Biocatalytic synthesis of quinic acid and conversion to hydroquinone

• US 20060003429A1
• Filed: 07/19/2005
• Published: 01/05/2006
• Est. Priority Date: 01/29/1999
• Status: Active Grant

• Current Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Original Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Inventors: Frost, John W., Frost, Karen M.
• Granted Patent: US 7,642,083 B2
• Time in Patent Office: Days
• US Class Current: 435/156
• CPC Class Codes: C07C 39/08 Dihydroxy benzenes; Alkylated derivatives thereof
• CPC Class Codes: C12P 7/22 aromatic
• CPC Class Codes: C12P 7/26 Ketones
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids

Abstract

A bioengineered synthesis scheme for the production of quinic acid from a carbon source is provided. Methods of producing quinic acid from a carbon source based on the synthesis scheme as well as conversion of quinic acid to hydroquinone are also provided.

Claims

1. A recombinant microbe comprising:

(A) a 3-deoxy-D-arabino-heptulosonic acid-7-phosphate (DAHP) synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids,

(B) a 3-dehydroquinate (DHQ) synthase enzyme, and

(C) a shikimate dehydrogenase enzyme,

wherein

(I) at least one of said enzymes is encoded by recombinant DNA therein,

(II) the recombinant microbe does not cause dehydration of 3-dehydroquinate to dehydroshikimate, and

(III) the recombinant microbe is capable of producing quinic acid from a carbon source by a synthesis scheme that uses all three of said enzymes.

2. The recombinant microbe according to claim 1, wherein the DAHP synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids is encoded by recombinant DNA in said microbe.

3. The recombinant microbe according to claim 1, wherein the DHQ synthase enzyme is encoded by recombinant DNA in said microbe.

4. The recombinant microbe according to claim 1, wherein the shikimate dehydrogenase enzyme is encoded by recombinant DNA in said microbe.

5. The recombinant microbe according to claim 1, wherein the microbe'"'"'s DNA that encodes dehydroquinate dehydratase has been inactivated so that the recombinant microbe does not cause dehydration of 3-dehydroquinate to dehydroshikimate.

6. The recombinant microbe according to claim 1, wherein the microbe is selected from the group consisting of fungi, yeasts, and bacteria.

7. The recombinant microbe according to claim 1, wherein the DAHP synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids is encoded by an aroF^FBR gene.

8. The recombinant microbe according to claim 1, wherein the DHQ synthase enzyme is encoded by an aroB gene.

9. The recombinant microbe according to claim 1, wherein the shikimate dehydrogenase enzyme is encoded by an aroE gene.

10. The recombinant microbe according to claim 9, wherein the aroE gene is an E. coli aroE gene.

11. The recombinant microbe according to claim 10, wherein the aroE gene is comprised by a plasmid.

12. The recombinant microbe according to claim 11, wherein the plasmid is pKD12.112.

13. The recombinant microbe according to claim 1, wherein the DAHP synthase, DHQ synthase, and shikimate dehydrogenase are respectively encoded by E. coli aroF^FBR, E. coli aroB, and E. coli aroE genes.

14. The recombinant microbe according to claim 1, wherein the recombinant microbe is a recombinant E. coli.

15. The recombinant microbe according to claim 5, wherein the microbe'"'"'s inactivated DNA that encodes dehydroquinate dehydratase comprises an inactivated aroD gene.

16. The recombinant microbe according to claim 15, wherein the recombinant microbe is E. coli QP1.1.

17. A bioengineered synthesis scheme for quinic acid production comprising

(A) a 3-deoxy-D-arabino-heptulosonic acid-7-phosphate (DAHP) synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids,

(B) a 3-dehydroquinate (DHQ) synthase enzyme, and

(C) a shikimate dehydrogenase enzyme,

wherein at least one of said enzymes is a recombinant enzyme,

the synthesis scheme involving

(I) synthesis of DAHP from erythrose-4-phosphate (E4P) and phosphoenolpyruvate (PEP) by the DAHP synthase,

(II) conversion of DAHP to DHQ by the DHQ synthase, and

(III) conversion of DHQ to quinic acid by the shikimate dehydrogenase.

18. The scheme according to claim 17, wherein the synthesis scheme is present where dehydration of DHQ to dehydroshikimate does not occur.

19. The synthesis scheme according to claim 18, wherein said scheme is present in a recombinant microbe that does not cause dehydration of DHQ to dehydroshikimate.

20. The synthesis scheme according to claim 19, wherein the recombinant microbe is selected from the group consisting of fungi, yeasts, and bacteria.

21. A method for the production of quinic acid comprising

(A) providing a synthesis scheme according to claim 17, erythrose-4-phosphate (E4P), and phosphoenolpyruvate (PEP);

and

(B) contacting the synthesis scheme with the E4P and PEP under conditions in which the enzymes of said synthesis scheme can catalyze the formation of quinic acid therefrom;

thereby producing quinic acid.

22. The method according to claim 21, wherein

(i) the synthesis scheme is provided by a recombinant microbe comprising

(A) a 3-deoxy-D-arabino-heptulosonic acid-7-phosphate (DAHP) synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids,

(B) a 3-dehydroquinate (DHQ) synthase enzyme, and

(C) a shikimate dehydrogenase enzyme,

(I) at least one of said enzymes being encoded by recombinant DNA therein,

(II) the recombinant microbe not causing dehydration of 3-dehydroquinate to dehydroshikimate, and

(III) the recombinant microbe being capable of producing quinic acid from a carbon source by a synthesis scheme that uses all three of said enzymes;

and

(ii) the E4P and PEP are provided by the recombinant microbe using a carbon source.

23. A method for the production of hydroquinone from quinic acid, comprising

(A) providing quinic acid prepared by operation of a synthesis scheme according to claim 17, and an oxidizing agent;

(B) contacting the quinic acid with the oxidizing agent to produce 3,4,5-trihydroxycyclohexanone; and

(C) heating the 3,4,5-trihydroxycyclohexanone to produce hydroquinone.

24. The method according to claim 23, wherein the oxidizing agent is selected from the group consisting of HOCl, AgNO₃, (NH₄)₂Ce(NO₃)₆, V₂O₅, NH₄VO₃, and 2 KHSO₅.KHSO₄.K₂SO₄/NaCl.

25. The method according to claim 23, wherein the synthesis scheme is provided by a recombinant microbe comprising

(A) a 3-deoxy-D-arabino-heptulosonic acid-7-phosphate (DAHP) synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids,

(B) a 3-dehydroquinate (DHQ) synthase enzyme, and

(C) a shikimate dehydrogenase enzyme,

(I) at least one of said enzymes being encoded by recombinant DNA therein,

(II) the recombinant microbe not causing dehydration of 3-dehydroquinate to dehydroshikimate, and

the recombinant microbe being capable of producing quinic acid from a carbon source by a synthesis scheme that uses all three of said enzymes.