Methods for making L-glufosinate
AssigneeAgriMetis, LLC
InventorsBrian Michael Green, Michelle Lorraine Gradley, Annette Alcasabas
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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.
Claims18
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 a transaminase (TA) enzyme, using an amine group from one or more amine donors,
- 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 further purification methods selected from ion exchange, extraction, salt formation, crystallization, and filtration.
A D-amino acid oxidase (DAAO) enzyme comprising the amino acid sequence set forth in SEQ ID NO:2, wherein said enzyme comprises one or more mutations at positions 54, 56, 58, 213, and 238;
- wherein said mutation at position 54 is selected from the group consisting of N54C, N54L, N54T, and N54V;
- said mutation at position 56 is T56M;
- said mutation at position 58 is selected from the group consisting of F58A, F58G, F58H, F58K, F58N, F58Q, F58R, F58S, and F58T;
- and said mutation at position 213 is M213S.
Dependencies are inferred from the claim text.