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US 2003/0143680 A1Published application
Descendants of bacteria devoid of N terminal formylation useful for the production of proteins and peptides
OwnerPhilippe Marliere, Rupert Mutzel +1inventors
InventorsPhilippe Marliere, Rupert Mutzel, Didier Mazel
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Abstract
The present invention relates to producing non-formylated proteins and/or peptides in bacterial cells lacking deformylase and/or transformylase.
Claims15
1:
- use of a bacterial strain lacking of deformylase and/or transformylase activities for the obtention of non formylated peptides or proteins.
2:
- use of a bacterial strain lacking of polypeptide deformylase and/or Met-tRNA transformylase activities for the obtention of non formylated peptides or proteins.
3:
- use of a bacterial strain according to claims 1 to 2 lacking of deformylase and/or transformylase activities characterized in that it is non reversible for the deformylase and/or transformylase activities
4:
- use of the bacterial strain according to claim 1 to 3 characterized in that it is cultivated at least at a temperature of 37° C.
5:
- use of a bacterial strain according to claim 1 and 4 characterized in that the bacteria is E. coli
6:
- A bacterial strain lacking of the deformylation and/or transformylation activity characterized in that
- said bacteria is not able to revert spontaneousely for these activities, and
- the growth rate of said bacteria is at least equivalent to the wild type growth rate.
7:
- use of the bacteria according to claim 6 for the production of non formylated peptides or proteins.
8:
- A process for producing peptides or proteins comprising the following steps:
- a) culture of the bacteria according to claim 6
- b) transforming the bacteria according to step a) with a DNA molecule or a plasmid or a vector comprising an insert containing a polynucleotide coding for an heterologous or homologous peptide or protein
- c) production of the peptide or the protein by the bacteria
- d) optionaly, separation of the peptide or protein of interest from the bacterial culture.
2 more elements
9:
- a purified polynucleotide comprising a gene coding for a mutator allele and contained in the bacteria strain according to claim 6
10:
- a purified polynucleotide according to claim 9 in which the mutator phenotype has been complemented or the mutator allele has been replaced by the wild type allele
11:
- process according to claim 8 in which the bacteria comprises a polynucleotide coding for a mutator allele.
12:
- Process according to claim 8 in which the bacteria mutator phenotype has been complemented or the mutator allele has been replaced by the wid type allele
13:
- E. coli bacterial strain γ2045 culture deposit at the CNCM (accesion number=I-2694 on Jul. 5, 2001 )
14:
- A process for the selection of mutants of a bacteria according to claim 6 characterized by the culture under continuous proliferation conditions of said mutants and separation of the mutant of interest from the static bacteria
15:
- Purified protein obtained after the expression of the heterologous insert of interest and deprived of formyl residue