Microorganisms for the production of aniline

Abstract

A non-naturally occurring microbial organism having an aniline pathway includes at least one exogenous nucleic acid encoding an aniline pathway enzyme expressed in a sufficient amount to produce aniline. The aniline pathway includes (1) an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, and a 4-aminobenzoate carboxylyase or (2) an anthranilate synthase and an anthranilate decarboxylase. A method for producing aniline, includes culturing these non-naturally occurring microbial organisms under conditions and for a sufficient period of time to produce aniline.

1 - 46 . (canceled) 
     
     
         47 . A non-naturally occurring  Escherichia coli , comprising an aniline pathway, said aniline pathway comprising an anthranilate synthase, a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, a chorismate synthase and an anthranilate decarboxylase, wherein said non-naturally occurring  Escherichia coli , comprises at least two exogenous nucleic acids, the two exogenous nucleic acids encoding enzymes selected from the group consisting of the 3-dehydroquinate synthase, the anthranilate synthase, the 3-dehydroquinate dehydratase, the shikimate dehydrogenase or the quinate/shikimate dehydrogenase, the shikimate kinase, the 3-phosphoshikimate-1-carboxyvinyltransferase, and the chorismate synthase, wherein an endogenous nucleic acid encodes the anthranilate decarboxylase, and wherein the aniline pathway enzymes are expressed in a sufficient amount to produce aniline. 
     
     
         48 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein the aniline pathway further comprises a 3-deoxy-D-arabino-heptulosonic acid-7-phosphate (DAHP) synthase. 
     
     
         49 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         50 . The non-naturally occurring  Escherichia coli  of  claim 49 , wherein said three exogenous nucleic acids encode the DAHP synthase, the anthranilate synthase and the 3-dehydroquinate synthase. 
     
     
         51 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         52 . The non-naturally occurring  Escherichia coli  of  claim 51  wherein said four exogenous nucleic acids encode the 3-dehydroquinate synthase, the DAHP synthase, the anthranilate synthase and the chorismate synthase. 
     
     
         53 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid. 
     
     
         54 . An anaerobic culture medium comprising the microbial organism of  claim 47 . 
     
     
         55 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises five exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         56 . The non-naturally occurring  Escherichia coli  of  claim 55 , wherein said five exogenous nucleic acids encoding the anthranilate synthase, the DAHP synthase, the 3-dehydroquinate synthase, the 3-dehydroquinate dehydratase, the shikimate dehydrogenase or the quinate/shikimate dehydrogenase. 
     
     
         57 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises six exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         58 . The non-naturally occurring  Escherichia coli  of  claim 57 , wherein said six exogenous nucleic acids encoding the anthranilate synthase, the DAHP synthase, the 3-dehydroquinate synthase, the 3-dehydroquinate dehydratase, the shikimate dehydrogenase or the quinate/shikimate dehydrogenase and a shikimate kinase. 
     
     
         59 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises seven exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         60 . The non-naturally occurring  Escherichia coli  of  claim 59 , wherein said seven exogenous nucleic acids encoding the anthranilate synthase, the DAHP synthase, the 3-dehydroquinate synthase, the 3-dehydroquinate dehydratase, the shikimate dehydrogenase or the quinate/shikimate dehydrogenase, the shikimate kinase, the 3-phosphoshikimate-1carboxyvinyltransferase. 
     
     
         61 . The non-naturally occurring  Escherichia coli  of  claim 47 , wherein said microbial organism comprises eight exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         62 . The non-naturally occurring  Escherichia coli  of  claim 61 , wherein said eight exogenous nucleic acids encoding the anthranilate synthase, the DAHP synthase, the 3-dehydroquinate synthase, the 3-dehydroquinate dehydratase, the shikimate dehydrogenase or the quinate/shikimate dehydrogenase the shikimate kinase, the 3-phosphoshikimate-1-carboxyvinyltransferase, and the chorismate synthase.