# US 12,110,512 B2 · Biocatalysts for ezetimibe synthesis > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,110,512 B2 - Document ID: US12110512B2 - Status: Granted - Kind: B2 - Application number: 17/204,189 - Filed: 2021-03-17 - Published: 2021-07-22 - Granted: 2024-10-08 - Earliest claimed: 2010-05-04 - Assignees: Codexis, Inc. - Applicants: Codexis Inc. - Inventors: Michael A. Crowe; Oscar Alvizo; Behnaz Behrouzian; Yong Koy Bong; Steven J. Collier; Anupam Gohel; Jagadeesh Mavinahalli; Naga K. Modukuru; Emily Mundorff; Derek J. Smith; Shiwei Song; Wan Lin Yeo - Web page: https://patentgrep.com/patent/US12110512B2 ## CPC - C12N 9/0006: {acting on CH-OH groups as donors (1.1)} - C07K 14/47: from mammals - C12P 17/10: Nitrogen as only ring hetero atom - C12Y 101/00: Oxidoreductases acting on the CH-OH group of donors (1.1) - C12Y 101/01184: Carbonyl reductase (NADPH) (1.1.1.184) - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - C12P 17/14: Nitrogen or oxygen as hetero atom and at least one other diverse hetero ring atom in the same ring - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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)} - C12Y: ENZYMES - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract The present disclosure relates to non-naturally occurring polypeptides useful for preparing Ezetimibe, polynucleotides encoding the polypeptides, and methods of using the polypeptides. ## Claims ### Claim 1 A non-naturally occurring ketoreductase polypeptide having ketoreductase activity, comprising an amino acid sequence that has at least 95% sequence identity to SEQ ID NO: 168, wherein the residue at position 40 is arginine. ### Claim 2 (depends on 1) The non-naturally occurring ketoreductase polypeptide of claim 1, wherein the amino acid sequence of said non-naturally occurring ketoreductase polypeptide further comprises the following feature: the residue at position 146 is glycine. ### Claim 3 (depends on 1) The non-naturally occurring ketoreductase polypeptide of claim 1, wherein the amino acid sequence of said non-naturally occurring ketoreductase polypeptide further comprises one or more of the features selected from: the residue at position X93 is A and the residue at position X94 is T; the residue at position X93 is A and the residue at position X94 is P; the residue at position X93 is A and the residue at position X94 is S; or the residue at position X93 is I and the residue at position X94 is S. ### Claim 4 (depends on 1) The non-naturally occurring ketoreductase of claim 1, wherein the amino acid sequence of said non-naturally occurring ketoreductase polypeptide further comprises one or more of the features selected from: the residue at position X203 is G; the residue at position X202 is G and the residue at position X203 is G; or the residue at position X201 is A, the residue position X202 is G and the residue at position X203 is G. ### Claim 5 (depends on 1) The non-naturally occurring ketoreductase claim 1, wherein the amino acid sequence of said non-naturally occurring ketoreductase polypeptide further comprises one or more of the features selected from: the residue at position X196 is T; and the residue at position X199 is F. ### Claim 6 (depends on 1) The non-naturally occurring ketoreductase of claim 1, wherein the amino acid sequence of said non-naturally occurring ketoreductase polypeptide further comprises one or more of the features selected from: the residue at position X21 is R or F; the residue at position X25 is R, T, or N; the residue at position X64 is V; the residue at position X93 is A; the residue at position X94 is T, S, or P; the residue at position X95 is M, or V; the residue at position X96 is V, G, A, N, S, P, or T; the residue at position X99 is L; the residue at position X108 is H or K; the residue at position X117 is G; the residue at position X127 is K, or Q; the residue at position X147 is M, or I; the residue at position X148 is I; the residue at position X150 is H; the residue at position X152 is F, or N; the residue at position X155 is C; the residue at position X163 is I; the residue at position X195 is M; the residue at position X201 is I, L, or A; the residue at position X202 is L, N, V, or G; the residue at position X203 is G; the residue at position X204 is V, or A; the residue at position X205 is V; the residue at position X206 is I; the residue at position X207 is T, C, I, or N; the residue at position X211 is K; the residue at position X221 is D; the residue at position X223 is I; or the residue at position X226 is V.