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Biocatalysts for ezetimibe synthesis
US9644189B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9644189-B2 |
| Application number | US-201615179503-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jun 10, 2016 |
| Priority date | May 4, 2010 |
| Publication date | May 9, 2017 |
| Grant date | May 9, 2017 |
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- Title
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- Abstract
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- Assignees and inventors
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Abstract
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The present disclosure relates to non-naturally occurring polypeptides useful for preparing Ezetimibe, polynucleotides encoding the polypeptides, and methods of using the polypeptides.
First claim
Opening claim text (preview).
What is claimed is: 1. A polynucleotide encoding a non-naturally occurring polypeptide having ketoreductase activity comprising an amino acid sequence that has at least 80% sequence identity to SEQ ID NO:2 and at least the following features: residue at position X40 is R; residue at position X153 is I, or L; residue at position X190 is A, or P; residue at position X196 is A, C, N, S, or T; residue at position X199 is F, or W; and residue at position X206 is I. 2. The polynucleotide of claim 1 , wherein the amino acid sequence of said encoded non-naturally occurring ketoreductase polypeptide further comprises a group of features selected from: the residue at position X93 is A and at position X94 is T; the residue at position X93 is A and at position X94 is P; the residue at position X93 is A and at position X94 is S; or the residue at position X93 is I and at position X94 is S. 3. The polynucleotide of claim 1 , wherein the amino acid sequence of said encoded non-naturally occurring ketoreductase polypeptide further comprises a group of features selected from: the residue at position X203 is G; the residue at position X202 is G and at position X203 is G; or the residue at position X201 is A, position X202 is G and at position X203 is G. 4. The polynucleotide of claim 1 , wherein the amino acid sequence of said encoded non-naturally occurring ketoreductase polypeptide comprises: the residue at position X190 is A; the residue at position X196 is T; and the residue at position X199 is F. 5. The polynucleotide of claim 1 , wherein the amino acid sequence of said encoded non-naturally occurring ketoreductase polypeptide further comprises one or more of the following features: 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. 6. An expression vector comprising the polynucleotide of claim 1 . 7. An expression vector comprising the polynucleotide of claim 2 . 8. An expression vector comprising the polynucleotide of claim 3 . 9. An expression vector comprising the polynucleotide of claim 4 . 10. An expression vector comprising the polynucleotide of claim 5 . 11. The expression vector of claim 6 , further comprising at least one control sequence. 12. A host cell comprising the polynucleotide of claim 1 . 13. A host cell comprising the expression vector of claim 6 . 14. A host cell comprising the expression vector of claim 7 . 15. A host cell comprising the expression vector of claim 8 . 16. A host cell comprising the expression vector of claim 9 . 17. A host cell comprising the expression vector of claim 10 . 18. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 12 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell. 19. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 13 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell. 20. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 14 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell. 21. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 15 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell. 22. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 16 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell. 23. A method for preparing a non-naturally occurring ketoreductase polypeptide comprising culturing a host cell of claim 17 , under conditions such that said non-naturally occurring ketoreductase polypeptide is produced and isolating the non-naturally occurring polypeptide from the cell.
Assignees
Inventors
Classifications
Carbonyl reductase (NADPH) (1.1.1.184) · CPC title
Oxidoreductases acting on the CH-OH group of donors (1.1) · CPC title
- C12N9/0006Primary
acting on CH-OH groups as donors (1.1) · CPC title
Nitrogen or oxygen as hetero atom and at least one other diverse hetero ring atom in the same ring · CPC title
from mammals · CPC title
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Frequently asked questions
Answers are generated from the same data shown on this page.
- What does patent US9644189B2 cover?
- The present disclosure relates to non-naturally occurring polypeptides useful for preparing Ezetimibe, polynucleotides encoding the polypeptides, and methods of using the polypeptides.
- Who is the assignee on this patent?
- Codexis Inc, Codexis Inc
- What technology area does this patent fall under?
- Primary CPC classification C12N9/0006. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue May 09 2017 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).