# US 2020/0399671 A1 · Fusion protein of flavin adenine dinucleotide-glucose dehydrogenase and cytochrome molecule > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399671 A1 - Document ID: US20200399671A1 - Status: Published application - Kind: A1 - Application number: 16/979,161 - Filed: 2019-03-08 - Published: 2020-12-24 - Earliest claimed: 2018-03-08 - Assignees: Ultizyme International Ltd.; I-Sens, Inc. - Applicants: Ultizyme International Ltd.; I-Sens, Inc. - Inventors: Koji SODE; Youngjea KANG; Kazushige MORI; Katsuhiro KOJIMA; Junko SHIMAZAKI - Web page: https://patentgrep.com/patent/US20200399671A1 ## CPC - C12N 15/70: Vectors or expression systems specially adapted for E. coli - C12Q 1/006: {for glucose} - C07K 14/245: Escherichia (G) - C07K 14/80: Cytochromes - C12N 9/0006: {acting on CH-OH groups as donors (1.1)} - C12N 9/0008: {acting on the aldehyde or oxo group of donors (1.2)} - C12N 11/02: Enzymes or microbial cells immobilised on or in an organic carrier - C12N 15/62: DNA sequences coding for fusion proteins - C12Q 1/32: involving dehydrogenase - C07K 2319/00: Fusion polypeptide - C12Y 101/05: with a quinone or similar compound as acceptor (1.1.5) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - 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) - C12Y: ENZYMES ## Abstract The purpose of the present invention is to provide a fungus-derived FADGDH that has a direct electron transfer ability. Provided is a fusion protein comprising a fungus-derived FADGDH or a mutant thereof and a cytochrome molecule connected to the N-terminus of the FADGDH or variant thereof. ## Claims ### Claim 1 A fusion protein comprising: a fungus-derived flavin adenine dinucleotide glucose dehydrogenase (FADGDH) or a variant thereof; and a cytochrome molecule linked to the N-terminus of the FADGDH or variant thereof. ### Claim 2 (depends on 1) The fusion protein of claim 1, wherein the FADGDH is an FADGDH derived from Aspergillus sp., Botryotinia sp., Mucor sp., Parasitella sp., or Rhizopus sp. ### Claim 3 (depends on 1) The fusion protein of claim 1, wherein the cytochrome molecule is derived from E. coli or oxidoreductase. ### Claim 4 (depends on 1) The fusion protein of claim 1, wherein the cytochrome molecule is cytochrome b. ### Claim 5 (depends on 1) The fusion protein of claim 1, which is one of the following (a) to (c): (a) a protein which consists of an amino acid sequence set forth in at least one of SEQ ID NOs: 2, 3 and 13 to 21; and (b) a protein which consists of an amino acid sequence resulting from deletion, substitution, insertion or addition of one amino acid or a plurality of amino acids in the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function; and (c) a protein which consists of an amino acid sequence having a sequence identity of 80% or higher to the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function. ### Claim 6 (depends on 1) A gene encoding the fusion protein set forth in claim 1. ### Claim 7 (depends on 6) A vector comprising the gene set forth in claim 6. ### Claim 8 (depends on 6) A transformant comprising the gene set forth in claim 6. ### Claim 9 (depends on 6) A transformant in which the gene set forth in claim 6 is integrated into the main chromosome of the transformant. ### Claim 10 (depends on 1) An enzyme electrode comprising the fusion protein set forth in claim 1 attached thereto. ### Claim 11 (depends on 10) A method for measuring glucose concentration in a sample, the method comprising: bringing the sample into contact with the enzyme electrode set forth in claim 10; and measuring electrons generated by oxidation of glucose. ### Claim 12 (depends on 10) A glucose sensor which uses the enzyme electrode set forth in claim 10 as a working electrode. ### Claim 13 (depends on 2) The fusion protein of claim 2, wherein the cytochrome molecule is derived from E. coli or oxidoreductase. ### Claim 14 (depends on 2) The fusion protein of claim 2, wherein the cytochrome molecule is cytochrome b. ### Claim 15 (depends on 3) The fusion protein of claim 3, wherein the cytochrome molecule is cytochrome b. ### Claim 16 (depends on 2) The fusion protein of claim 2, which is one of the following (a) to (c): (a) a protein which consists of an amino acid sequence set forth in at least one of SEQ ID NOs: 2, 3 and 13 to 21; and (b) a protein which consists of an amino acid sequence resulting from deletion, substitution, insertion or addition of one amino acid or a plurality of amino acids in the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function; and (c) a protein which consists of an amino acid sequence having a sequence identity of 80% or higher to the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function. ### Claim 17 (depends on 3) The fusion protein of claim 3, which is one of the following (a) to (c): (a) a protein which consists of an amino acid sequence set forth in at least one of SEQ ID NOs: 2, 3 and 13 to 21; and (b) a protein which consists of an amino acid sequence resulting from deletion, substitution, insertion or addition of one amino acid or a plurality of amino acids in the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function; and (c) a protein which consists of an amino acid sequence having a sequence identity of 80% or higher to the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function. ### Claim 18 (depends on 4) The fusion protein of claim 4, which is one of the following (a) to (c): (a) a protein which consists of an amino acid sequence set forth in at least one of SEQ ID NOs: 2, 3 and 13 to 21; and (b) a protein which consists of an amino acid sequence resulting from deletion, substitution, insertion or addition of one amino acid or a plurality of amino acids in the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function; and (c) a protein which consists of an amino acid sequence having a sequence identity of 80% or higher to the amino acid sequence (a), and which has glucose dehydrogenase activity and electron transfer function.