Microorganisms for producing 4c-5c compounds with unsaturation and methods related thereto

Abstract

The invention provides a non-naturally occurring microbial organism having a butadiene, crotyl alcohol, 2,4-pentadienoate, 3-buten-2-ol, or 3-buten-1-ol, pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in a pathway. The invention additionally provides a method for producing butadiene, crotyl alcohol, 2,4-pentadienoate, 3-buten-2-ol, or 3-buten-1-ol. The method can include culturing a butadiene, crotyl alcohol, 2,4-pentadienoate, 3-buten-2-ol, or 3-buten-1-ol-producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding a pathway enzyme in a sufficient amount, and under conditions and for a sufficient period of time to produce butadiene, crotyl alcohol, 2,4-pentadienoate, 3-buten-2-ol, or 3-buten-1-ol.

1 - 23 . (canceled) 
     
     
         24 . A non-naturally occurring microbial organism having a pathway to butadiene, crotyl alcohol, 2,4-pentadienoate, or 3-buten-1-ol, said microbial organism comprising at least one exogenous nucleic acid encoding a pathway enzyme, the enzyme selected from the group consisting of (2E) 2-hydroxypent-4-enoate vinylisomerase, (2H) 2-hydroxypent-4-enoate mutase, and (2M) 3-hydroxypent-4-enoyl-CoA vinylisomerase. 
     
     
         25 . The non-naturally occurring microorganism of  claim 24 , further comprising a nucleic acid encoding an enzyme selected from an alcohol dehydrogenase, an acyl-CoA reductase, a CoA transferase, a CoA hydrolase, a decarboxylase, an aldolase, a dehydratase, an isomerase, a CoA synthetase, an alcohol mutase, or combinations thereof. 
     
     
         26 . The non-naturally occurring microorganism of  claim 24 , further comprising one or more nucleic acid(s) encoding an enzyme selected from (2A) acetyl-CoA reductase, (2B) 4-hydroxy 2-oxovalerate aldolase, (2C) 4-hydroxy 2-oxovalerate dehydratase, and (2D) 2-oxopent-4-enoate reductase. 
     
     
         27 . The non-naturally occurring microorganism of  claim 24 , wherein the enzyme is (2E) 2-hydroxypent-4-enoate vinylisomerase. 
     
     
         28 . The non-naturally occurring microorganism of  claim 27 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2P) 5-hydroxypent-2-enoate decarboxylase and (2Q) 3-buten-1-ol dehydratase or further comprising a nucleic acid encoding one or more enzyme(s) selected from (2P) 5-hydroxypent-2-enoate decarboxylase and (2S) vinylisomerase. 
     
     
         29 . The non-naturally occurring microorganism of  claim 27 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2F) 5-hydroxypent-2-enoate dehydratase and (2G) 2,4-pentadienoate decarboxylase. 
     
     
         30 . The non-naturally occurring microorganism of  claim 24 , wherein the enzyme is (2H) 2-hydroxypent-4-enoate mutase. 
     
     
         31 . The non-naturally occurring microorganism of  claim 30 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2R) 3-hydroxypent-4-enoate dehydratase and (2G) 2,4-pentadienoate decarboxylase. 
     
     
         32 . The non-naturally occurring microorganism of  claim 30 , further comprising a nucleic acid encoding (2K) 3-hydroxypent-4-enoate decarboxylase. 
     
     
         33 . The non-naturally occurring microorganism of  claim 30 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2L) 3-hydroxypent-4-enoyl-CoA synthetase or transferase, (2M) 3-hydroxypent-4-enoyl-CoA vinylisomerase, (2 I 5-hydroxypent-2-enoyl-CoA synthetase, transferase or hydrolase (2F) 5-hydroxypent-2-enoate dehydratase, and (2G) 2,4-pentadienoate decarboxylase. 
     
     
         34 . The non-naturally occurring microorganism of  claim 30 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2L) 3-hydroxypent-4-enoyl-CoA synthetase or transferase, (2M) 3-hydroxypent-4-enoyl-CoA vinylisomerase, (2N) 2,4-pentadienoyl-CoA synthetase, (2 J) 2,4-pentadienoyl-CoA synthetase, transferase or hydrolase, and (2G) 2,4-pentadienoate decarboxylase. 
     
     
         35 . The non-naturally occurring microorganism of  claim 30 , further comprising a nucleic acid encoding one or more enzyme(s) selected from (2L) 3-hydroxypent-4-enoyl-CoA synthetase or transferase, (2O) 3-hydroxypent-4-enoyl-CoA dehydratase, (2 J) 2,4-pentadienoyl-CoA synthetase, transferase or hydrolase, and (2G) 2,4-pentadienoate decarboxylase. 
     
     
         36 . The non-naturally occurring microbial organism of  claim 24  having a pathway to convert pyruvate and acetyl-CoA to butadiene, crotyl alcohol, 2,4-pentadienoate, or 3-buten-1-ol. 
     
     
         37 . The non-naturally occurring microorganism of  claim 24  wherein any one or more nucleic acid(s) is an exogenous nucleic acid. 
     
     
         38 . A method for the production of butadiene, crotyl alcohol, 2,4-pentadienoate, or 3-buten-1-ol, comprising culturing the non-naturally occurring microorganism of  claim 24  under conditions to produce butadiene, crotyl alcohol, 2,4-pentadienoate, or 3-buten-1-ol. 
     
     
         39 . The method for the production of butadiene, crotyl alcohol, 2,4-pentadienoate, or 3-buten-1-ol of  claim 38 , wherein the culturing comprises substantially anaerobic conditions. 
     
     
         40 . A non-naturally occurring microbial organism having a pathway to butadiene, 2,4-pentadienoate, or 3-buten-2-ol, said microbial organism comprising at least one exogenous nucleic acid encoding a pathway enzyme, the enzyme selected from the group consisting of (3D) 3,4-dihydroxypentanoate dehydratase, (3E) 4-hydroxypent-2-enoate decarboxylase (3H) 3,4-dihydroxypentanoyl-CoA dehydratase, and (3J) 4-hydroxypent-2-enoyl-CoA transferase. 
     
     
         41 - 44 . (canceled) 
     
     
         45 . A non-naturally occurring microbial organism having a pathway to butadiene or 3-buten-2-ol, said microbial organism comprising at least one exogenous nucleic acid encoding a pathway enzyme, the enzyme selected from the group consisting of (4D) 3-buten-2-ol synthase, (4E) 3-buten-2-ol synthase, and (4F) crotyl alcohol isomerase. 
     
     
         46 - 72 . (canceled)