Pufa polyketide synthase systems and uses thereof
Disclosed are the complete polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) systems from the bacterial microorganisms Shewanella japonica and Shewanella olleyana , and biologically active fragments and homologues thereof. More particularly, this invention relates to nucleic acids encoding such PUFA PKS systems, to proteins and domains thereof that comprise such PUFA PKS systems, to genetically modified organisms (plants and microorganisms) comprising such PUFA PKS systems, and to methods of making and using the PUFA PKS systems disclosed herein. This invention also relates to genetically modified plants and microorganisms and methods to efficiently produce lipids enriched in various polyunsaturated fatty acids (PUFAs) as well as other bioactive molecules by manipulation of a PUFA polyketide synthase (PKS) system.
1. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to amino acid positions 84-497 of SEQ ID NO:5 or that is an enzymatically active fragment of amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
2. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
3. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 95% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
4. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding an amino acid sequence that is at least 96% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
5. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 96% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
6. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding an amino acid sequence that is at least 97% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
7. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 97% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
8. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding an amino acid sequence that is at least 98% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
9. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 98% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
10. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding an amino acid sequence that is at least 99% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
11. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding an amino acid sequence that is at least 99% identical to amino acid positions 84-497 of SEQ ID NO:5, wherein said amino acid sequence has enoyl reductase (ER) activity.
12. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence encoding amino acid positions 84-497 of SEQ ID NO:5.
13. The isolated nucleic acid molecule of claim 1 , consisting of a nucleic acid sequence encoding amino acid positions 84-497 of SEQ ID NO:5.
14. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises nucleotides 27446-28687 of SEQ ID NO:1.
15. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule consists of nucleotides 27446-28687 of SEQ ID NO:1.
16. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises a nucleic acid sequence encoding SEQ ID NO:5.
17. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO:1.
18. A recombinant nucleic acid molecule comprising the nucleic acid molecule of claim 1 and a transcription control sequence.
19. A recombinant plant cell that expresses the nucleic acid molecule of claim 1 .
20. A recombinant microbial cell that expresses a recombinant vector comprising the nucleic acid molecule of claim 1 and a transcription control sequence.
21. The recombinant microbial cell of claim 20 , wherein the microbial cell is a bacterium.
22. The recombinant microbial cell of claim 20 , wherein the microbial cell is a Thraustochytriales microorganism.
23. The recombinant microbial cell of claim 22 , wherein the Thraustochytriales microorganism is a Schizochytrium or a Thraustochytrium.
24. A method to produce at least one polyunsaturated fatty acid (PUFA), comprising culturing under conditions effective to produce the PUFA, a plant cell or a microbial cell that expresses a PKS system for production of PUFAs, wherein the plant cell or microbial cell expresses a recombinant vector comprising the nucleic acid molecule of claim 1 .