Production of p-hydroxybenzoic acid

Abstract

One aspect of the present invention relates to a transgenic plant which, by presence of a transgene, is able to produce p-hydroxybenzoic acid or a β-D-glycoside or β-D-glucose ester thereof. A method is also disclosed for producing p-hydroxybenzoic acid or a β-D-glycoside or β-D-glucose ester thereof using a transgenic plant of the present invention.

67 . A transgenic plant which, by presence of a transgene, is able to produce p-hydroxybenzoic acid or a β-D-glycoside or β-D-glucose ester thereof.  
     
     
         68 . A transgenic plant according to  claim 67  which is transgenic for a polynucleotide encoding trans-ferulate:CoASH ligase activity, trans-feruloyl ScoA activity, 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity, or vanillin:NAD + oxidoreductase activity.  
     
     
         69 . A transgenic plant according to  claim 68  wherein the vanillin:NAD + oxidoreductase activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 4.  
     
     
         70 . A transgenic plant according to  claim 68  wherein the 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 2.  
     
     
         71 . A transgenic plant according to  claim 68  wherein the trans-feruloyl ScoA activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 2.  
     
     
         72 . A transgenic plant according to  claim 68  wherein the plant produces p-hydroxybenzoic acid.  
     
     
         73 . A transgenic plant according to  claim 68  wherein the plant produces a β-D-glycoside or a β-D-glucose ester of p-hydroxybenzoic acid.  
     
     
         74 . A transgenic plant according to  claim 68  wherein the polynucleotide encodes 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity in a location in the plant which contains 4-trans-coumaroyl SCoA.  
     
     
         75 . A transgenic plant according to  claim 68  wherein said plant is selected from Nicotiana spp.,  Solanum tuberosum,  Brassica spp., Beta spp., Capsicum spp. and Vanilla spp.  
     
     
         76 . A method of producing p-hydroxybenzoic acid or a β-D-glycoside or β-D-glucose ester thereof, the method comprising: 
 providing a transgenic plant according to  claim 67  under conditions effective to convert 4-trans-coumaric acid, salts thereof, or 4-trans-coumaroyl SCoA to p-hydroxybenzoic acid or a β-D-glycoside or β-D-glucose ester thereof.  
 
     
     
         77 . A method according to  claim 76  wherein the transgenic plant comprises a heterologous polynucleotide encoding trans-ferulate:CoASH ligase activity, trans-feruloyl ScoA activity, 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity, or vanillin:NAD+oxidoreductase activity.  
     
     
         78 . A method according to  claim 77  wherein the vanillin:NAD +  oxidoreductase activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 4.  
     
     
         79 . A method according to  claim 77  wherein the 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 2.  
     
     
         80 . A method according to  claim 77  wherein the trans-feruloyl ScoA activity is afforded by a protein or polypeptide having the amino acid sequence of SEQ ID No: 2.  
     
     
         81 . A method according to  claim 76  wherein p-hydroxybenzoic acid is produced.  
     
     
         82 . A method according to  claim 76  wherein a β-D-glycoside or β-D-glucose ester of p-hydroxybenzoic acid is produced.  
     
     
         83 . A method according to  claim 82  further comprising: 
 treating the β-D-glycoside or β-D-glucose ester of p-hydroxybenzoic acid via acid or base-catalysed hydrolysis under conditions effective form p-hydroxybenzoic acid.  
 
     
     
         84 . A method according to  claim 82  further comprising: 
 treating the β-D-glycoside or β-D-glucose ester of p-hydroxybenzoic acid with a β-D-glucosidase under conditions effective to form p-hydroxybenzoic acid.  
 
     
     
         85 . A method according to  claim 76  wherein the polynucleotide encodes 4-hydroxy-3-methoxyphenyl-β-hydroxy-propionyl ScoA activity in a location in the plant which contains 4-trans-coumaroyl SCoA.  
     
     
         86 . A method according to  claim 76  wherein the transgenic plant is selected from Nicotiana spp.,  Solanum tuberosum,  Brassica spp., Beta spp., Capsicum spp. and Vanilla spp.