Method for the preparation of chiral alpha haloalkanoic acids

Patent metadata

Field	Value
Publication number	US-11053518-B2
Application number	US-201816650311-A
Country	US
Kind code	B2
Filing date	Sep 24, 2018
Priority date	Sep 28, 2017
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

What is described herein relates to a method of selectively hydrolyzing an enantiomer of an alpha haloalkanoic acid according to formula I employing a polypeptide having dehalogenase activity comprising an amino acid sequence as set forth in SEQ ID NO. 1 or SEQ ID NO. 4 or a sequence with at least 80% sequence identity to either of said sequences and to the use of said method.

First claim

The invention claimed is:

1. A method of selectively hydrolyzing the S-enantiomer of an alpha haloalkanoic acid according to formula I,

wherein X is bromide or chloride and
R is an alkyl chain of 1 to 6 carbon atoms, wherein said alkyl chain is straight or branched at carbon atoms γ or δ,
comprising:
providing a racemate of the R-enantiomer and the S-enantiomer of said alpha haloalkanoic acid,
providing a polypeptide having dehalogenase activity comprising the amino acid sequence as set forth in SEQ ID NO. 1,
reacting the racemate for 1-8 hours, wherein
the pH is in the range of 9-10 and the temperature is in the range of 15-35° C. for the polypeptide with dehalogenase activity comprising the amino acid sequence as set forth in SEQ ID NO. 1,
and wherein an enantiomeric excess of the R-enantiomer of the haloalkanoic acid of between 90.0% and 99.9% is reached after 1-8 hours, and wherein the concentration of the racemate of said alpha haloalkanoic acid according to Formula I is between 80 and 200 g/L and the polypeptide with dehalogenase activity is comprised within whole cells.

2. The method according to claim 1 , wherein the ratio of racemate of the alpha haloalkanoic acid according to Formula I to biomass of whole cells comprising the polypeptide having dehalogenase activity comprising the amino acid sequence as set forth in SEQ ID NO. 1 or or a sequence with at least 90% sequence identity to said sequence is between 2:1 to 15:1.

3. The method according to claim 1 , wherein moiety R of said alpha haloalkanoic acid of formula I is chosen from the group consisting of ethyl, butyl, 2-methyl-propyl and methyl-cyclopropyl.

4. The method according to claim 1 for selective hydrolysis of the S-enantiomer of an alpha haloalkanoic acid of formula I from a racemate, wherein the enantiomeric excess of the remaining R-enantiomer of said alpha haloalkanoic acid is between 90.0% and 99.9%.

5. A method of selectively hydrolyzing the S-enantiomer of an alpha haloalkanoic acid according to formula II,

wherein
R is an alkyl chain of 1 to 6 carbon atoms, wherein said alkyl chain is straight or branched at carbon atoms γ or δ, and F is a single fluorine atom,
said method comprising:
providing a racemate of the R-enantiomer and the S-enantiomer of said alpha haloalkanoic acid,
providing a polypeptide having dehalogenase activity comprising the amino acid sequence as set forth in SEQ ID NO. 4,
reacting the racemate for 1-8 hours, wherein the pH is in the range of 9-10 and the temperature is in the range of 55-65° C. for the polypeptide with dehalogenase activity comprising the amino acid sequence as set forth in SEQ ID NO. 4,
wherein an enantiomeric excess of the R-enantiomer of the haloalkanoic acid of between 90.0% and 99.9% is reached after 1-8 hours, and
wherein the concentration of the racemate of said alpha haloalkanoic acid according to Formula II is between 80 and 200 g/L and the polypeptide with dehalogenase activity is comprised within whole cells.

6. The method according to claim 2 , wherein the ratio is between 3:1 to 10:1.

7. The method according to claim 2 , wherein the ratio is 4:1.

8. The method according to claim 1 , wherein X is bromide.

9. The method according to claim 1 , wherein X is chloride.

10. The method according to claim 3 , wherein moiety R of said alpha haloalkanoic acid of formula I is ethyl.

11. The method according to claim 3 , wherein moiety R of said alpha haloalkanoic acid of formula I is butyl.

12. The method according to claim 3 , wherein moiety R of said alpha haloalkanoic acid of formula I is 2-methyl-propyl.

13. The method according to claim 3 , wherein moiety R of said alpha haloalkanoic acid of formula I is methyl-cyclopropyl.

Assignees

• Bayer Ag

Inventors

• Spelberg Markus
• Egger Julian

Classifications

• C12P41/00 Primary

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

• C12P7/40 Primary

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• C12P41/002

  by oxidation/reduction reactions · CPC title

• C12Y308/01002

  (S)-2-Haloacid dehalogenase (3.8.1.2) · CPC title