IP Library › Granted Patent US 12,305,207
Granted Patent B2
US 12,305,207 · App. 18/435,544 · Granted May 20, 2025

Methods for making L-glufosinate

Inventors: Brian Michael Green (Lutherville, MD); Michelle Lorraine Gradley (Canterbury, GB)
Assignee: BASF SE
C12P13/02A01N25/30A01N57/20C12N1/20C12N9/10C12P9/00C12P13/04C12P41/00C12Y104/03003C12Y206/01019
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Quick Facts
Patent No.
US 12,305,207
App. No.
18/435,544
Granted
May 20, 2025
Kind
B2
Abstract

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

Claims (22)

1. A method for making L-glufosinate, comprising:

reacting D-glufosinate with a D-amino acid oxidase (DAAO) enzyme to form 2-oxo-4-(hydroxy(methyl)phosphinoyl) butyric acid (PPO); and

aminating the PPO to L-glufosinate using an L-amino acid dehydrogenase (LAAD) enzyme in the presence of an ammonia source and a redox cofactor acceptor.

2. The method according to claim 1 , further comprising the step of recycling the redox cofactor acceptor.

3. The method according to claim 1 , wherein the D-glufosinate is originally present in a racemic mixture of D- and L-glufosinate or salts thereof.

4. The method according to claim 1 , wherein the DAAO enzyme has the amino acid sequence of SEQ ID NO: 2.

5. The method of claim 1 , wherein the DAAO enzyme is a mutant DAAO comprising one or more mutations at positions 54, 56, 58, 213, and 238, using SEQ ID NO: 2 as a reference sequence.

6. The method of claim 5 , wherein the mutation at position 54 is selected from the group consisting of N54C, N54L, N54T, and N54V.

7. The method of claim 5 , wherein the mutation at position 56 is T56M.

8. The method of claim 5 , wherein the mutation at position 58 is selected from the group consisting of F58A, F58G, F58H, F58K, F58N, F58Q, F58R, F58S, and F58T.

9. The method of claim 5 , wherein the mutation at position 213 is M213S.

10. The method of claim 5 , wherein the mutant DAAO comprises mutations at positions 54 and 56, using SEQ ID NO: 2 as a reference sequence.

11. The method of claim 5 , wherein the mutant DAAO comprises mutations F58Q or F58H, using SEQ ID NO: 2 as a reference sequence.

12. The method of claim 5 , wherein the mutant DAAO comprises mutations N54V and F58Q, using SEQ ID NO: 2 as a reference sequence.

13. The method of claim 5 , wherein the mutant DAAO comprises mutations N54V, F58Q, and M213S, using SEQ ID NO: 2 as a reference sequence.

14. The method of claim 1 , wherein the reacting step and the aminating step are performed in a single container.

15. The method of claim 14 , wherein all reagents are substantially added at the start of the reaction.

16. The method of claim 14 , wherein the reagents for the reacting step and the reagents for the aminating step are added to the single container at different times.

17. The method of claim 1 , wherein the reacting step and the aminating step are performed in separate containers.

18. The method of claim 1 , wherein reacting D-glufosinate with the DAAO enzyme is conducted while aerating.

19. The method of claim 18 , wherein the aerating comprises introducing oxygen, oxygen enriched air, an oxygen enriched gas stream, or air.

20. The method of claim 18 , wherein the aerating is conducted intermittently or continuously.

Continuity (10)
Continuation 18179136 · Mar 6, 2023
Continuation 17530018 · Nov 18, 2021
Continuation 16997133 · Aug 19, 2020
Continuation 16287290 · Feb 27, 2019
Continuation 15787448 · Oct 18, 2017
Division 15445254 · Feb 28, 2017
Provisional Application 62413240 · Oct 26, 2016
Provisional Application 62336989 · May 16, 2016
Provisional Application 62302421 · Mar 2, 2016
Related Publication 20240301452A1 · Sep 12, 2024
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