Fusion moieties and microbial hosts for protein production
The present invention relates to fusion proteins comprising (i) a fusion moiety based on SEQ ID NO:1 and (ii) a protein. Also provided are nucleic acids encoding such fusion proteins and compositions comprising such fusion proteins. The invention also provides a method for increasing the expression level of a protein in a host cell or increasing the level of secretion of a protein from a host cell, said methods employing a fusion moiety in accordance with the invention. The invention further provides a method of producing a protein, said method comprising culturing an Aliivibrio wodanis host cell comprising a heterologous nucleic acid molecule encoding a protein under conditions suitable for the expression of the encoded protein. Certain deposited strains of Aliivibrio wodanis are also provided.
1. A nucleic acid molecule encoding a fusion protein comprising
(i) a fusion moiety comprising the amino acid sequence of SEQ ID NO:1 or an amino acid sequence that is at least 80% identical to SEQ ID NO:1; and
(ii) a protein.
2. A nucleic acid molecule encoding a fusion protein, said nucleic acid molecule comprising
(i) a polynucleotide comprising the polynucleotide sequence of SEQ ID NO:2 or SEQ ID NO:5, or a polynucleotide sequence that is at least 80% identical to SEQ ID NO:2 or SEQ ID NO:5; and
(ii) a polynucleotide encoding a protein.
3. A method of producing a protein, said method comprising culturing a host cell comprising a heterologous nucleic acid molecule encoding said protein under conditions suitable for the expression of the encoded protein, wherein said protein is a fusion protein comprising
(i) a fusion moiety comprising the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:4 or an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or SEQ ID NO:4, and
(ii) a protein.
4. The method of claim 3 , said method further comprising, subsequent to culturing the host cell, a step of isolating the protein from the host cell or from the growth medium or supernatant.
5. The method of claim 3 , wherein said host cell is an Aliivibrio wodanis host cell.
6. A method of producing a protein, said method comprising culturing an Aliivibrio wodanis host cell comprising a heterologous nucleic acid molecule encoding said protein under conditions suitable for the expression of the encoded protein, wherein said protein is a fusion protein comprising
(i) a fusion moiety comprising the amino acid sequence of SEQ ID NO:1 or an amino acid sequence that is at least 80% identical to SEQ ID NO:1; and
(ii) a protein.
7. The method of claim 6 , wherein said Aliivibrio wodanis is an Aliivibrio wodanis strain selected from the group consisting of:
(i) the Aliivibrio wodanis strain deposited under accession number ECACC 18050101;
(ii) the Aliivibrio wodanis strain deposited under accession number ECACC 18050102; and
(iii) an Aliivibrio wodanis strain having all the identifying characteristics of one or both of strains (i) and (ii).
8. The method of claim 6 , wherein the culturing is done at a temperature of 4° C. to 18° C.
9. The method of claim 6 , said method (i) further comprising, prior to culturing the Aliivibrio wodanis , a step of introducing said heterologous nucleic acid molecule encoding said protein into the Aliivibrio wodanis host cell or (ii) further comprising, subsequent to culturing the Aliivibrio wodanis , a step of isolating the protein from the Aliivibrio wodanis or from the growth medium or supernatant.
10. An Aliivibrio wodanis strain selected from the group consisting of:
(i) the Aliivibrio wodanis strain deposited under accession number ECACC 18050101;
(ii) the Aliivibrio wodanis strain deposited under accession number ECACC 18050102; and
(iii) an Aliivibrio wodanis strain having all the identifying characteristics of one or both of strains (i) and (ii),
wherein the Aliivibrio wodanis strain comprises a heterologous nucleic acid molecule encoding a protein.
11. The method of claim 6 , said method (i) further comprising, prior to culturing the Aliivibrio wodanis , a step of introducing said heterologous nucleic acid molecule encoding said protein into the Aliivibrio wodanis host cell and (ii) further comprising, subsequent to culturing the Aliivibrio wodanis , a step of isolating the protein from the Aliivibrio wodanis or from the growth medium or supernatant.
12. The method of claim 4 further comprising the step of formulating the protein into a composition including at least one additional component.
13. The method of claim 9 further comprising the step of formulating the protein into a composition including at least one additional component.
14. The method of claim 11 further comprising the step of formulating the protein into a composition including at least one additional component.