Microorganisms and methods for the biosynthesis of butadiene

Abstract

The invention provides non-naturally occurring microbial organisms having a butadiene pathway. The invention additionally provides methods of using such organisms to produce butadiene.

1 . A non-naturally occurring microbial organism, comprising a microbial organism having a butadiene pathway comprising at least one exogenous nucleic acid encoding a butadiene pathway enzyme expressed in a sufficient amount to produce butadiene, said butadiene pathway comprising a butadiene synthase, an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a crotonyl-CoA hydrolase, synthetase, or transferase, a crotonate reductase, a crotonyl-CoA reductase (alcohol forming), a glutaconyl-CoA decarboxylase, a glutaryl-CoA dehydrogenase, an 3-aminobutyryl-CoA deaminase, a 4-hydroxybutyryl-CoA dehydratase or a crotyl alcohol diphosphokinase. 
     
     
         2 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises two exogenous nucleic acids each encoding a butadiene pathway enzyme. 
     
     
         3 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises three exogenous nucleic acids each encoding a butadiene pathway enzyme. 
     
     
         4 . The non-naturally occurring microbial organism of  claim 1 , wherein said microbial organism comprises four exogenous nucleic acids each encoding a butadiene pathway enzyme. 
     
     
         5 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase and a butadiene synthase. 
     
     
         6 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase and crotonyl-CoA reductase (alcohol forming). 
     
     
         7 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a butadiene synthase, a crotonyl-CoA reductase (alcohol forming) and a crotyl alcohol diphosphokinase. 
     
     
         8 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase, a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase. 
     
     
         9 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a butadiene synthase, a crotonyl-CoA hydrolase, synthetase or transferase, a crotonate reductase and a crotyl alcohol diphosphokinase. 
     
     
         10 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a butadiene synthase and a crotyl alcohol diphosphokinase. 
     
     
         11 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaconyl-CoA decarboxylase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase and a butadiene synthase. 
     
     
         12 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaconyl-CoA decarboxylase, a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase and crotonyl-CoA reductase (alcohol forming). 
     
     
         13 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaconyl-CoA decarboxylase, a butadiene synthase, a crotonyl-CoA reductase (alcohol forming) and a crotyl alcohol diphosphokinase. 
     
     
         14 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaconyl-CoA decarboxylase, a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase, a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase. 
     
     
         15 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaconyl-CoA decarboxylase, a crotonaldehyde reductase (alcohol forming), a butadiene synthase, a crotonyl-CoA hydrolase, synthetase or transferase, a crotonate reductase and a crotyl alcohol diphosphokinase. 
     
     
         16 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a butadiene a glutaconyl-CoA decarboxylase and a crotyl alcohol diphosphokinase. 
     
     
         17 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaryl-CoA dehydrogenase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase and a butadiene synthase. 
     
     
         18 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaryl-CoA dehydrogenase, a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase and crotonyl-CoA reductase (alcohol forming). 
     
     
         19 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaryl-CoA dehydrogenase, a butadiene synthase, a crotonyl-CoA reductase (alcohol forming) and a crotyl alcohol diphosphokinase. 
     
     
         20 . The non-naturally occurring microbial organism of  claim 1 , wherein said butadiene pathway comprises a glutaryl-CoA dehydrogenase, a crotonaldehyde reductase (alcohol forming), a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase, a butadiene synthase, a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase. 
     
     
         21 - 59 . (canceled)