Descendants of bacteria devoid of N terminal formylation useful for the production of proteins and peptides
View Patent ↗The present invention relates to producing non-formylated proteins and/or peptides in bacterial cells lacking deformylase and/or transformylase.
1. A bacterial cell which is Escherichia coli that has been deleted for the fmt (Met-tRNA transformylase) gene or both the def (deformylase) and fmt (Met-tRNA transformylase) genes from the def-fmt operon,
wherein said bacterial cell expresses non-formylated proteins, polypeptides or peptides.
2. The bacterial cell of claim 1 , in which SEQ ID NO: 4 (an E. coli fmt polynucleotide sequence) has been deleted.
3. The bacterial cell of claim 1 , in which both SEQ ID NO: 2 (an E. coli def polynucleotide sequence) and SEQ ID NO: 4 (an E. coli fmt polynucleotide sequence) have been deleted.
4. The bacterial cell of claim 1 , which lacks deformylase activity.
5. The bacterial cell of claim 1 , which lacks deformylase and Met-tRNA transformylase activity.
6. The bacterial cell of claim 1 , wherein the growth rate of said cell at 37° C. is at least equivalent to the growth rate of the corresponding bacterial cell not having said fmt or def-fmt deletion(s).
7. The bacterial cell of claim 1 wherein the growth rate of said cell at 42° C. is at least equivalent to the growth rate of the corresponding bacterial cell not having said fmt or def-fmt deletion(s).
8. The bacterial cell of claim 1 which is Escherichia coli strain γ2045 (CNCM I-2694).
9. The bacterial cell of claim 1 , further comprising an exogenous polynucleotide sequence encoding a protein, polypeptide or peptide.
10. The bacterial cell of claim 1 , further comprising a heterologous polynucleotide sequence encoding a protein, polypeptide or peptide.
11. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 1 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
12. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 2 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
13. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 3 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
14. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 4 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
15. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 5 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
16. The method of claim 11 , wherein said bacterial cell is cultivated at 37° C.
17. The method of claim 11 , wherein said bacterial cell is cultivated at 42° C.
18. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 8 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said non-formylated protein, polypeptide and/or peptide.
19. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 9 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said exogenous non-formylated protein, polypeptide and/or peptide.
20. A method of producing a non-formylated protein, polypeptide and/or peptide comprising:
cultivating the bacterial cell of claim 10 for a time and under conditions suitable for expression of a non-formylated protein, polypeptide and/or peptide, and, optionally,
recovering or isolating said heterologous non-formylated protein, polypeptide and/or peptide.