and recovering DHA-containing biomass from the microalga.

23 . The method of claim 22 , wherein the concentration of chloride ions is less than or equal to 2 g/L.

24 . The method of claim 22 , wherein the concentration of chloride ions is less than or equal to 1 g/L.

25 . The method of claim 22 , wherein the concentration of chloride ions is less than or equal to 0.3 g/L.

26 . The method of claim 22 , wherein the concentration of potassium ions is 1.5 g/L to 4 g/L.

27 . The method of claim 22 , wherein the concentration of potassium ions is 1.5 g/L to 3 g/L.

28 . The method of claim 22 , wherein the concentration of potassium ions is 1.5 g/L to 2.4 g/L.

29 . The method of claim 22 , wherein the concentration of potassium ions is 1.5 g/L to 2 g/L.

30 . The method of claim 22 , wherein the culture medium further comprises sodium ions at a concentration of 1 g/L to 8 g/L.

31 . The method of claim 22 , 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.

32 . The method of claim 22 , 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.

33 . The method of claim 22 , wherein a source of potassium comprises potassium sulfate.

34 . The method of claim 22 , wherein the microalga produces at least 0.10 g DHA per 10 9 cells.

35 . The method of claim 22 , wherein the microalga produces at least 0.20 g DHA per 10 9 cells.

36 . The method of claim 22 , wherein the culture medium further comprises sodium ions and the ratio of sodium ions to potassium ions is less than or equal to 15:1.

37 . The method of claim 22 , 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.

38 . The method of claim 22 , further comprising recovering DHA from the DHA-containing lipid.

39 . The method of claim 22 , wherein the microalga is of the genus Dinophyceae.

40 . The method of claim 22 , wherein the microalga is of the genus Crypthecodinium.

41 . The method of claim 22 , wherein the microalga is of the species Crypthecodinium cohnii.

42 . The method of claim 22 , wherein the culture medium further comprises sodium ions, wherein a ratio of sodium ions to potassium ions is less than or equal to 27:1.

43 . The method of claim 22 , wherein the microalga produces at least 0.04 g DHA per 10 9 cells.

44 . A lipid extracted from the biomass of claim 22 .

45 . A method of producing docosahexaenoic acid (DHA), comprising:
culturing a heterotrophic microalga in a culture medium to produce said DHA, wherein the culture medium comprises:
chloride ions at a concentration of less than or equal to 2 g/L;
potassium ions at a concentration of greater than or equal to 0.5 g/L; and
the microalga produces at least 51% DHA as a weight percent of fat; and
recovering the DHA from the microalga.

Assignees

• Dsm Ip Assets Bv

Inventors

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

Classifications

• C12N1/12

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

• A61K36/02

  Algae · CPC title

• 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

• C12P7/6409

  Fatty acids · CPC title