Biocatalystic synthesis of quinic acid and conversion to hydroquinone by recombinant microbes

1. A method for production of quinic acid comprising:

(A) providing one or more enzymes selected from:

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

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

(iii) a shikimate dehydrogenase enzyme; wherein at least one of said enzymes is a recombinant enzyme, wherein the DAHP synthase, DHQ synthase, and shikimate dehydrogenase are encoded by E. coli aroF FBR , E. coli aroB, and E. coli aroE genes, respectively; and

(B) allowing the one or more enzymes to catalyze one or more reactions to synthesize quinic acid from erythrose-4-phosphate (E4P) and phosphoenolpyruvate (PEP); thereby producing quinic acid.

2. The method according to claim 1 , wherein the quinic acid is produced by a recombinant microbe, wherein

(a) the one or more enzymes are produced by the recombinant microbe,

(b) at least one of the enzymes are encoded by recombinant DNA,

(c) the recombinant microbe comprises an inactivating mutation in an endogenous DNA encoding a dehydroquinate dehydratase,

(d) the recombinant microbe is capable of producing quinic acid from a carbon source by using all three enzymes, and

(e) the E4P and PEP are produced by the recombinant microbe,

wherein the recombinant microbe is cultured to allow the one or more enzymes to catalyze the one or more reactions to synthesize quinic acid from the E4P and PEP.

3. A method for production of hydroquinone from quinic acid, comprising:

(A) providing quinic acid produced by the method according to claim 1 , 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.

4. The method according to claim 3 , wherein the oxidizing agent is selected from the group consisting of HOCl, AgNO 3 , (NH 4 ) 2 Ce(NO 3 ) 6 , V 2 O 5 , NH 4 VO 3 , and 2KHSO 5 .KHSO 4 .K 2 SO 4 /NaCl.

5. The method according to claim 3 , wherein the quinic acid is produced by a recombinant microbe comprising:

(A) the DAHP synthase enzyme that is insensitive to feedback inhibition by aromatic amino acids,

(B) the DHQ synthase enzyme, and

(C) the shikimate dehydrogenase enzyme, wherein the DAHP synthase, DHQ synthase and shikimate dehydrogenase are encoded by the E. coli aroF FBR , E. coli aroB, and E. coli aroE genes, respectively; wherein

(a) at least one of said enzymes are encoded by recombinant DNA,

(b) the recombinant microbe comprises an inactivating mutation in an endogenous DNA encoding a dehydroquinate dehydratase, and

(c) the recombinant microbe is capable of producing quinic acid from a carbon source by using all three enzymes,

wherein the recombinant microbe is cultured to allow all three enzymes to catalyze the reactions to synthesize quinic acid.

6. The method according to claim 2 , wherein the recombinant microbe is a recombinant E. coli.

7. The method according to claim 1 , wherein the E. coli aroE gene is the same aroE gene found in plasmid pKD12.112.

8. The method according to claim 2 , wherein the inactivating mutation is in an E. coli aroD gene.

9. The method according to claim 8 , wherein the recombinant microbe is E. coli QP1.1.