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 . A non-naturally occurring microbial organism, comprising a microbial organism having an aniline pathway comprising at least one exogenous nucleic acid encoding an aniline pathway enzyme expressed in a sufficient amount to produce aniline, said aniline pathway comprising a 4-aminobenzoate carboxylyase, an aminodeoxychorismate synthase, and an aminodeoxychorismate lyase. 
     
     
         2 . The non-naturally occurring microbial organism of  claim 1 , further comprising a DAHP synthase. 
     
     
         3 . The non-naturally occurring microbial organism of  claim 1 , further comprising an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         4 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises two exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         5 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         6 . The non-naturally occurring microbial organism of  claim 5 , wherein said three exogenous nucleic acids encode an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, and a 4-aminobenzoate carboxylyase. 
     
     
         7 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         8 . The non-naturally occurring microbial organism of  claim 7 , wherein said four exogenous nucleic acids encode a DAHP synthase, an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, and a 4-aminobenzoate carboxylyase. 
     
     
         9 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises five exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         10 . The non-naturally occurring microbial organism of  claim 9 , wherein said five exogenous nucleic acids encode, a DAHP synthase, an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, and a 4-aminobenzoate carboxylyase and an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         11 . The non-naturally occurring microbial organism of  claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid. 
     
     
         12 . The non-naturally occurring microbial organism of  claim 1 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 
     
     
         13 . A method for producing aniline, comprising culturing the non-naturally occurring microbial organism of  claim 1  under conditions and for a sufficient period of time to produce aniline. 
     
     
         14 . The method of  claim 13  further comprising a DAHP synthase. 
     
     
         15 . The method of  claim 13 , further comprising an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         16 . The method of  claim 13 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 
     
     
         17 . The method of  claim 13 , wherein said microbial organism comprises two exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         18 . The method of  claim 13 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         19 . The method of  claim 18 , wherein said three exogenous nucleic acids encode a 4-aminobenzoate carboxylyase, an aminodeoxychorismate synthase, and an aminodeoxychorismate lyase. 
     
     
         20 . The method of  claim 13 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         21 . The method of  claim 20 , wherein said four exogenous nucleic acids encode a DAHP synthase, an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, and a 4-aminobenzoate carboxylyase. 
     
     
         22 . The method of  claim 13 , wherein said microbial organism comprises five exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         23 . The method of  claim 22 , wherein said five exogenous nucleic acids encode a DAHP synthase, an aminodeoxychorismate synthase, an aminodeoxychorismate lyase, a 4-aminobenzoate carboxylyase and an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         24 . The method of  claim 13 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid. 
     
     
         25 . A non-naturally occurring microbial organism, comprising a microbial organism having an aniline pathway comprising at least one exogenous nucleic acid encoding an aniline pathway enzyme expressed in a sufficient amount to produce aniline, said aniline pathway comprising an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         26 . The non-naturally occurring microbial organism of  claim 25 , further comprising a DAHP synthase. 
     
     
         27 . The non-naturally occurring microbial organism of  claim 25 , further comprising an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         28 . The non-naturally occurring microbial organism of  claim 25 , wherein said microbial organism comprises two exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         29 . The non-naturally occurring microbial organism of  claim 28 , wherein said two exogenous nucleic acids encode an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         30 . The non-naturally occurring microbial organism of  claim 25 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         31 . The non-naturally occurring microbial organism of  claim 30 , wherein said three exogenous nucleic acids encode a DAHP synthase, an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         32 . The non-naturally occurring microbial organism of  claim 25 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         33 . The non-naturally occurring microbial organism of  claim 32 , wherein said four exogenous nucleic acids encode a 3-dehydroquinate synthase, a DAHP synthase, an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         34 . The non-naturally occurring microbial organism of  claim 25 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid. 
     
     
         35 . The non-naturally occurring microbial organism of  claim 25 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 
     
     
         36 . A method for producing aniline, comprising culturing the non-naturally occurring microbial organism of  claim 25  under conditions and for a sufficient period of time to produce aniline. 
     
     
         37 . The method of  claim 36  further comprising a DAHP synthase. 
     
     
         38 . The method of  claim 36 , further comprising an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         39 . The method of  claim 36 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium. 
     
     
         40 . The method of  claim 36 , wherein said microbial organism comprises two exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         41 . The method of  claim 40 , wherein said two exogenous nucleic acids encode an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         42 . The method of  claim 36 , wherein said microbial organism comprises three exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         43 . The method of  claim 42 , wherein said three exogenous nucleic acids encode a DAHP synthase, an anthranilate synthase and an anthranilate decarboxylase. 
     
     
         44 . The method of  claim 36 , wherein said microbial organism comprises four exogenous nucleic acids each encoding an aniline pathway enzyme. 
     
     
         45 . The method of  claim 44 , wherein said four exogenous nucleic acids encode a DAHP synthase, an anthranilate synthase, an anthranilate decarboxylase and an enzyme selected from a 3-dehydroquinate synthase, a 3-dehydroquinate dehydratase, a shikimate dehydrogenase or a quinate/shikimate dehydrogenase, a shikimate kinase, a 3-phosphoshikimate-1-carboxyvinyltransferase, and a chorismate synthase. 
     
     
         46 . The method of  claim 36 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.