Process for production of physiologically active peptide...

C - Chemistry – Metallurgy – 12 – P

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530/13, 195/1.29

C12P 21/02 (2006.01) C07K 14/58 (2006.01) C07K 14/585 (2006.01) C07K 19/00 (2006.01) C12N 9/38 (2006.01) C12N 15/62 (2006.01) C12N 15/71 (2006.01)

Patent

CA 1340830

A process for the production of a physiologically active peptide (a target peptide) containing cysteine residue, comprising the steps of: (1) culturing Escherichia coli transformed with a plasmid capable of expressing a fusion protein under control of a promoter of E. coli origin or a promoter of a phage origin, wherein the fusion protein is represented by the following formula: A - L - B wherein B represents a target peptide containing cysteine residue; A represents a partner polypeptide consisting of 90 to 220 amino acid residue but not containing cysteine residue; and L represents a linker amino acid residue positioned between C-terminal of the partner polypeptide and N-terminal of the target peptide wherein the same amino acid as the linker amino acid is not present in the target peptide, and the linker amino acid is selected so that the peptide bond between the C-terminal of the linker amino acid and the N-terminal of the target peptide is claimed by a protease or the linker amino acid is selectively degraded by a chemical substance; (2) disrupting the cultured cells and obtaining an insoluble fraction containing the fusion protein; (3) solubilizing the fusion protein with a solubilizing agent, and treating the solubilized fusion protein with the protease or the chemical substance to liberate the target peptide, and isolating the target peptide.

560605

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