Process for producing peptide

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/62 (2006.01) C07K 14/58 (2006.01) C07K 14/585 (2006.01) C12N 9/38 (2006.01)

Patent

CA 2076320

A process for production of peptide (target peptide) comprising: A) culturing host cells transformed with plasmid able to express a gene coding for a fusion protein represented in the following formula: A-L-B wherein, B is a target peptide, A is a protective peptide which is a fragment of 90-210 amino acids from the N-terminal of E. coli-.beta.-galactosidase polypeptide, and L is a linker peptide positioned between the C-terminal of said protective peptide and the N- terminal of said target peptide and selected so that said fusion protein is treated by an enzyme or chemical substance, the above-mentioned target peptide is separated, and said linker peptide has basic amino acid residues selected so that the isoelectric point of the entire fusion protein A-L-B is adjusted to a range between 4.9 and 6.9; B) obtaining an insoluble fraction comprising inclusion bodies by homogenization of the cultured cells of said transform; C) solubilizing a fusion protein in said inclusion bodies by treatment of said insoluble fraction with solubilizing agent; and D) cleaving the peptide bond between the C-terminal of the linker amino acid residue and the N- terminal of the target peptide of said solubilized fusion protein to release said target peptide. The present invention is able to express a target peptide in a large amount and accumulate said target peptide in host cells in the form of inclusion bodies.

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