Terephthalic acid producing proteobacteria

Abstract

This invention relates to a biocatalytic process to produce terephthalic acid and isophthalic acid from p-xylene and m-xylene, respectively. Terephthalic acid has been prepared by oxidizing p-xylene with bacteria belonging to the genus Burkholderia. Conversion of p-xylene into terephthalic acid is accomplished by a single bacterial strain that produces all of the requisite enzymes. In addition, this invention relates to the preparation of isophthalic acid from a mixture of m- and p-xylene.

What is claimed is:  
     
         1 . A TPA producing microorganism isolated by the process comprising: 
 (i) culturing a sample suspected of containing a TPA producing microorganism in a suitable growth medium containing at least one aromatic organic substrate selected from the group consisting of p-xylene, p-toluic acid, and terephthalic acid;    (ii) selecting those microorganisms which are able to use every substrate selected individually from the group consisting of p-xylene, p-toluic acid, and terephthalic acid as a sole carbon source;    (iii) contacting the microorganisms selected in step (ii) with p-xylene to form a reaction medium; and    (iv) monitoring the reaction medium over time for the presence of terephthalic acid, whereby the production of terephthalic acid indicates the presence of a TPA producing microorganism.    
     
     
         2 . The TPA producing microorganism of  claim 1  wherein the microorganism comprises the genes encoding the TPA biosynthetic pathway  
     
     
         3 . The TPA producing microorganism of  claim 2  wherein the microorganism is a Proteobacteria.  
     
     
         4 . The TPA producing microorganism of  claim 3  selected from the group consisting of, Burkholderia, Alcaligenes, Pseudomonas, Shphingomonas and Comamonas.  
     
     
         5 . The TPA producing microorganism of  claim 4  selected from the group consisting of IR3 (ATCC 202150) and IR10 (ATCC 202151).  
     
     
         6 . The TPA producing microorganism of claims  3  or  4  wherein the 16s RNA of the TPA producing microorganism has at least 95% identity with the 16s rRNA sequence set forth in SEQ ID NO:1.  
     
     
         7 . A process for the isolation of a TPA producing microorganism comprising: 
 (i) culturing a sample suspected of containing a TPA producing microorganism in a suitable growth medium containing at least one aromatic organic substrate selected from the group consisting of p-xylene, p-toluic acid, and terephthalic acid;    (ii) selecting those microorganisms which are able to use every substrate selected individually from the group consisting of p-xylene, p-toluic acid, and terephthalic acid as a sole carbon source;    (iii) contacting the microorganisms selected in step (ii) with p-xylene to form a reaction medium; and    (iv) monitoring the reaction medium over time for the presence of terephthalic acid, whereby the production of terephthalic acid indicates the presence of a TPA producing microorganism.    
     
     
         8 . A process for the production of terephthalic acid comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate whereby terephthalic acid accumulates; and    (ii) optionally recovering the terephthalic acid.    
     
     
         9 . The process of  claim 8  wherein the aromatic organic substrate is selected from the group consisting of p-xylene, 4-methylbenzyl alcohol, p-tolualdehyde, p-toluic acid, 4-carboxybenzyl alcohol, and 4-carboxybenzaldehyde.  
     
     
         10 . The process of  claim 8  wherein the TPA producing microorganism is isolated by the process of  claim 7 .  
     
     
         11 . The process of  claim 10  wherein the isolated TPA producing microorganism is a Proteobacteria.  
     
     
         12 . The process of  claim 11  wherein the isolated TPA producing microorganism is selected from the group consisting of, Burkholderia, Alcaligenes, Pseudomonas, Shphingomonas and Comamonas.  
     
     
         13 . The process of  claim 12  wherein the 16s RNA of the TPA producing microorganism has at least 97% identity with the 16s rRNA sequence set forth in SEQ ID NO:1.  
     
     
         14 . The process of  claim 13  wherein the isolated TPA producing microorganism is Burkholderia sp. selected from the group consisting of IR3 (ATCC 202150) and IR10 (ATCC 202151).  
     
     
         15 . A process for the production of 4-carboxybenzyl alcohol comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate whereby 4-carboxybenzyl alcohol accumulates; and    (ii) optionally recovering the 4-carboxybenzyl alcohol.    
     
     
         16 . A process for the production of 4-carboxybenzaldehyde comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate whereby 4-carboxybenzaldehyde accumulates; and    (ii) optionally recovering the 4-carboxybenzaldehyde.    
     
     
         17 . A process for the production of 1,4-benzenedimethanol comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate whereby 1,4-benzenedimethanol accumulates; and    (ii) optionally recovering the 1,4-benzenedimethanol.    
     
     
         18 . A process for the production of terephthalaldehyde comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate whereby terephthalaldehyde accumulates; and    (ii) optionally recovering the terephthalaldehyde.    
     
     
         19 . The process of either of claims  15 ,  16 ,  17  or  18  wherein the isolated TPA producing microorganism is isolated by the process of  claim 7 .  
     
     
         20 . The process of  claim 19  wherein the isolated TPA producing microorganism is Burkholderia sp. selected from the group consisting of IR3 (ATCC 202150) and IR10 (ATCC 202151).  
     
     
         21 . The process of either of claims  15 ,  16 ,  17  or  18  wherein the aromatic organic substrate is selected from the group consisting of p-xylene, 4-methylbenzyl alcohol, p-tolualdehyde, p-toluic acid, and 4-carboxybenzyl alcohol.  
     
     
         22 . A process for the production of isophthalic acid comprising: 
 (i) contacting an isolated TPA producing microorganism with an aromatic organic substrate and at least one suitable induction compound whereby isophthalic acid accumulates; and    (ii) optionally recovering the isophthalic acid.    
     
     
         23 . The process of  claim 22  wherein the isolated TPA producing microorganism is isolated by the process of  claim 7 .  
     
     
         24 . The process of  claim 23  wherein the isolated TPA producing microorganism is a Proteobacteria.  
     
     
         25 . The process of  claim 24  wherein the isolated TPA producing microorganism is selected from the group consisting of, Burkholderia, Alcaligenes, Pseudomonas, Shphingomonas and Comamonas.  
     
     
         26 . The process of  claim 25  wherein the 16s RNA of the TPA producing microorganism has at least 97% identity with the 16s rRNA sequence set forth in SEQ ID NO:1.  
     
     
         27 . The process of  claim 26  wherein the isolated TPA producing microorganism is Burkholderia sp. selected from the group consisting of IR3 (ATCC 202150) and IR10 (ATCC 202151).  
     
     
         28 . The process of  claim 22  wherein the aromatic organic substrate is selected from the group consisting of m-xylene, 3-methylbenzyl alcohol, m-tolualdehyde, m-toluic acid, 3-carboxybenzyl alcohol, and 3-carboxybenzaldehyde.  
     
     
         29 . The process of  claim 22  wherein the suitable induction compound is selected from the group consisting of p-xylene, 4-methylbenzyl alcohol, p-tolualdehyde, p-toluic acid, 4-carboxybenzyl alcohol, 4-carboxybenzaldehyde and terephthalic acid.  
     
     
         30 . A process for the production of terephthalic acid comprising: 
 (i) contacting a mixed population of microorganisms comprising the genes encoding the TPA biosynthetic pathway, with an aromatic organic substrate whereby terephthalic acid accumulates; and    (ii) optionally recovering the terephthalic acid.    
     
     
         31 . The process of  claim 28  wherein the aromatic organic substrate is selected from the group consisting of p-xylene, 4-methylbenzyl alcohol, p-tolualdehyde, p-toluic acid, 4-carboxybenzyl alcohol, and 4-carboxybenzaldehyde.