Polynucleotides encoding biocatalysts and methods for hydroxylation of chemical compounds
US-10844358-B2 · Nov 24, 2020 · US
Vectors for expression of biocatalysts
US11634695B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11634695-B2 |
| Application number | US-202017072910-A |
| Country | US |
| Kind code | B2 |
| Filing date | Oct 16, 2020 |
| Priority date | Jun 9, 2016 |
| Publication date | Apr 25, 2023 |
| Grant date | Apr 25, 2023 |
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Abstract
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The present invention provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents.
First claim
Opening claim text (preview).
What is claimed is: 1. An expression vector comprising a polynucleotide comprising SEQ ID NO:1007, 1008, or 1009 with up to 20 nucleotide substitutions, insertions, or deletions in the polynucleotide sequence of SEQ ID NO: 1007, 1008, or 1009. 2. The expression vector of claim 1 , additionally comprising an engineered polynucleotide encoding an engineered polypeptide, wherein said engineered polypeptide is capable of converting (S)-pipecolic acid to (2S,5S)-5-hydroxypipecolic acid with greater than 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% diastereomeric excess of (2S,5R)-5-hydroxypipecolic acid. 3. The expression vector of claim 1 , further comprising at least one control sequence. 4. The expression vector of claim 3 , wherein said at least one control sequence comprises a ribosome binding site. 5. The expression vector of claim 3 , wherein said at least one control sequence comprises an Escherichia coli lac operon. 6. The expression vector of claim 1 , further comprises a chloramphenicol resistance gene. 7. The expression vector of claim 2 , wherein said engineered polynucleotide encoding an engineered polypeptide comprises a nucleic acid sequence optimized for expression in Escherichia coli. 8. A host cell comprising the expression vector of claim 1 . 9. A host cell comprising the expression vector of claim 2 . 10. The host cell of claim 8 , wherein the host cell is Escherichia coli. 11. The host cell of claim 9 , wherein the host cell is Escherichia coli. 12. A method of preparing an engineered polypeptide, comprising culturing the host cell of claim 9 , under conditions suitable for expression of the polypeptide. 13. The method of claim 12 , further comprising a step of isolating the engineered polypeptide. 14. A method of preparing an engineered polypeptide, comprising culturing the host cell of claim 10 , under conditions suitable for expression of the polypeptide. 15. The method of claim 14 , further comprising a step of isolating the engineered polypeptide.
Assignees
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Classifications
containing a six-membered hetero ring · CPC title
Procollagen-proline dioxygenase (1.14.11.2), i.e. proline-hydroxylase · CPC title
- C12N9/0071Primary
acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title
with 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors (1.14.11) · CPC title
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- What does patent US11634695B2 cover?
- The present invention provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents.
- Who is the assignee on this patent?
- Codexis Inc
- What technology area does this patent fall under?
- Primary CPC classification C12N9/0071. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Apr 25 2023 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 5 related publications on this page (citations in our corpus or others sharing the same primary CPC).