Adipate (ester or thioester) synthesis

Abstract

The present invention relates to a method for preparing an adipate ester or thioester. The invention further relates to a method for preparing adipic acid from said ester or thioester. Further the invention provides a number of methods for preparing an intermediate for said ester or thioester. Further the invention relates to a method for preparing 6-amino caproic acid (6-ACA), a method for preparing 5-formyl valeric acid (5-FVA), and a method for preparing caprolactam. Further, the invention relates to a host cell for use in a method according to the invention.

1 - 19 . (canceled) 
     
     
         20 . A method for preparing 6-amino caproic acid, comprising converting the 5-formylpentanoate into 6-amino caproic acid in the presence of a biocatalyst, wherein the biocatalyst comprises an enzyme, wherein said enzyme comprises the amino acid sequence of any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138, or an amino acid sequence that has at least 90% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136 and 138, and wherein said enzyme catalyzes the conversion of 5-formylpentanoate into 6-amino caproic acid. 
     
     
         21 - 31 . (canceled) 
     
     
         32 . The method of  claim 20 , wherein said enzyme comprises an amino acid sequence that has at least 92% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         33 . The method of  claim 20 , wherein said enzyme comprises an amino acid sequence that has at least 94% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         34 . The method of  claim 20 , wherein said enzyme comprises an amino acid sequence that has at least 95% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         35 . The method of  claim 20 , wherein said enzyme comprises an amino acid sequence that has at least 98% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         36 . The method of  claim 20 , wherein said enzyme comprises an amino acid sequence that has at least 99% sequence identity to any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         37 . The method of  claim 20 , wherein the enzyme comprises the amino acid sequence of any one of SEQ ID NOs: 82, 83, 84, 130, 134, 136, and 138. 
     
     
         38 . The method of  claim 20 , wherein said biocatalyst is a host cell comprising said enzyme. 
     
     
         39 . The method of  claim 38 , wherein said host cell is a microorganism. 
     
     
         40 . The method of  claim 39 , wherein said microorganism is a bacterium, yeast or fungi. 
     
     
         41 . The method of  claim 39 , wherein said microorganism is selected from the group of  Escherichia coli, Bacillus subtilis, Bacillus amyloliquefaciens, Corynebacterium glutamicum, Aspergillus niger, Penicillium chrysogenum, Pichia pastoris , and  Saccharomyces cerevisiae.    
     
     
         42 . The method of claim  1 , wherein said 5-formylpentanoate is prepared by converting an adipate ester or an adipate thioester into said 5-formylpentanoate by an enzyme selected from the group of oxidoreductases. 
     
     
         43 . The method of  claim 42 , wherein the oxidoreductases are selected from the group of aldehyde dehydrogenases (acetylating), fatty acyl-CoA reductases, long-chain-fatty-acyl-CoA reductases, butanal dehydrogenases and succinate semialdehyde dehydrogenases (acetylating). 
     
     
         44 . The method of  claim 42 , wherein the adipate ester or adipate thioester is prepared by converting 2,3-dehydroadipate ester or 2,3-dehydroadipate thioester into the adipate ester or adipate thioester, respectively, by an oxidoreductase acting on the HC—CH group of donors. 
     
     
         45 . The method of  claim 44 , wherein the oxidoreductase acting on the HC—CH group of donors is selected from the group of enoyl-CoA reductases, enoyl-[acyl-carrier-protein]reductases, butyryl-CoA dehydrogenase, acyl-CoA dehydrogenase, and long-chain-acyl-CoA dehydrogenase. 
     
     
         46 . The method of  claim 44 , wherein said 2,3-dehydroadipate ester or 2,3-dehydroadipate thioester is prepared by converting a 3-hydroxyadipate ester or 3-hydroxyadipate thioester to 2,3-dehydroadipate ester or 2,3-dehydroxyadipate thioester, respectively. 
     
     
         47 . The method of  claim 46 , wherein said 3-hydroxyadipate ester or 3-hydroxyadipate thioester is biocatalytically converted in the presence of a biocatalyst comprising an enzyme capable of catalysing the dehydration of a 3-hydroxyacyl ester or 3-hydroxyacyl thioester to 2-enoyl ester or 2-enoyl thioester. 
     
     
         48 . The method of  claim 47 , wherein said enzyme capable of catalysing the dehydration of a 3-hydroxyacyl ester or 3-hydroxyacyl thioester to 2-enoyl ester or 2-enoyl thioester is selected from the group of an enoyl-CoA hydratase, a 3-hydroxybutyryl-CoA dehydratase and a long-chain-enoyl-CoA hydratase. 
     
     
         49 . The method of  claim 46 , wherein said 3-hydroxyadipate ester or 3-hydroxyadipate thioester is prepared by converting a 3-oxoadipate ester or 3-oxoadipate thioester to said 3-hydroxyadipate ester or 3-hydroxyadipate thioester, respectively. 
     
     
         50 . The method of  claim 49 , wherein said 3-oxoadipate ester or 3-oxoadipate thioester is biocatalytically converted in the presence of a biocatalyst comprising an enzyme capable of catalysing the reduction of a carbonyl group to an alcohol group or capable of catalysing the reduction of a 3-oxoacyl ester or 3-oxoacyl thioester to 3-hydroxyacyl ester or 3-hydroxyacyl thioester. 
     
     
         51 . The method of  claim 50 , wherein said enzyme capable of catalysing the reduction of a carbonyl group to an alcohol group or capable of catalysing the reduction of a 3-oxoacyl ester or 3-oxoacyl thioester to 3-hydroxyacyl ester or 3-hydroxyacyl thioester is selected from the group of a 3-hydroxyacyl-CoA dehydrogenase, a 3-hydroxybutanoyl-CoA dehydrogenase, a 3-hydroxypimeloyl-CoA dehydrogenase and a long-chain-3-hydroxyacyl-CoA dehydrogenases. 
     
     
         52 . The method of  claim 46 , wherein said 3-oxoadipate ester or 3-oxoadipate thioester is prepared by converting a succinate ester or succinate thioester and an acetate ester or acetate thioester to 3-oxoadipate ester or 3-oxoadipate thioester. 
     
     
         53 . The method of  claim 52 , wherein said 3-oxoadipate ester or 3-oxoadipate thioester is biocatalytically converted in the presence of a biocatalyst comprising an enzyme capable of acetyl-group transfer. 
     
     
         54 . The method of  claim 53 , wherein said enzyme capable of acetyl-group transfer is selected from the group of an acetyl-CoA:acetyl-CoA C-acetyltransferase, an acyl-CoA:acetyl-CoA C-acetyltransferase and a succinyl-CoA:acetyl-CoA C-succinyltransferase.