Eukaryotic microorganisms for producing lipids and antioxidants
Disclosed are compositions and methods related to eukaryotic microorganisms that can produce unsaturated fatty acids which can be purified and used.
1. A method of increasing biomass production of an isolated eukaryotic microorganism, the method comprising:
(i) 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
(ii) increasing agitation of the culture to 100 to 1000 rpm, increasing the concentration of dO2 in the culture to between 1 and 10%, increasing the temperature of the culture to 4 to 30° C., or a combination thereof, wherein biomass production is increased.
2. The method of claim 1 , further comprising isolating the biomass.
3. The method of claim 2 , wherein isolating the biomass comprises harvesting the eukaryotic microorganism.
4. The method of claim 1 , wherein the isolated eukaryotic microorganism has ATCC accession number PTA-6245.
5. The method of claim 1 , wherein culturing decreases docosahexaenoic acid (DHA) production.
6. The method of claim 1 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 60 g L −1 .
7. The method of claim 1 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 200 g L −1 .
8. The method of claim 7 , wherein the harvested eukaryotic microorganism is ruptured or permeabilized.
9. The method of claim 7 , wherein the harvested eukaryotic microorganism is ruptured or permeabilized by sonication, liquid-shear disruption methods, bead milling, pressing under high pressure, freeze-thawing, or enzymatic digestion of the cell wall.
10. A method of increasing fatty acid production of an isolated eukaryotic microorganism, the method comprising:
(i) 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
(ii) increasing the agitation of the culture to 100 to 1000 rpm, increasing the concentration of dO2 in the culture to between 1 and 10%, increasing the temperature of the culture to 4 to 30° C., or a combination thereof, wherein fatty acid production is increased.
11. The method of claim 10 , further comprising isolating the fatty acids.
12. The method of claim 10 , wherein the isolated eukaryotic microorganism has ATCC accession number PTA-6245.
13. The method of claim 10 , wherein culturing decreases DHA production.
14. The method of claim 10 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 60 g L −1 .
15. The method of claim 10 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 200 g L −1 .
16. A method of increasing lipid production of an isolated eukaryotic microorganism, the method comprising:
(i) 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
(ii) increasing the agitation of the culture to 100 to 1000 rpm, increasing the concentration of dO2 in the culture to between 1 and 10%, increasing the temperature of the culture to 4 to 30° C., or a combination thereof, wherein lipid production is increased.
17. The method of claim 16 , further comprising isolating the lipid.
18. The method of claim 16 , wherein the isolated eukaryotic microorganism has ATCC accession number PTA-6245.
19. The method of claim 16 , wherein culturing decreases DHA production.
20. The method of claim 16 , wherein the lipid is isolated with a solvent.
21. The method of claim 20 , wherein the solvent comprises chloroform, hexane, methanol, ethanol, or supercritical CO 2 .
22. The method of claim 21 , further comprising hydrolyzing the lipid, wherein hydrolyzing the lipid composition converts triglycerides to free fatty acids or esters of fatty acids.
23. The method of claim 20 , further comprising removing the solvent.
24. The method of claim 16 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 60 g L −1 .
25. The method of claim 16 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 200 g L −1 .
26. A method of decreasing biomass production of an isolated eukaryotic microorganism, the method comprising:
(i) 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
(ii) decreasing agitation of the culture to 100 to 1000 rpm, decreasing the concentration of dO2 in the culture to between 1 and 10%, decreasing the temperature of the culture to 4 to 30° C., or a combination thereof, wherein biomass production is decreased.
27. The method of claim 26 , further comprising isolating the biomass.
28. The method of claim 27 , wherein isolating the biomass comprises harvesting the eukaryotic microorganism.
29. The method of claim 26 , wherein the isolated eukaryotic microorganism has ATCC accession number PTA-6245.
30. The method of claim 27 , wherein culturing increases DHA production.
31. The method of claim 26 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 60 g L −1 .
32. The method of claim 26 , wherein the heterotrophic medium comprise a carbon source in an amount of from 1 g L −1 to 200 g L −1 .
33. The method of claim 32 , wherein the harvested eukaryotic microorganism is ruptured or permeabilized.
34. The method of claim 32 , wherein the harvested eukaryotic microorganism is ruptured or permeabilized by sonication, liquid-shear disruption methods, bead milling, pressing under high pressure, freeze-thawing, or enzymatic digestion of the cell wall.