Methods for making L-glufosinate

Patent metadata

Field	Value
Publication number	US-11913048-B2
Application number	US-202318179136-A
Country	US
Kind code	B2
Filing date	Mar 6, 2023
Priority date	Mar 2, 2016
Publication date	Feb 27, 2024
Grant date	Feb 27, 2024

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.

First claim

What is claimed is:

1. A method for making L-glufosinate, comprising:
reacting D-glufosinate with a D-amino acid oxidase (DAAO) enzyme to form PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid);
aminating the PPO to L-glufosinate by an L-amino acid dehydrogenase (LAAD) enzyme in the presence of an ammonia source,
wherein at least 70% of the D-glufosinate is converted to L-glufosinate to obtain a composition comprising L-glufosinate, PPO, and D-glufosinate; and
subjecting the composition to a purification method selected from ion exchange, extraction, salt formation, crystallization, filtration, and combinations thereof.

2. 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.

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

4. 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.

5. The method of claim 4 , wherein the mutation at position 54 is selected from the group consisting of N54C, N54L, N54T, and N54V.

6. The method of claim 4 , wherein the mutation at position 56 is T56M.

7. The method of claim 4 , wherein the mutation at position 58 is selected from the group consisting of F58A, F58G, F58H, F58K, F58N, F58Q, F58R, F58S, and F58T.

8. The method of claim 4 , wherein the mutation at position 213 is M213S.

9. The method of claim 4 , wherein the mutant DAAO comprises mutations at positions 54 and 56, using SEQ ID NO: 2 as a reference sequence.

10. The method of claim 4 , wherein the mutant DAAO comprises mutations F580 or F58H, using SEQ ID NO: 2 as a reference sequence.

11. The method of claim 4 , wherein the mutant DAAO comprises mutations N54V and F58Q, using SEQ ID NO: 2 as a reference sequence.

12. The method of claim 4 , wherein the mutant DAAO comprises mutations N54V, F58Q, and M213S, using SEQ ID NO: 2 as a reference sequence.

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

14. The method of claim 13 , wherein all reagents are substantially added at the start of the reaction.

15. The method of claim 13 , wherein the reagents for the reacting step and the reagents for the aminating step are added to the single container at different times.

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

17. The method of claim 1 , wherein the reacting step is conducted while aerating.

18. The method of claim 17 , wherein the aerating is conducted intermittently or continuously.

19. The method of claim 17 , wherein the aerating comprises introducing oxygen, oxygen enriched air, an oxygen enriched gas stream, or air.

Assignees

• Basf Se

Inventors

• Green Brian Michael
• Gradley Michelle Lorraine

Classifications

• C12N9/0014

  acting on the CH-NH2 group of donors (1.4) · CPC title

• C12N9/0024

  D-Amino acid oxidase (1.4.3.3) · CPC title

• C12Y104/03003

  D-Amino-acid oxidase (1.4.3.3) · CPC title

• C12P41/00 Primary

  Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture · CPC title

• C12P13/04 Primary

  Alpha- or beta- amino acids {(other amino acids C12P13/005)} · CPC title