Methods for making L-glufosinate

Patent metadata

Field	Value
Publication number	US-11560577-B2
Application number	US-202016997133-A
Country	US
Kind code	B2
Filing date	Aug 19, 2020
Priority date	Mar 2, 2016
Publication date	Jan 24, 2023
Grant date	Jan 24, 2023

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) while aerating; and
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; and
wherein the method is performed ex vivo.

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 F58Q 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 aerating comprises introducing oxygen, oxygen enriched air, an oxygen enriched gas stream, or air.

18. The method of claim 1 , wherein the aerating is conducted intermittently or continuously.

Assignees

• Basf Se

Inventors

• Green Brian Michael
• Gradley Michelle Lorraine

Classifications

• C12P13/02 Primary

  Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)} · CPC title

• C12P9/00

  Preparation of organic compounds containing a metal or atom other than H, N, C, O, S or halogen {(phosphoglycerides, C12P7/6481)} · CPC title

• C12P41/00 Primary

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

• A01N57/20

  containing acyclic or cycloaliphatic radicals · CPC title

• A01N25/30

  characterised by the surfactants · CPC title