# US 5,702,922 A · Method for the extraction of glicentin or glicentin analogous substances > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,702,922 A - Document ID: US5702922A - Status: Granted - Kind: A - Application number: 08/562,148 - Filed: 1995-11-22 - Granted: 1997-12-30 - Earliest claimed: 1994-11-29 - Assignees: Nisshin Flour Milling Co., Ltd. - Inventors: Shinjirou Imai - Web page: https://patentgrep.com/patent/US5702922A ## CPC - C07K 14/605: Glucagons - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract In the method for the extraction of glicentin or a glicentin analogous substance from the cells of a transformed microorganism having incorporated therein an expressible gene encoding glicentin or a glicentin analogous substance, the extraction is carried out using an aqueous solution of an inorganic acid and/or an organic acid. The glicentin is human glicentin or a human glicentin analogous substance. ## Claims ### Claim 1 A method for the extraction of glicentin or a glicentin analogous substance from the cells of a transformed microorganism having incorporated therein an expressible gene encoding glicentin or the glicentin analogous substance, which comprises: a) extracting said glicentin or glicentin analogous substance from said cells with an aqueous acidic solution; and b) isolating said glicentin or said glicentin analogous substance. ### Claim 2 (depends on 1) The method of claim 1, wherein said glicentin is human glicentin and said glicentin analogous substance is a human glicentin analogous substance. ### Claim 3 (depends on 1) The method of claim 1, wherein the glicentin analogous substance is a peptide comprising 29 or more amino acid residues of an entire amino acid sequence of glicentin. ### Claim 4 (depends on 1) The method of claim 1, wherein the aqueous acid solution is an aqueous solution comprising an inorganic acid or an organic acid or both. ### Claim 5 (depends on 4) The method of claim 4, wherein the inorganic acid is selected from the group consisting of hydrochloric acid and phosphoric acid. ### Claim 6 (depends on 4) The method of claim 4, wherein the aqueous acid solution has a pH of not more than 3 and the inorganic acid has a concentration of from about 10 to 1,000 mM. ### Claim 7 (depends on 4) The method of claim 4, wherein the organic acid is acetic acid. ### Claim 8 (depends on 4) The method of claim 4, wherein the aqueous acid solution has a pH of not more than 3 and the organic acid has a concentration of about 5 to 20% (w/w). ### Claim 9 (depends on 1) The method of claim 1, wherein said transformed microorganism is a gram-negative bacterium. ### Claim 10 (depends on 9) The method of claim 9, wherein said transformed microorganism is E. coli. ### Claim 11 (depends on 1) The method of claim 1, wherein said glicentin analogous substance comprises a sequence composed of the 1st to 61st amino acid residues of glicentin, a sequence composed of the 1st to 30th amino acid residues of glicentin, a sequence composed of the 33rd to 69th amino acid residues of glicentin or a sequence composed of the 33rd to 61th amino acid residues of glicentin. ### Claim 12 (depends on 1) The method of claim 1, wherein said extraction is effected at about 0.degree. to 37.degree. C. ### Claim 13 (depends on 12) The method of claim 12, wherein said extraction is effected at about 0.degree. to 10.degree. C. ### Claim 14 A method for the extraction of glicentin or a glicentin analogous substance from the cells of a transformed microorganism having incorporated therein an expressible gene encoding the glicentin or the glicentin analogous substance, which comprises: a) pretreating the cells of the transformed microorganism with an aqueous acid solution; b) separating cell debris and extracting said glicentin or said glicentin analogous substance with the aqueous acid solution; and c) isolating said glicentin or said glicentin analogous substance. ### Claim 15 (depends on 14) The method of claim 14, wherein said glicentin is human glicentin and said glicentin analogous substance is a human glicentin analogous substance. ### Claim 16 (depends on 14) The method of claim 14, wherein the glicentin analogous substance is a peptide comprising 29 or more amino acid residues of an entire amino acid sequence of glicentin. ### Claim 17 (depends on 14) The method of claim 14, wherein the aqueous acid solution is an aqueous solution comprising an inorganic acid or an organic acid or both. ### Claim 18 (depends on 17) The method of claim 17, wherein the inorganic acid is selected from the group consisting of hydrochloric acid and phosphoric acid. ### Claim 19 (depends on 17) The method of claim 17, wherein the aqueous acid solution has a pH of not more than 3 and the inorganic acid has a concentration of from about 10 to 1,000 mm. ### Claim 20 (depends on 17) The method of claim 17, wherein the organic acid is acetic acid. ### Claim 21 (depends on 17) The method of claim 17, wherein the aqueous acid solution has a pH of not more than 3 and the organic acid has a concentration of about 5 to 20% (w/w). ### Claim 22 (depends on 14) The method of claim 14, wherein said transformed microorganism is a gram-negative bacterium. ### Claim 23 (depends on 22) The method of claim 22, wherein said transformed microorganism is E. coli. ### Claim 24 (depends on 14) The method of claim 14, wherein said glicentin analogous substance comprises a sequence-composed of the 1st to 61st amino acid residues of glicentin, a sequence composed of the 1st to 30th amino acid residues of glicentin, a sequence composed of the 33rd to 69th amino acid residues of glicentin or a sequence composed of the 33rd to 61th amino acid residues of glicentin. ### Claim 25 (depends on 14) The method of claim 14, wherein said extraction is effected at about 0.degree. to 37.degree. C. ### Claim 26 (depends on 25) The method of claim 25, wherein said extraction is effected at about 0.degree. to 10.degree. C.