Compositions and methods for the biosynthesis of vanillin or vanillin beta-d-glucoside

Abstract

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express a mutant AROM polypeptide and/or mutant catechol-O-methyltransferase polypeptide alone or in combination with one or more vanillin biosynthetic enzymes or UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce vanillin or vanillin beta-D-glucoside.

What is claimed is: 
     
         1 . A method for producing vanillin and/or vanillin beta-D-glucoside comprising
 (a) providing a recombinant host capable of producing vanillin, wherein said recombinant host harbors a heterologous nucleic acid encoding  Rhodococcus jostii, Rhodococcus opacus  or Modestobacter  marinus  vanillyl alcohol oxidase; and   (b) cultivating said recombinant host for a time sufficient for said recombinant host to produce vanillin and/or vanillin glucoside, wherein said recombinant host exhibits a reduction in the accumulation of vanillyl alcohol.   
     
     
         2 . The method of  claim 1 , wherein said host is a microorganism. 
     
     
         3 . The method of  claim 2 , wherein said host is  Saccharomyces cerevisiae, Schizosaccharomyces pombe  or  Escherichia coli.    
     
     
         4 . The method of  claim 1 , wherein said host is a plant or plant cell. 
     
     
         5 . The method of  claim 1 , wherein said host further comprises a gene encoding a 3-dehydroshikimate dehydratase (3DSD), a gene encoding a aromatic carboxylic acid reductase (ACAR), a gene encoding a uridine 5′-diphosphoglucosyl transferase (UGT), a gene encoding a phosphopantetheine transferase (PPTase) and/or a gene encoding a catechol-O-methyl transferase (COMT). 
     
     
         6 . An extract comprising vanillin and/or vanillin glucoside produced by the method of  claim 1 . 
     
     
         7 . A consumable comprising the extract of  claim 6 . 
     
     
         8 . The consumable of  claim 7 , wherein the consumable is a food product, pharmaceutical composition, a dietary supplement, a nutraceutical, a dental hygienic composition, a tabletop sweetener, or a cosmetic product. 
     
     
         9 . A recombinant host comprising a heterologous nucleic acid encoding  Rhodococcus jostii, Rhodococcus opacus  or  Modestobacter marinas  vanillyl alcohol oxidase. 
     
     
         10 . The recombinant host of  claim 9 , wherein the recombinant host further comprises a nucleic acid encoding a 3-dehydroshikimate dehydratase (3DSD) polypeptide, a nucleic acid encoding an aromatic carboxylic acid reductase (ACAR) polypeptide, a nucleic acid encoding a phosphopantetheine transferase (PPTase) polypeptide, a nucleic acid encoding an uridine 5′-diphosphoglucosyl transferase (UGT) polypeptide and/or a nucleic acid encoding catechol-O-methyl transferase (COMT). 
     
     
         11 . The recombinant host of  claim 9 , wherein the recombinant host is a microorganism. 
     
     
         12 . The recombinant host of  claim 11 , wherein the recombinant host is  Saccharomyces cerevisiae, Schizosaccharomyces pombe  or  Escherichia coli.    
     
     
         13 . The recombinant host of  claim 9 , wherein said host is a plant or plant cell.