Enzymes for producing non-straight-chain fatty acids
US-2015376659-A1 · Dec 31, 2015 · US
US9909155B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9909155-B2 |
| Application number | US-201615369557-A |
| Country | US |
| Kind code | B2 |
| Filing date | Dec 5, 2016 |
| Priority date | Apr 18, 2012 |
| Publication date | Mar 6, 2018 |
| Grant date | Mar 6, 2018 |
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Recombinant DNA techniques are used to produce oleaginous recombinant cells that produce triglyceride oils having desired fatty acid profiles and regiospecific or stereospecific profiles. Genes manipulated include those encoding stearoyl-ACP desturase, delta 12 fatty acid desaturase, acyl-ACP thioesterase, ketoacyl-ACP synthase, and lysophosphatidic acid acyltransferase. The oil produced can have enhanced oxidative or thermal stability, or can be useful as a frying oil, shortening, roll-in shortening, tempering fat, cocoa butter replacement, as a lubricant, or as a feedstock for various chemical processes. The fatty acid profile can be enriched in midchain profiles or the oil can be enriched in triglycerides of the saturated-unsaturated-saturated type.
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What is claimed is: 1. A recombinant microalgal cell, the cell comprising exogenous nucleic acids encoding a C16:0 or C18:0 preferring acyl-ACP thioesterase A (FATA), wherein the oil produced by the recombinant microalgal cell is enriched in stearic-oleic-stearic (SOS), palmitic-oleic-palmitic (POP) and/or palmitic-oleic-stearic (POS) triglycerides, as compared to non-recombinant microalgal cells. 2. The recombinant microalgal cell of claim 1 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Brassica FATA or a Garcinia FATA. 3. The recombinant microalgal cell of claim 2 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Garcinia FATA. 4. The recombinant microalgal cell of claim 3 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Garcinia mangostana FATA. 5. The method of claim 2 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Brassica FATA. 6. The recombinant microalgal cell of claim 1 , wherein the recombinant microalgal cell further comprises nucleic acids that encode sucrose invertase. 7. The recombinant microalgal cell of claim 1 , wherein the oil produced by the recombinant microalgal cell comprises at least 50% SOS, or optionally at least 70% SOS. 8. The recombinant microalgal cell of claim 1 , wherein the microalgal cell is of the genus Prototheca or Chlorella. 9. The recombinant microalgal cell of claim 8 , wherein the microalgal cell is of the species Prototheca moriformis. 10. The recombinant microalgal cell of claim 1 , wherein the exogenous nucleic acids are codon optimized for expression in a cell of the genus Prototheca. 11. A method of producing a microalgal oil enriched in stearic-oleic-stearic (SOS), palmitic-oleic-palmitic (POP) and/or palmitic-oleic-stearic (POS) triglycerides, as compared to non-recombinant microalgal cells, the method comprising the steps of: a) providing a recombinant microalgal cell, the cell comprising exogenous nucleic acids encoding a C16:0 or C18:0 preferring acyl-ACP thioesterase A (FATA); b) cultivating the recombinant microalgal cell; and c) isolating the microalgal oil from the microalgal cell. 12. The method of claim 11 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Brassica FATA or a Garcinia FATA. 13. The method of claim 12 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Garcinia FATA. 14. The method of claim 3 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Garcinia mangostana FATA. 15. The method of claim 12 , wherein the C16:0 or C18:0 preferring acyl-ACP thioesterase A is a Brassica FATA. 16. The recombinant microalgal cell of claim 11 , wherein the recombinant microalgal cell further comprises nucleic acids that encode sucrose invertase. 17. The method of claim 11 , wherein the oil produced by the recombinant microalgal cell comprises at least 50% SOS, or optionally at least 70% SOS. 18. The method of claim 11 , wherein the microalgal cell is of the genus Prototheca or Chlorella. 19. The method of claim 18 , wherein the microalgal cell is of the species Prototheca moriformis. 20. The method of claim 11 , wherein the exogenous nucleic acids are codon optimized for expression in a cell of the genus Prototheca.
Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title
Beta-ketoacyl-acyl-carrier-protein synthase III (2.3.1.180) · CPC title
Other edible oils or fats, e.g. shortenings or cooking oils (preservation of other edible oils A23B20/30) · CPC title
Fatty acids · CPC title
Glycerol-3-phosphate O-acyltransferase (2.3.1.15) · CPC title
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