Abstract

Escherichia coli of which a redox potential is not adjusted, and a CRM197 protein having high periplasmic secretion efficiency may be produced even without shifting the pH of a culture medium in order to increase secretion into the periplasm, and thus the present invention is very useful in CRM197 protein production.

IPC Classes  ?

• C12P 21/02 - Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
• A61K 39/05 - CorynebacteriumPropionibacterium
• C07K 7/08 - Linear peptides containing only normal peptide links having 12 to 20 amino acids
• C07K 14/34 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from bacteria from Corynebacterium (G)

Abstract

The present invention relates to a signal sequence for expressing a CRM197 protein in Escherichia coli and secreting same into the periplasm, and a use thereof, and more specifically, to: a signal sequence for expressing a CRM197 protein; a nucleic acid for coding the signal sequence; a nucleic acid construct or expression vector comprising the nucleic acid and a CRM197 protein gene; a recombinant microorganism having the nucleic acid construct or expression vector introduced therein; and a CRM197 protein production method comprising a step for culturing the recombinant microorganism. According to the present invention, a CRM197 protein having the same physicochemical/immunologic properties as the protein isolated from the parent bacteria may be expressed even in regular Escherichia coli of which a redox potential is not adjusted, and a CRM197 protein having high periplasmic secretion efficiency may be produced even without shifting the pH of a culture medium in order to increase secretion into the periplasm, and thus the present invention is very useful in CRM197 protein production.

IPC Classes  ?

• C07K 7/08 - Linear peptides containing only normal peptide links having 12 to 20 amino acids
• C07K 14/00 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof
• C07K 14/34 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from bacteria from Corynebacterium (G)

Abstract

The present invention relates to a signal sequence for expressing a CRM197 protein in Escherichia coli and secreting same into the periplasm, and a use thereof, and more specifically, to: a signal sequence for expressing a CRM197 protein; a nucleic acid for coding the signal sequence; a nucleic acid construct or expression vector comprising the nucleic acid and a CRM197 protein gene; a recombinant microorganism having the nucleic acid construct or expression vector introduced therein; and a CRM197 protein production method comprising a step for culturing the recombinant microorganism. According to the present invention, a CRM197 protein having the same physicochemical/immunologic properties as the protein isolated from the parent bacteria may be expressed even in regular Escherichia coli of which a redox potential is not adjusted, and a CRM197 protein having high periplasmic secretion efficiency may be produced even without shifting the pH of a culture medium in order to increase secretion into the periplasm, and thus the present invention is very useful in CRM197 protein production.

IPC Classes  ?

• C07K 7/08 - Linear peptides containing only normal peptide links having 12 to 20 amino acids
• C07K 14/34 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from bacteria from Corynebacterium (G)

Abstract

Corynebacteriumdiphtheriaediphtheriae EB01 (KCTC13549BP) which expresses a high concentration of CRM197, according to the present invention, is a double lysogenic strain with which the expression of CRM197 is 1.5 times to 3 times greater than when a single lysogenic strain is used, and thus the time and cost required for production can be reduced. Since the Corynebacterium strain can express high concentrations of CRM197 without being affected by the concentration of Fe 2+, a step for removing Fe 2+is unnecessary during the preparation of a culture medium, and bacterial growth can be improved under conditions of high Fe 2+ concentration without influencing the expression of CRM197, and as a result, the productivity of CRM197 can be improved. Ultimately, this enables production of a high yield of CRM197 through simpler processes, such that manufacturing costs are reduced, and thus it is possible to develop an economical conjugate vaccine production process.

IPC Classes  ?

• C12N 15/77 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for CorynebacteriumVectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Brevibacterium
• C12N 15/01 - Preparation of mutants without inserting foreign genetic material thereinScreening processes therefor
• C12N 1/20 - BacteriaCulture media therefor
• C07K 14/34 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof from bacteria from Corynebacterium (G)
• C12R 1/16 - Corynebacterium diphtheriae