# US 10,870,871 B2 · Polypeptide and method for producing ornithine-based product using the same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,870,871 B2 - Document ID: US10870871B2 - Status: Granted - Kind: B2 - Application number: 16/622,087 - Filed: 2018-06-14 - Published: 2020-06-11 - Granted: 2020-12-22 - Earliest claimed: 2017-06-14 - Assignees: Cj Cheiljedang Corporation - Applicants: Cj Cheiljedang Corporation - Inventors: Seon Hye Kim; Su Jin Park; Kyoung Min Lee; Kyungsu Na; Hong Xian Li; Hyun-jung Bae; Jihyun Shim; Young Lyeol Yang; Hye Won Um; Hyo Hyoung Lee; Min Gyeong Kang; Hye Won Kim; Byeong Cheol Song; Haena Oh; Han Hyoung Lee - Web page: https://patentgrep.com/patent/US10870871B2 ## CPC - C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} - C12P 13/10: Citrulline; Arginine; Ornithine - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - C12N 15/77: for Corynebacterium; for Brevibacterium - C12P 13/001: {Amines; Imines} - 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) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present disclosure relates to a novel polypeptide having an ability to export an ornithine-based product, and a method for producing an ornithine-based product using the same. ## Claims ### Claim 1 A polypeptide having putrescine or arginine export activity and comprising an amino acid sequence that has at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, and wherein the alanine at the position corresponding to residue 152 of SEQ ID NO: 1 or SEQ ID NO: 2 is substituted with a different amino acid in the amino acid sequence of the polypeptide. ### Claim 2 (depends on 1) The polypeptide of claim 1, wherein the alanine at the position corresponding to residue 152 of SEQ ID NO: 1 or SEQ ID NO: 2 is substituted with serine, asparagine, histidine, proline, lysine, glutamic acid, cysteine, glutamine, or methionine. ### Claim 3 (depends on 1) The polypeptide of claim 1, wherein the polypeptide consists of the amino acid sequence of any one of SEQ ID NO: 3 to SEQ ID NO: 20. ### Claim 4 (depends on 1) A polynucleotide comprising a nucleotide sequence encoding the polypeptide of claim 1. ### Claim 5 (depends on 4) A vector comprising the polynucleotide of claim 4. ### Claim 6 (depends on 1) A microorganism of the genus Corynebacterium, wherein the microorganism produces putrescine and comprises the polypeptide of claim 1. ### Claim 7 (depends on 6) The microorganism of claim 6, wherein the microorganism is Corynebacterium glutamicum. ### Claim 8 (depends on 6) The microorganism of claim 6, wherein the microorganism further comprises an ornithine decarboxylase (ODC). ### Claim 9 (depends on 6) The microorganism of claim 6, wherein a gene encoding at least one polypeptide selected from the group consisting of ornithine carbamoyltransferase and a protein involved in glutamate export is inactivated. ### Claim 10 (depends on 6) The microorganism of claim 6, wherein expression of a polynucleotide encoding at least one polypeptide selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC), acetylglutamate synthase or ornithine acetyltransferase (argJ), acetylglutamate kinase (ArgB), and acetylornithine aminotransferase (ArgD) is enhanced compared to a corresponding wild-type microorganism of the genus Corynebacteriumthc. ### Claim 11 (depends on 6) The microorganism of claim 6, wherein a gene encoding putrescine acetyltransferase is inactivated. ### Claim 12 (depends on 1) A microorganism of the genus Corynebacterium, wherein the microorganism produces arginine and comprises the polypeptide of claim 1, and wherein the arginine-producing ability of the microorganism is enhanced compared to a corresponding microorganism of the genus Corynebacterium that does not comprise the polypeptide of claim 1. ### Claim 13 (depends on 12) The microorganism of claim 12, wherein expression of a polynucleotide encoding at least one polypeptide selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC), acetylglutamate synthase or ornithine acetyltransferase (argJ), acetylglutamate kinase (ArgB), and acetylornithine aminotransferase (ArgD) is enhanced compared to a corresponding wild-type microorganism of the genus Corynebacterium. ### Claim 14 (depends on 12) The microorganism of claim 12, wherein expression of a polynucleotide encoding at least one polypeptide selected from the group consisting of ornithine carbamoyltransfrase (ArgF), argininosuccinate synthase (argG), argininosuccinate lyase (argH), aspartate ammonia lyase and aspartate aminotransferase is increased compared to a corresponding wild-type microorganism of the genus Corynebacterium. ### Claim 15 (depends on 12) The microorganism of claim 12, wherein the microorganism is Corynebacterium glutamicum. ### Claim 16 (depends on 6) A method for producing putrescine, comprising: (i) culturing the microorganism of claim 6 in a medium; and (ii) recovering putrescine from the microorganism or the medium. ### Claim 17 (depends on 12) A method for producing arginine, comprising: (i) culturing the microorganism of claim 12 in a medium; and (ii) recovering arginine from the microorganism or the medium. ### Claim 18 (depends on 1) A microorganism, wherein the microorganism produces putrescine or arginine and comprises the polypeptide of claim 1. ### Claim 19 (depends on 18) The microorganism of claim 18, wherein expression of a polynucleotide encoding at least one polypeptide selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC), acetylglutamate synthase or ornithine acetyltransferase (argJ), acetylglutamate kinase (ArgB), and acetylornithine aminotransferase (ArgD) is enhanced compared to a corresponding wild-type microorganism of the genus Corynebacterium. ### Claim 20 (depends on 18) The microorganism of claim 18, wherein the microorganism is Corynebacterium glutamicum. ### Claim 21 (depends on 18) A method for producing putrescine or arginine, comprising: (i) culturing the microorganism of claim 18 in a medium; and (ii) recovering putrescine or arginine from the microorganism or the medium.