Methods for producing isobutene from 3-methylcrotonic acid

Abstract

Described are methods for the production of isobutene comprising the enzymatic conversion of 3-methylcrotonic acid into isobutene wherein said 3-methylcrotonic acid is obtained by the enzymatic conversion of 3-methylcrotonyl-CoA into 3-methylcrotonic acid or wherein said 3-methylcrotonic acid is obtained by the enzymatic conversion of 3-hydroxyisovalerate (HIV) into 3-methylcrotonic acid. It is described that the enzymatic conversion of 3-methylcrotonic acid into isobutene can, e.g., be achieved by making use of a 3-methylcrotonic acid decarboxylase, preferably an FMN-dependent decarboxylase associated with an FMN prenyl transferase, an aconitate decarboxylase (EC 4.1.1.6), a methylcrotonyl-CoA carboxylase (EC 6.4.1.4), or a geranoyl-CoA carboxylase (EC 6.4.1.5).

1 - 36 . (canceled) 
     
     
         37 . A recombinant organism or microorganism capable of producing isobutene, wherein said microorganism expresses polypeptides comprising:
 a) at least one of:
 i. a CoA transferase (EC 2.8.3.-) and a thioester hydrolase (EC 3.1.2.-); or 
 ii. (a) a phosphate butyryltransferase (EC 2.3.1.19) or a phosphate acetyltransferase (EC 2.3.1.8) and
 (b) a phosphotransferase with a carboxy group as acceptor (EC 2.7.2.-) 
 
   and   b) a 3-methylcrotonic acid decarboxylase selected from:
 i. an FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-); 
 ii. an aconitate decarboxylase (EC 4.1.1.6); 
 iii. a methylcrotonyl-CoA carboxylase (EC 6.4.1.4); 
 iv. a geranoyl-CoA carboxylase (EC 6.4.1.5); or 
 v. a protocatechuate (PCA) decarboxylase (EC 4.1.1.63). 
   
     
     
         38 . The recombinant organism or microorganism of  claim 37 , wherein the CoA transferase (EC 2.8.3.-) is selected from a propionate:acetate-CoA transferase (EC 2.8.3.1), an acetate CoA-transferase (EC 2.8.3.8) or a succinyl-CoA:acetate CoA-transferase (EC 2.8.3.18). 
     
     
         39 . The recombinant organism or microorganism of  claim 37 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         40 . The recombinant organism or microorganism of  claim 37 , wherein the phosphotransferase with a carboxy group as acceptor (EC 2.7.2.-) is selected from a propionate kinase (EC 2.7.2.15), an acetate kinase (EC 2.7.2.1), a butyrate kinase (EC 2.7.2.7) or a branched-chain-fatty-acid kinase (EC 2.7.2.14). 
     
     
         41 . The recombinant organism or microorganism of  claim 37 , wherein the recombinant organism or microorganism further expresses a polypeptide selected from a methylcrotonyl-CoA carboxylase (EC 6.4.1.4) or a geranoyl-CoA carboxylase (EC 6.4.1.5). 
     
     
         42 . The recombinant organism or microorganism of  claim 41 , wherein the recombinant organism or microorganism further expresses a polypeptide selected from a 3-methylglutaconyl-coenzyme A hydratase (EC 4.2.1.18), a 3-hydroxyacyl-CoA dehydratase (EC 4.2.1.-) or an enoyl-CoA hydratase (EC 4.2.1.-). 
     
     
         43 . The recombinant organism or microorganism of  claim 42 , wherein the recombinant organism or microorganism further expresses a polypeptide selected from a 3-hydroxy-3-methylglutaryl-CoA synthase. 
     
     
         44 . The recombinant organism or microorganism of  claim 37 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         45 . The recombinant organism or microorganism of  claim 38 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         46 . The recombinant organism or microorganism of  claim 39 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         47 . The recombinant organism or microorganism of  claim 46 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         48 . The recombinant organism or microorganism of  claim 40 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         49 . The recombinant organism or microorganism of  claim 48 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         50 . The recombinant organism or microorganism of  claim 41 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         51 . The recombinant organism or microorganism of  claim 50 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         52 . The recombinant organism or microorganism of  claim 42 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         53 . The recombinant organism or microorganism of  claim 52 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         54 . The recombinant organism or microorganism of  claim 43 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         55 . The recombinant organism or microorganism of  claim 54 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         56 . The recombinant organism or microorganism of  claim 44 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         57 . The recombinant organism or microorganism of  claim 56 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20). 
     
     
         58 . The recombinant organism or microorganism of  claim 45 , wherein the 3-methylcrotonic acid decarboxylase is a FMN-dependent decarboxylase associated with an FMN prenyl transferase (EC 2.5.1.-). 
     
     
         59 . The recombinant organism or microorganism of  claim 58 , wherein the thioester hydrolase (EC 3.1.2.-) is selected from an acetyl-CoA hydrolase (EC 3.1.2.1), an ADP-dependent short-chain-acyl-CoA hydrolase (EC 3.1.2.18) or an acyl-CoA hydrolase (EC 3.1.2.20).