# US 11,297,862 B2 · Steviol glycoside transport > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,297,862 B2 - Document ID: US11297862B2 - Status: Granted - Kind: B2 - Application number: 15/751,674 - Filed: 2016-08-15 - Published: 2018-08-23 - Granted: 2022-04-12 - Earliest claimed: 2015-08-13 - Assignees: Dsm IP Assets B.V. - Applicants: Dsm IP Assets B.V. - Inventors: Viktor Marius Boer; Priscilla Zwartjens; Eric Van Den Berg - Web page: https://patentgrep.com/patent/US11297862B2 ## CPC - A23L 27/36: {Terpene glycosides} - A23L 2/60: Sweeteners - C07K 14/39: from yeasts - C07K 14/395: from Saccharomyces - C07K 14/40: from Candida - C12N 1/16: Yeasts; Culture media therefor - C12N 9/2402: {hydrolysing O- and S- glycosyl compounds (3.2.1)} - C12P 15/00: Preparation of compounds containing at least three condensed carbocyclic rings {(gibbanes C12P27/00; naphthacenes C12P29/00)} - C12P 19/56: having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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)} ## Abstract A recombinant host capable of producing a steviol glycoside which overexpresses a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto. A recombinant host capable of producing a steviol glycoside which has been modified, preferably in its genome, to result in a deficiency in the production of a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto. ## Claims ### Claim 1 A recombinant cell capable of producing a steviol glycoside which overexpresses a nucleic acid encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:29 or an amino acid sequence having at least 98% sequence identity to the amino acid sequence set forth in SEQ ID NO:29. ### Claim 2 (depends on 1) The recombinant cell according to claim 1, which further comprises one or more recombinant nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity. ### Claim 3 (depends on 1) The recombinant cell according to claim 1, wherein the host belongs to one of the genera Saccharomyces, Aspergillus, Pichia, Kluyveromyces, Candida, Hansenula, Humicola, Issatchenkia, Trichosporon, Brettanomyces, Pachysolen, Yarrowia, Yamadazyma, or Escherichia. ### Claim 4 (depends on 3) The recombinant host according to claim 3, wherein the recombinant host is a Saccharomyces cerevisiae cell, a Yarrowia lipolytica cell, a Candida krusei cell, or an Escherichia coli cell. ### Claim 5 (depends on 1) The recombinant cell according to claim 1, wherein the polypeptide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO:29. ### Claim 6 (depends on 1) The recombinant cell according to claim 1, wherein the polypeptide mediates steviol glycoside transport and wherein the amount of total produced steviol glycoside outside the cell, as compared to inside the cell, is higher as compared to a corresponding cell which does not overexpress the nucleic acid. ### Claim 7 (depends on 1) A process for the preparation of a steviol glycoside which comprises fermenting the recombinant cell according to claim 1 in a suitable fermentation medium and, optionally, recovering the steviol glycoside. ### Claim 8 (depends on 7) The process according to claim 7 for the preparation of a steviol glycoside, wherein the process is carried out on an industrial scale.