Microorganisms having increased lipid productivity
The present invention provides mutant microorganism that have higher lipid productivity than the wild type microorganisms from which they are derived while biomass at levels that are within approximately 50% of wild type biomass productivities under nitrogen replete conditions. Particular mutants produce at least twice as much FAME lipid as wild type while producing at least 75% of the biomass produced by wild type cells under nitrogen replete conditions. Also provided are methods of producing lipid using the mutant strains.
1. A mutant algal or heterokont microorganism that produces at least 50% more fatty acid methyl] ester-derivatizable lipids (FAME lipids) than a substantially identical control microorganism of the same species and at least 70% of the amount of biomass produced by the control microorganism when the mutant microorganism and control microorganism are cultured under identical conditions under which the control microorganism is producing biomass, wherein the mutant microorganism has attenuated expression of a gene encoding a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24.
2. A mutant algal or heterokont microorganism according to claim 1 , wherein the mutant microorganism and control microorganism are cultured under identical conditions which are nitrogen replete with respect to the control microorganism.
3. A mutant algal or heterokont microorganism according to claim 1 , wherein the control microorganism is a wild type microorganism of the same species as the mutant.
4. A mutant algal or heterokont microorganism according to claim 1 , wherein the mutant microorganism produces at least 50% more FAME lipids than the control microorganism while accumulating at least 70% the amount of biomass accumulated by the control microorganism over a culture period of at least five days.
5. A mutant algal or heterokont microorganism according to claim 4 , wherein the mutant microorganism produces at least 50% more FAME lipids than the control microorganism while accumulating at least 70% the amount of biomass accumulated by the control microorganism over a culture period of at least ten days.
6. A mutant algal or heterokont microorganism according to claim 4 , wherein the mutant microorganism accumulates at least 80% the amount of biomass accumulated by the control microorganism.
7. A mutant algal or heterokont microorganism according to claim 6 , wherein the mutant microorganism accumulates at least 90% the amount of biomass accumulated by the control microorganism.
8. A mutant algal or heterokont microorganism according to claim 7 , wherein the mutant microorganism accumulates at least 95% the amount of biomass accumulated by the control microorganism.
9. A mutant algal or heterokont microorganism according to claim 4 , wherein the mutant microorganism produces at least 75% more FAME lipids than a wild type microorganism of the same species as the mutant.
10. A mutant algal or heterokont microorganism according to claim 9 , wherein the mutant microorganism produces at least 100% more FAME lipids than the control microorganism.
11. A mutant algal or heterokont microorganism according to claim 1 , wherein the mutant microorganism exhibits a fatty acid methyl ester (FAME)/total organic carbon (TOC) ratio at least 30% higher than the FAME/TOC ratio of the control microorganism.
12. A mutant heterokont microorganism according to claim 1 , wherein the heterokont microorganism is a Labyrinthulomycetes selected from the group consisting of the genera: Labryinthula, Labryinthuloides, Thraustochytrium, Schizochytrium, Aplanochytrium, Aurantiochytrium, Oblongichytrium, Japonochytrium, Diplophrys , or Ulkenia.
13. A mutant heterokont microorganism according to claim 1 , wherein the heterokont microorganism is a Labyrinthulomycetes species selected from a group consisting of the genera Thraustochytrium, Schizochytrium, Aurantiochytrium, Oblongichytrium , and Japonochytrium.
14. A mutant algal microorganism according to claim 1 , wherein the algal microorganism is a Eustigmatophyte.
15. A mutant algal microorganism according to claim 14 , wherein the Eustigmatophyte is of the genus Nannochloropsis.
16. A method of producing lipid, comprising culturing the microorganism of claim 1 and isolating lipid from the microorganism, the culture medium, or both.
17. A method of producing lipid, comprising culturing a microorganism according to claim 1 under conditions in which the FAME to TOC ratio of the microorganism is maintained between 0.3 and 0.9, and isolating lipid from the microorganism, the culture medium, or both.
18. A method according to claim 17 , wherein the FAME to TOC ratio is maintained between about 0.4 and about 0.8.
19. A method of producing lipid, comprising culturing a mutant algal or heterokont microorganism that produces at least 30% more fatty acid methyl ester-derivatizable lipids (FAME lipids) than a substantially identical control algal or heterokont microorganism of the same species and at least 45% of the amount of biomass produced by the control microorganism when the mutant microorganism and control microorganism are cultured under identical conditions under which the control microorganism is producing biomass, wherein the mutant microorganism has attenuated expression of a gene encoding a polypeptide comprising an amino acid sequence having at least 80% identity to an amino acid sequence selected from the group consisting of: SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24.
20. The method of claim 19 , wherein the algal microorganism is a Eustigmatophyte.
21. A mutant algal or heterokont microorganism of claim 1 , having attenuated expression of a gene encoding a polypeptide comprising an amino acid sequence having at least 90% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24.
22. A mutant algal or heterokont microorganism of claim 19 , having attenuated expression of a gene encoding a polypeptide comprising an amino acid sequence having at least 90% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24.
23. A mutant algal or heterokont microorganism of claim 1 , wherein the culturing under identical conditions comprises culturing in nitrate or urea.
24. A mutant algal or heterokont microorganism of claim 19 , wherein the culturing under identical conditions comprises culturing in nitrate or urea.
25. A method of producing lipid, comprising culturing the microorganism of claim 22 , and isolating lipid from the microorganism, the culture medium, or both.
26. The method of claim 25 , wherein the algal microorganism is a Eustigmatophyte.