Protein production in microorganisms of the phylum labyrinthulomycota
View Patent ↗The present invention relates to recombinant cells and microorganisms of the phylum Labyrinthulomycota and their use in heterologous protein production. Novel promoter, terminator, and signal sequences for efficient production and, optionally, secretion of polypeptides from recombinant host cells and microorganisms are also encompassed by the present invention.
1. An isolated recombinant nucleic acid molecule comprising:
(a) a polynucleotide sequence having at least 90% sequence identity to the polynucleotide sequence of SEQ ID NO:38, wherein the polynucleotide sequence encodes a polypeptide that functions as a signal peptide;
(b) a polynucleotide sequence encoding a polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:37 or a fragment thereof, wherein the amino acid sequence or fragment thereof functions as a signal peptide; or
(c) a polynucleotide sequence that is fully complementary to the polynucleotide sequence of (a) or (b).
2. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence has at least 95% sequence identity to the polynucleotide sequence of SEQ ID NO:38.
3. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence is SEQ ID NO:38.
4. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:37.
5. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:37.
6. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence encodes a polypeptide comprising the first 18 amino acid residues of SEQ ID NO:37.
7. The recombinant nucleic acid molecule of claim 1 , wherein the polynucleotide sequence encodes a polypeptide comprising the first 19 amino acid residues of SEQ ID NO:37.
8. The recombinant nucleic acid molecule of claim 1 , wherein the recombinant nucleic acid molecule is a vector.
9. The recombinant nucleic acid molecule of claim 1 wherein the polynucleotide sequence is operably linked to a polynucleotide sequence encoding a protein.
10. A host cell comprising the recombinant nucleic acid molecule of claim 1 .
11. The host cell of claim 10 , wherein the host cell is a member of the order Thraustochytriales.
12. The host cell of claim 11 , wherein the host cell is a Schizochytrium or a Thraustochytrium.
13. A method for production of a protein encoded by the recombinant nucleic acid molecule of claim 9 , comprising:
a) culturing a recombinant microorganism of the order Thraustochytriales in a medium, wherein the recombinant microorganism comprises the isolated nucleic acid molecule of claim 9 ; and
b) isolating the protein made in step (a).
14. The method of claim 13 , wherein the protein is recovered from an isolated Thraustoehytriales biomass.
15. The method of claim 13 , wherein the protein accumulates in the microorganism.
16. The method of claim 13 , wherein the protein accumulates in a membrane of the microorganism.
17. The method of claim 13 , wherein the protein is recovered from the culture medium.
18. The method of claim 13 , wherein the protein is secreted.