Polyunsaturated fatty acid synthase nucleic acid molecules and polypeptides, compositions, and methods of making and uses thereof
The present invention is directed to isolated nucleic acid molecules and polypeptides of thraustochytrid polyunsaturated fatty acid (PUFA) synthases involved in the production of PUFAs, including PUFAs enriched in docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a combination thereof. The present invention is directed to vectors and host cells comprising the nucleic acid molecules, polypeptides encoded by the nucleic acid molecules, compositions comprising the nucleic acid molecules or polypeptides, and methods of making and uses thereof.
1. An recombinant nucleic acid molecule comprising a polynucleotide sequence encoding a polypeptide and a transcription control sequence, wherein the polypeptide comprises an amino acid sequence at least 90% identical to SEQ ID NO:8, wherein the polypeptide comprises KS activity, said polynucleotide sequence is heterologous to said transcription control sequence.
2. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence is at least 95% identical to SEQ ID NO:8.
3. A host cell that expresses any one of the nucleic acid molecule of claim 1 , wherein said nucleic acid molecule is heterologous to the host cell, and wherein said host cell is not a human cell.
4. The host cell of claim 3 , wherein the host cell is selected from the group consisting of a plant cell, a microbial cell, and an isolated animal cell.
5. The host cell of claim 4 , wherein the microbial cell is a thraustochytrid, excluding Thraustochytrium sp. ATCC PTA-9695.
6. The host cell of claim 5 , wherein the thraustochytrid is a Schizochytrium or a Thraustochytrium.
7. The host cell of claim 4 , wherein the plant cell is selected from the group consisting of: canola, soybean, rapeseed, linseed/flax, maize, safflower, sunflower, tobacco, Arabidopsis thaliana, Brazil nut, castor bean, coconut, coriander, cotton, groundnut, jojoba, mustard, oil palm, olive, rice, squash, barley, wheat, and duckweed.
8. A method to produce at least one PUFA, comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce PUFA, wherein the PUFA synthase gene comprises any one of the nucleic acid molecule of claim 1 , and wherein at least one PUFA is produced, and wherein the nucleic acid molecule is heterologous to the host cell.
9. A method to produce lipids enriched for DHA, EPA, or a combination thereof, comprising: expressing a PUFA synthase gene in a host cell under conditions effective to produce lipids, wherein the PUFA synthase gene comprises the nucleic acid molecule of claim 1 , and wherein lipids enriched with DHA, EPA, or a combination thereof are produced.
10. A method of increasing production of DHA, EPA, or a combination thereof in an organism having PUFA synthase activity, comprising: expressing the recombinant nucleic acid molecule of claim 1 in the organism under conditions effective to produce DHA, EPA, or a combination thereof, wherein said nucleic acid molecule is heterologous to the host organism, and wherein the PUFA synthase activity replaces an inactive or deleted activity, introduces a new activity, or enhances an existing activity in the organism, and wherein production of DHA, EPA, or a combination thereof in the organism is increased.