Engineered pantothenate kinase variant enzymes

Inventors: Margie Tabuga Borra-Garske (Palo Alto, CA); Oscar Alvizo (Fremont, CA); Lillian Jasmine Miller (Saratoga, CA); Jesse B. Slaton (Sacramento, CA); Aksiniya Lyubenova Petkova (Sunnyvale, CA); Nandhitha Subramanian (Cambridge, GB); Joshua Kolev (Califon, NJ); Anna Fryszkowska (New York, NY); Agustina Rodriguez-Granillo (West Newton, MA); Grant Murphy (Princeton, NJ)

Assignee: Codexis, Inc.

C12N9/1205 C12Y207/01033

View Patent ↗

Loading inventors, assignments & file history…

Monitor This Case

Get email alerts when status or documents change.

Order Certified Copies

Most orders are placed with the USPTO same day — all within 24 business hours.

Order via The Patent Place →

Pre-filled with this patent's details

Quick Facts

Patent No.: US 12,110,513
App. No.: 17/120,646
Granted: Oct 8, 2024
Kind: B2

External Links

USPTO Patent Center↗ Full Patent PDF↗

Abstract

The present invention provides engineered pantothenate kinase (PanK) enzymes, polypeptides having PanK activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PanK enzymes are also provided. The present invention further provides compositions comprising the PanK enzymes and methods of using the engineered PanK enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

Claims (17)

1. An engineered pantothenate kinase polypeptide comprising an amino acid sequence having at least 85% sequence to SEQ ID NO: 2, wherein said engineered pantothenate kinase polypeptide comprises an amino acid substitution set corresponding to amino acid positions F15L, V27N, and W283H/L in SEQ ID NO: 2, wherein said engineered pantothenate kinase polypeptide has increased ability to convert ethynyl glyceraldehyde to ethynyl glyceraldehyde phosphate when compared to the wild-type polypeptide having the amino acid sequence of SEQ ID NO: 2.

2. The engineered pantothenate kinase polypeptide of claim 1 , wherein the amino acid substitutions are F15L/V27N/W283L.

3. The engineered pantothenate kinase polypeptide of claim 1 , wherein the amino acid substitutions are F15L/V27N/W283H.

4. The engineered pantothenate kinase polypeptide of claim 3 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions of L277I/I281M.

5. The engineered pantothenate kinase polypeptide of claim 4 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 14/19/41/157/161/261, 19/22/41/44/54/119/157/261/298/308, 19/22/54/157/169, 22/106/218, 41, 41/44/54/119/120/157/169/261, 41/44/54/119/120/157/261/298/308, 41/44/54/119/120/161/169/261/298, 41/44/54/119/298/305, 41/44/161/169/261, 41/44/169/261/298/308, 41/44/169/261/308, 41/54/119/157/169/261, 41/119/161/169/261/308, 41/119/161/308, 44/54/119/120/157/161/169, 44/54/119/120/157/161/261, 44/54/119/120/169/261, 44/54/119/157/161/261/298, 44/54/119/169, 44/76/119/157/161, 44/119/120/261, 44/119/157/161, 44/119/161/261/298, 44/157/161/169, 44/157/298, 44/261/298/308, 44/261/308, 54/119/157/161/169, 54/157/161/261/308, 119/157/161, 119/157/161/169/261, 119/169/261, 119/261/298/308, 120/157/261, 157, 157/161/169/261, 157/161/308, 157/169/261/298/308, 157/308, 250, 302, and 310.

6. The engineered pantothenate kinase polypeptide of claim 5 , wherein the amino acid substitutions are F15L/V27N/154V/V157M/S161K/N261G/L277I/I281M/W283H/A308I.

7. The engineered pantothenate kinase polypeptide of claim 6 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 24/48, 64, 71, 92/301, 123, 125, 134, and 180.

8. The engineered pantothenate kinase polypeptide of 7, wherein the amino acid substitutions are F15L/V27N/154V/L123H/V157M/S161K/N261G/L277I/I281M/W283H/A308I.

9. The engineered pantothenate kinase polypeptide of claim 8 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 83/84/305, 143, and 154.

10. The engineered pantothenate kinase polypeptide of claim 9 , wherein the amino acid substitutions are F15L/V27N/154V/L123H/K143G/V157M/S161K/N261G/L277I/I281M/W83H/A308I.

11. The engineered pantothenate kinase polypeptide of claim 10 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 12/169/213/247/283/288, 16/247, 49/247, 64/104/154/284, 75/104/284, 169/247, and 247.

12. The engineered pantothenate kinase polypeptide of claim 11 , wherein the amino acid substitutions are F15L/V27N/154V/V75Q/T104F/L123H/K143G/V157M/S161K/N261G/L277I/I281M/W283H/L284V/A308I.

13. The engineered pantothenate kinase polypeptide of claim 12 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 8, 8/13/14/83/247, 8/64/213/247, 8/64/213/247, 8/64/247, 10, and 264/276.

14. The engineered pantothenate kinase polypeptide of claim 12 , wherein the engineered pantothenate kinase polypeptide further comprises amino acid substitutions selected from the group consisting of amino acid positions 11/13/23/61/85/304, 13/19/61, 23/61, and 23/61/304.

15. A composition comprising the engineered pantothenate kinase of claim 1 .

16. A polynucleotide sequence encoding the engineered pantothenate kinase of claim 1 .

17. A vector comprising the polynucleotide sequence of claim 16 .

Assignments (1)

SECURITY INTEREST Recorded Feb 15, 2024

From: CODEXIS, INC.

To: INNOVATUS LIFE SCIENCES LENDING FUND I, LP, AS COLLATERAL AGENT

Reel/Frame 066600/0650 →

Continuity (4)

Division 16460147 · Jul 2, 2019

Provisional Application 62695587 · Jul 9, 2018

Provisional Application 62822301 · Mar 22, 2019

Related Publication 20210102179A1 · Apr 8, 2021