Yeast-based process for production of l-pac

• US 6,271,008 B1
• Filed: 02/20/2001
• Issued: 08/07/2001
• Est. Priority Date: 06/03/1998
• Status: Expired due to Fees

• Current Assignee: Victoria University
• Original Assignee: Polychip Pharmaceuticals Pty., Ltd. (Opthea Ltd.), Victoria University Of Technology
• Inventors: Trewhella, Maurice Arthur, Smallridge, Andrew John, Del Guidice, Margaret Mary
• Primary Examiner(s): Lilling, Herbert J.
• Application Number:
  US09/701,704
• Time in Patent Office: 168 Days
• Field of Search: 435/148, 435/150, 435/156, 435/942
• US Class Current: 435/148
• CPC Class Codes: C12P 7/26 Ketones
• CPC Class Codes: Y10S 435/942 Saccharomyces cerevisiae

Abstract

This invention relates to yeast-mediated catalysis in organic solvents, and in particular the yeast-mediated condensation between pyruvate and a substituted aromatic aldehyde to yield the corresponding acyloin (hydroxy ketone) compound. The invention provides a method of synthesis of a substituted carbinol compound, comprising the step of subjecting the corresponding substituted aromatic aldehyde to acyloin condensation mediated by a yeast in an organic solvent under non-fermenting conditions. Preferably the yeast is Saccharomyces cerevisiae. In a preferred embodiment, the reaction is that between pyruvate and benzaldehyde to yield phenylacetylcarbinol, the precursor to ephedrine, to yield a product of high enantiomeric purity.

Claims

1. A method of synthesis of a carbinol compound, comprising the step of subjecting the corresponding aromatic aldehyde to an acyloin condensation reaction mediated by a yeast in an organic solvent under non-fermenting conditions, in the presence of an aliphatic alcohol or aliphatic aldehyde, in which the reaction mixture comprises water sufficient to activate the yeast, but not sufficient to form a two phase system.

2. A method according to claim 1, in which the yeast is Saccharomyces cerevisiae.

3. A method according to claim 1, in which the yeast is of a strain specifically adapted to an organic solvent environment or for enhanced reduction efficiency.

4. A method according to claim 1, in which the yeast is active dried yeast.

5. A method according claim 4, in which the aliphatic alcohol or aliphatic aldehyde is ethanol or acetaldehyde respectively.

6. A method according to claim 5, in which the proportion of ethanol or acetaldehyde is 0.1 ml/g yeast.

7. A method according to claim 5, in which the aliphatic alcohol or aliphatic aldehyde is ethanol.

8. A method according to claim 1, in which the reaction is performed at ambient temperature.

9. A method according to claim 1, in which the reacticn is performed at 16-24°

C.

10. A method according to claim 9, in which the reaction is performed at 20°

C.

11. A method according to claim 1, in which the reaction is performed at 0-10°

C.

12. A method according to claim 1, wherein the organic solvent selected from the group consisting of petroleum ether, carbon tetrachloride, hexane, other hydrocarbons, diethyl ether, toluene, and benzene.

13. A method according to claim 1, wherein the organic solvent is petroleum ether.

14. A method according to claim 1, wherein the organic solvent is a mixed organic solvent system.

15. A method according to claim 14, in which the mixed organic solvent system is selected from the group consisting of 2-ethoxyethanol/diethyl ether, pyridine/carbon tetrachloride, chloroform/petroleum ether and ethyl acetate/toluene.

16. A method according to claim 15, in which the mixed organic solvent system comprises 1 to 30% v/v of a polar organic solvent added to a non-polar organic solvent.

17. A method according to claim 16, in which the mixed organic solvent system comprises 1 to 5% v/v of polar organic solvent added to a non-polar organic solvent.

18. A method according to claim 17, in which the polar organic solvent is selected from the group consisting of chloroform, dichloromethane, methyl ethyl ketone, and methyl isobutyl ketone.

19. A method according to claim 18, in which the reaction is performed in chloroform or dichloromethane in petroleum ether.

20. A method according to claim 1, in which the reaction mixture comprises water at a ratio of 0.6 to 1.2 ml water/g of yeast.

21. A method according to claim 1, in which the reaction is conversion of an optionally-substituted aromatic aldehyde to the corresponding acyloin compound by condensation with an optionally-substituted pyruvat compound.

22. A method according to claim 21, in which the aromatic aldehyde substituted with one or more groups selected from the group consisting of alkyl, aryl, halo, nitro, hydroxy, alkoxy, amino, carbonyl, thioxy or thioalkoxy substituted with alkyl, aryl, halo, nitro, hydroxy, alkoxy, amino, carbonyl, thioxy or thioalkoxy alkyl, aryl, halo, nitro, hydroxy, alkoxy, amino, carbonyl, thioxy and thioalkoxy.

23. A method according to claim 21, in which the reaction is conversion of benzaldehyde to phenylcetylcarbinol.

24. A method according to claim 21, in which the pyruvate compound is pyruvic acid.

25. A method according to claim 24, in which the reaction is performed at a pH of between 5 and 6.

26. A method according to claim 21, in which the pyruvate compound is sodium pyruvate.

27. A method according to claim 26, in which the reaction is performed at a pH of between 5 and 8.

28. A method according to claim 1, in which the reaction time is 12 to 24 hours.

29. A method of synthesis of a substituted carbinol compound, comprising the step of subjecting the corresponding substituted aromatic aldehyde to acyloin condensation mediated by a yeast in an organic solvent under non-fermenting conditions, in which the reaction mixture comprises water sufficient to activate the yeast but not sufficient to form a two phase system.

30. A method of synthesis of a substituted carbinol compound, comprising the step of subjecting the corresponding substituted aromatic aldehyde to acyloin condensation mediated by a yeast in an organic solvent under non-fermenting conditions, in which the reaction is performed at 0-10°

C.