Enzymatic method for producing L-glufosinate and its phosphoesters

1. An enzymatically catalyzed method for producing L-glufosinate or a phosphoester thereof, the method comprising:

reacting an activated L-homoserine H A with a substrate S of the following structure (I) to produce a compound of the following structure (III),

wherein R 1 is selected from the group consisting of hydrogen, an alkyl, an alkenyl, an alkinyl, a hydroxyalkyl, and an aryl, and

wherein the activated L-homoserine HA has the following structure (II):

wherein R 2 is a hydrocarbon group with 1 to 15 carbon atoms which optionally comprises at least one functional group selected from OH, COOH, NH, and

wherein the reaction is catalyzed by at least one enzyme selected from the group consisting of a sulfhydrylase and a cystathionine γ-synthase,

wherein the sulfhydrylase comprises a polypeptide sequence selected from the group consisting of SEQ ID NO: 2 and variants of SEQ ID NO: 2, SEQ ID NO: 6 and variants of SEQ ID NO: 6, SEQ ID NO: 19 and variants of SEQ ID NO: 19, SEQ ID NO: 22 and variants of SEQ ID NO: 22, SEQ ID NO: 23 and variants of SEQ ID NO: 23, SEQ ID NO: 10 and variants of SEQ ID NO: 10, SEQ ID NO: 14 and variants of SEQ ID NO: 14, SEQ ID NO: 18 and variants of SEQ ID NO: 18, SEQ ID NO: 20 and variants of SEQ ID NO: 20, SEQ ID NO: 24 and variants of SEQ ID NO: 24, SEQ ID NO: 25 and variants of SEQ ID NO: 25, SEQ ID NO: 26 and variants of SEQ ID NO: 26, SEQ ID NO: 27 and variants of SEQ ID NO: 27, SEQ ID NO: 28 and variants of SEQ ID NO: 28, and SEQ ID NO: 34 and variants of SEQ ID NO: 34,

wherein the variants of SEQ ID NOs: 2, 6, 10, 14, 18-20, 22-28, and 34 are defined as polypeptide sequences with at least 95% sequence identity to SEQ ID NOs: 2, 6, 10, 14, 18-20, 22-28, and 34 respectively, and

wherein the cystathionine y-synthase comprises a polypeptide sequence selected from the group consisting of SEQ ID NO: 17 and variants of SEQ ID NO: 17. SEQ ID NO: 21 and variants of SEQ ID NO: 21, SEQ ID NO: 29 and variants of SEQ ID NO: 29, SEQ ID NO: 30 and variants of SEQ ID NO: 30, SEQ ID NO: 31 and variants of SEQ ID NO: 31, SEQ ID NO: 32 and variants of SEQ ID NO: 32, and SEQ ID NO: 33 and variants of SEQ ID NO: 33,

wherein the variants of SEQ ID NOs: 17, 21, and 29-33 are defined as polypeptide sequences with at least 95% sequence identity to SEQ ID NOs: 17, 21, and 29-33 respectively.

2. The method according to claim 1 , wherein the activated L-homoserine H A is selected from the group consisting of O-succinyl-L-homoserine and O-acetyl-L-homoserine.

3. The method according to claim 2 , wherein the activated L-homoserine H A is O-acetyl-L-homoserine.

4. The method according to claim 1 , wherein R 1 is selected from the group consisting of hydrogen and an alkyl group.

5. The method according to claim 4 , wherein the alkyl group is selected from the group consisting of methyl, ethyl, and n-butyl.

6. The method according to claim 1 , wherein R 1 is selected from the group consisting of an alkyl, an alkenyl, an alkinyl, a hydroxyalkyl, and an aryl, further comprising saponifying the compound of the structure (III) obtained in the reaction to give L-glufosinate.

7. The method according to claim 1 , wherein the reaction is catalyzed by the sulfhydrylase.

8. The method according to claim 7 , wherein the sulfhydrylase is an O-acetyl homoserine sulfhydrylase or an O-succinyl homoserine sulfhydrylase.

9. The method according to claim 1 , wherein the activated L-homoserine H A is prepared by fermentation of a strain producing activated L-homoserine H A .

10. The method according to claim 9 , wherein the strain producing activated L-homoserine H A is selected from the group consisting of Escherichia sp., Erwinia sp., Serratia sp., Providencia sp., Corynebacterium sp., Pseudomonas sp., Leptospira sp., Salmonella sp., Brevibacterium sp., Hypomononas sp., Chromobacterium sp., and Norcardia sp., fungi.

11. The method according to claim 1 , wherein R 2 is a hydrocarbon group with 1 to 15 carbon atoms comprising at least one functional group selected from the group consisting of OH, COOH, and NH.